# Patent

## Title
Crystals and structure of domain III of West Nile Virus envelope protein in association with a Fab fragment of a neutralizing antibody

## Abstract

The present invention provides crystalline forms of domain III of the West Nile Virus (“WNV”) envelope protein in complex with a Fab fragment of a neutralizing antibody, methods of obtaining such crystals and high-resolution structures and atomic structure coordinates. The crystals of the invention and the atomic structural information are useful for solving crystal and solution structures of related and unrelated proteins, and for screening for, identifying or designing compounds or antibodies that bind to, modulate a biological activity of, or neutralize infection mediated by flaviviral envelope proteins.

## Background
This application is entitled to and claims benefit of U.S. Provisional Application No. 60/619,273, filed Oct. 14, 2004, and of U.S. Provisional Application No. 60/702,803, filed Jul. 26, 2005, each of which is hereby incorporated by reference in its entirety.


The present invention was made, in part, with support from a grant from the United States Government, NIH grant number A161373. The government of the United States may have certain rights in this invention.


1. FIELD OF THE INVENTION
The present invention provides crystalline forms of domain III of the West Nile Virus (“WNV”) envelope protein in complex with a Fab fragment of a neutralizing antibody, methods of obtaining such crystals and high-resolution structures and atomic structure coordinates. The crystals of the invention and the atomic structural information are useful for solving crystal and solution structures of related and unrelated proteins, and for screening for, identifying or designing compounds or antibodies that bind to, modulate a biological activity of, or neutralize infection mediated by flaviviral envelope proteins.
2. BACKGROUND OF THE INVENTION
A member of the flavivirus genus of the Flaviviridae family, WNV is a neurotropic enveloped virus with a single-stranded, positive-polarity 11-kilobase RNA genome and is closely related to viruses that cause dengue fever, yellow fever, and the Japanese, Saint Louis, and Murray Valley encephalitides. WNV cycles between mosquitoes and birds but also infects humans, horses, and other vertebrate species. It is endemic in parts of Africa, Europe, the Middle East, and Asia, and outbreaks throughout the United States during the past five years indicate that it has established its presence in the Western Hemisphere. Infected humans develop a febrile illness that can progress rapidly to a meningitis or fatal encephalitis syndrome. See Granwehr et al., 2004, Lancet Infect Dis 4:547-56; Hubalek et al., 1999, Emerg Inf Dis 5:643-650; and Petersen et al., 2003, JAMA 290:524-8. At present, treatment is supportive and no vaccine exists for human use.
The molecular and structural basis of antibody-mediated protection against WNV and other flaviviruses remains speculative. Based on the sequencing of in vitro neutralization escape variants and the site-specific substitution of specific charged or polar residues, most neutralizing antibodies against flaviviruses appear to localize to domain III. See Beasley and Aaskov, 2001, Virology 279:447-58; Beasley et al., 2002, J. Virol. 76:13097-13100; Cecilia and Gould, 1991, Virology 181:70-7; Crill and Roehrig, 2001, J. Virol. 75:7769-73; Lin et al., 1994, Virology 202:885-90; Roehrig et al., 1983, Virology 128:118-26; Schlesinger et al., 1996, J. Gen. Virol. 77:1277-85; Seifet al., 1995, Vaccine 13:1515-21; Volk et al., 2004, J. Biol. Chem. 279:38755-38761; and Wu et al., 1997, Virus. Res. 51:173-81. However, putative contact residues for individual mAbs that have been identified by neutralization escape may be flawed because mutations may cause local unfolding that abolish multiple antibody epitopes. Moreover, many of these studies did not confirm that the mapped neutralizing mAbs also abolished infection in vivo in animals. As an alternative strategy, one group recently used NMR to map a neutralizing mAb against Japanese encephalitis virus (JEV). See Wu et al., 2003, J. Biol. Chem. 278:46007-46013. Chemical shifts in domain III of JEV were detected after mAb binding at residues E302-312, E322-329, E360-372, and E385-392, corresponding to the top portion of an exposed β-barrel. Although additional information was obtained, NMR and its solution structure do not provide any insight as to the structural basis for antibody recognition of the neutralizing epitope.
Recently, prophylactic and therapeutic efficacy of pooled, immune human γ-globulin has been demonstrated in mice infected with WNV. See Agrawal and Petersen, 2003, J. Infect. Dis. 188:1-4; Ben-Nathan et al., 2003, J. Infect. Dis. 188:5-12; and Engle et al., 2003, J. Virol. 77:12941-12949. Because human γ-globulin is made from human blood plasma, it has an inherent risk of transmitting known and unknown infectious agents. More recently, a monoclonal antibody (mAb) therapeutic against WNV E protein has been developed that is  ̃1,000-fold more potent that pooled human γ-globulin in its ability to neutralize virus infection in vitro and in vivo. This antibody (E16), which recognizes domain III, was cloned, humanized, expressed and confirmed as therapeutically effective in an established mouse model of WNV infection.
Nonetheless, additional antibodies that can bind and neutralize WNV more strongly may be needed for effective treatment and/or prevention of WNV infection. Such antibodies could be designed and constructed by identifying amino acids of the antibodies that mediate the antigen-antibody interaction. These amino acids can be selectively altered to generate antibody variants that could be screened for enhanced WNV binding and/or neutralization.
Further, small molecule therapeutics that can mimic antibodies that bind and/or neutralize WNV infection would also be of use, as such small molecules may be easier and less expensive to manufacture and easier to administer orally. Such small molecule therapeutics could be designed based on the three-dimensional structure coordinates of an antibody that binds domain III of WNV E protein in complex with the domain III.
In addition, a small molecule therapeutic such as an antigen that mimics the domain III epitope recognized by E16 could be administered to generate an immune response against WNV. A composition comprising an antigen that mimics WNV would provide a safer method of preventing WNV infection. An effective antigen mimic of WNV could be administered, to persons with a functioning immune system, as an immunoprophylactic to raise an immune response against the virus with minimal or no danger of infection caused by the immunoprophylactic itself.
Still further, a small molecule therapeutic that can interact with antibody-WNV complexes to stimulate antibody-mediated neutralization of WNV infection would also be of significant utility in treatment and/or prevention of WNV infection. In addition, a greater understanding of the interaction between neutralizing antibodies and domain III of WNV E protein is needed to inform strategies for designing vaccines that will elicit strong and broadly neutralizing immune responses. The present invention provides a substantial advancement towards these and other unrealized needs.
3. SUMMARY OF THE INVENTION
In one aspect, the invention provides crystalline forms of polypeptide complexes corresponding to a Fab fragment of a neutralizing antibody in complex with domain III of the WNV E protein. The crystals of the invention comprise crystallized polypeptide complexes corresponding to the wild-type or mutated domain III of WNV E protein in complex with a Fab fragment of a neutralizing antibody, e.g., a Fab fragment prepared from monoclonal antibody E16 (WNV DIII-E16 Fab complex). The crystals of the invention include native crystals, in which the crystallized WNV DIII-E16 Fab complex is substantially pure; heavy-atom atom derivative crystals, in which the crystallized WNV DIII-E16 Fab complex is in association with one or more heavy-metal atoms; and poly-crystals, in which the crystallized WNV DIII-E16 Fab complex is in association with one or more additional compounds, including but not limited to, cofactors, ligands, substrates, substrate analogs, inhibitors, allosteric effectors, etc. to form a crystalline poly-complex. Preferably, such compounds bind a catalytic or active site. The poly-crystals may be native poly-crystals, in which the poly-complex is substantially pure, or they may be heavy-atom derivative poly-crystals, in which the poly-complex is in association with one or more heavy-metal atoms.
In certain embodiments, the crystals of the invention are generally characterized by a unit cell of a=52.4+/−0.2 Å, b=83.3+/−0.2 Å, c=110.6+/−0.2 Å, and are preferably of diffraction quality. A typical diffraction pattern is illustrated in FIG. 4. In more preferred embodiments, the crystals of the invention are of sufficient quality to permit the determination of the three-dimensional X-ray diffraction structure of the crystalline polypeptide(s) to high resolution, preferably to a resolution of greater than about 3 Å, typically in the range of about 2 Å to about 3 Å.
The invention also provides methods of making the crystals of the invention. Generally, crystals of the invention are grown by dissolving substantially pure polypeptide complexes in an aqueous buffer that includes a precipitant at a concentration just below that necessary to precipitate the polypeptide complexes. Water is then removed by controlled evaporation to produce precipitating conditions, which are maintained until crystal growth ceases.
Poly-crystals of the invention are prepared by soaking a native crystal prepared according to the above method in a liquor comprising the compound to be added to the poly-crystal of the desired complex. Alternatively, the poly-crystals may be prepared by co-crystallizing the polypeptide complexes in the presence of the compound according to the method discussed above.
Heavy-atom derivative crystals of the invention may be prepared by soaking native crystals or poly-crystals prepared according to the above method in a liquor comprising a salt of a heavy atom or an organometallic compound. Alternatively, heavy-atom derivative crystals may be prepared by crystallizing a polypeptide complex comprising selenomethionine and/or selenocysteine residues according to the methods described previously for preparing native crystals.
In another aspect, the invention provides machine-computer-readable media embedded with the three-dimensional structural information obtained from the crystals of the invention, or portions or substrates thereof. Such three-dimensional structural information will typically include the atomic structure coordinates of the crystallized polypeptide complex or poly-complex, or the atomic structure coordinates of a portion thereof such as, for example, an active or binding site, but may include other structural information, such as vector representations of the atomic structures coordinates, etc. The types of machine- or computer-readable media into which the structural information is embedded typically include magnetic tape, floppy discs, hard disc storage media, optical discs, CD-ROM, DVD, electrical storage media such as RAM or ROM, and hybrids of any of these storage media. Such media further include paper on which is recorded the structural information that can be read by a scanning device and converted into a three-dimensional structure with an OCR. The machine-readable media of the invention may further comprise additional information that is useful for representing the three-dimensional structure, including, but not limited to, thermal parameters, chain identifiers, and connectivity information.
The invention is illustrated by way of working examples demonstrating the crystallization and characterization of crystals, the collection of diffraction data, and the determination and analysis of the three-dimensional structure of WNV DIII-E16 Fab complex.
The atomic structure coordinates and machine-readable media of the invention have a variety of uses. For example, the coordinates are useful for solving the three-dimensional X-ray diffraction and/or solution structures of other proteins, including mutant WNV E proteins, both alone or in complex with E16 Fab, or a portion thereof, complexes comprising WNV E protein and E16 Fab in combination with one or more additional compounds, and unrelated proteins, to high resolution. Structural information may also be used in a variety of molecular modeling and computer-based screening applications to, for example, intelligently design mutants of the WNV E protein and/or a neutralizing antibody, or fragments thereof, that have altered biological activity, to identify additional neutralizing antibodies, or fragments thereof, that bind WNV DIII, to identify antibodies that bind neutralizing epitopes of WNV DIII in preference to enhancing epitopes of WNV DIII, to intelligently design antibodies, or fragments thereof, that bind WNV III with high affinity, or to design vaccines capable of inducing immune responses that inhibit WNV E protein binding.
Thus, in other aspects, the invention provides methods that comprise mutating an amino acid of the antibody which when mutated is predicted to increase the affinity with which the antibody binds the domain III of WNV E protein.
In still other aspects, the invention provides a method for identifying a neutralizing epitope of a domain III of a flaviviral envelope protein, comprising comparing an amino acid sequence of the domain III of the flaviviral envelope protein to an amino acid sequence of a domain III of a WNV E protein, and identifying the amino acids of the domain III of the flaviviral envelope protein corresponding to a neutralizing epitope of the domain III of the WNV E protein, thereby identifying the neutralizing epitope of the flaviviral envelope protein.
In yet other aspects, the invention provides a purified antibody that specifically binds a neutralizing epitope of domain III of WNV E protein, with the proviso that the antibody is not 5H10, 3A3, 5C5, 7H2, 11C, 17C8, 10C5, 8B10, E1, E16, E24, E27, E33, E34, E40, E43, E47, E49, and/or E58. These monoclonal antibodies are described in Beasley & Barrett, 2002, J. Virol 76:13097-13100; Oliphant et al., 2005, Nature Medicine 11:522-530; and Sanchez et al., 2005, Virology 336:70-82, each of which is hereby incorporated by reference in its entirety.
In still other aspects, the invention provides a pharmaceutical composition comprising an antibody of the invention and a pharmaceutically acceptable excipient, diluent, vehicle, or carrier.
In yet other aspects, the invention provides a kit comprising a pharmaceutical composition of the invention.
In still other aspects, the invention provides a method of treating WNV infection, comprising administering to a subject an effective amount of a purified antibody that binds a neutralizing epitope of domain III of WNV E protein, with the proviso that the antibody is not 5H10, 3A3, 5C5, 7H2, 11C, 17C8, 10C5, 8B10, E1, E16, E24, E27, E33, E34, E40, E43, E47, E49, and/or E58.
In yet other aspects, the invention provides a method of preventing WNV infection, comprising administering to a subject an effective amount of a purified antibody that binds a neutralizing epitope of domain III of WNV E protein with higher affinity than the antibody binds an enhancing epitope of domain III of WNV E protein at neutralizing concentrations, with the proviso that the antibody is not 5H10, 3A3, 5C5, 7H2, 11C, 17C8, 10C5, 8B10, E1, E16, E24, E27, E33, E34, E40, E43, E47, E49, and/or E58.
In still other aspects, the invention provides a WNV immunogen comprising a peptide corresponding to amino acids 300-309, 330-333, 365-368, or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the peptide is arranged in the three dimensional conformation of amino acids 300-309, 330-333, 365-368, or 389-391, respectively, of WNV E protein according to the structural coordinates shown in Table 7. In certain embodiments, the immunogen does not comprise the entire amino acid sequence of domain III of WNV E protein.
In yet other aspects, the invention provides a pharmaceutical composition comprising a WNV immunogen of the invention and a pharmaceutically acceptable excipient, diluent, vehicle, or carrier.
In still other aspects, the invention provides a method of inducing in a subject an immune response against domain III of WNV E protein, comprising administering to a subject a WNV immunogen of the invention.
The atomic structure coordinates and machine-readable media may also be used to computationally design and to identify compounds that bind the polypeptide(s) or a portion or fragment of the polypeptide(s), such as the active site. Such compounds may be used as lead compounds in pharmaceutical efforts to identify compounds as a therapeutic or prophylactic approach toward the prevention or treatment of, e.g., WNV or other flaviviral infection.
The examples demonstrate that the crystal structure of WNV DIII-E16 Fab complex has been determined to 2.5 Å resolution.
3.1 Abbreviations
The amino acid notations used herein for the twenty genetically encoded L-amino acids are conventional and are as follows:
















One-Letter
Three-Letter



Amino Acid
Symbol
Symbol









Alanine
A
Ala



Arginine
R
Arg



Asparagine
N
Asn



Aspartic acid
D
Asp



Cysteine
C
Cys



Glutamine
Q
Gln



Glutamic acid
E
Glu



Glycine
G
Gly



Histidine
H
His



Isoleucine
I
Ile



Leucine
L
Leu



Lysine
K
Lys



Methionine
M
Met



Phenylalanine
F
Phe



Proline
P
Pro



Serine
S
Ser



Threonine
T
Thr



Tryptophan
W
Trp



Tyrosine
Y
Tyr



Valine
V
Val










As used herein, unless specifically delineated otherwise, the three-letter amino acid abbreviations designate amino acids in the L-configuration. Amino acids in the D-configuration are preceded with a “D-.” For example, Arg designates L-arginine and D-Arg designates D-arginine. Likewise, the capital one-letter abbreviations refer to amino acids in the L-configuration. Lower-case one-letter abbreviations designate amino acids in the D-configuration. For example, “R” designates L-arginine and “r” designates D-arginine.
Unless noted otherwise, when polypeptide sequences are presented as a series of one-letter and/or three-letter abbreviations, the sequences are presented in the N→C direction, in accordance with common practice.
The abbreviation “WNV” is used herein to refer to “West Nile Virus.”
The abbreviation “E protein” is used herein to refer to “envelope protein.”
The abbreviation “DIII” is used herein to refer to “domain III of envelope protein.”
3.2 DEFINITIONS
As used herein, the following terms shall have the following meanings:
“Genetically Encoded Amino Acid” refers to L-isomers of the twenty amino acids that are defined by genetic codons. The genetically encoded amino acids are the L-isomers of glycine, alanine, valine, leucine, isoleucine, serine, methionine, threonine, phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine, aspartic acid, asparagine, glutamic acid, glutamine, arginine and lysine.
“Genetically Non-Encoded Amino Acid” refers to amino acids that are not defined by genetic codons. Genetically non-encoded amino acids include derivatives or analogs of the genetically-encoded amino acids that are capable of being enzymatically incorporated into nascent polypeptides using conventional expression systems, such as selenomethionine (SeMet) and selenocysteine (SeCys); isomers of the genetically-encoded amino acids that are not capable of being enzymatically incorporated into nascent polypeptides using conventional expression systems, such as D-isomers of the genetically-encoded amino acids; L- and D-isomers of naturally occurring a-amino acids that are not defined by genetic codons, such as α-aminoisobutyric acid (Aib); L- and D-isomers of synthetic α-amino acids that are not defined by genetic codons, such as ______; and other amino acids such as β-amino acids, γ-amino acids, etc. In addition to the D-isomers of the genetically-encoded amino acids, common genetically non-encoded amino acids include, but are not limited to norleucine (Nle), penicillamine (Pen), N-methylvaline (MeVal), homocysteine (hCys), homoserine (hSer), 2,3-diaminobutyric acid (Dab) and omithine (Orn). Additional exemplary genetically non-encoded amino acids are found, for example, in Practical Handbook of Biochemistry and Molecular Biology, 1989, Fasman, Ed., CRC Press, Inc., Boca Raton, Fla., pp. 3-76 and the various references cited therein.
“Hydrophilic Amino Acid” refers to an amino acid having a side chain exhibiting a hydrophobicity of less than zero according to the normalized consensus hydrophobicity scale of Eisenberg et al., 1984, J. Mol. Biol. 179:125-142. Genetically encoded hydrophilic amino acids include Thr (T), Ser (S), His (H), Glu (E), Asn (N), Gln (Q), Asp (D), Lys (K) and Arg (R). Genetically non-encoded hydrophilic amino acids include the D-isomers of the above-listed genetically-encoded amino acids, ornithine (Orn), 2,3-diaminobutyric acid (Dab) and homoserine (hSer).
“Acidic Amino Acid” refers to a hydrophilic amino acid having a side chain pK value of less than 7 under physiological conditions. Acidic amino acids typically have negatively charged side chains at physiological pH due to loss of a hydrogen ion. Genetically encoded acidic amino acids include Glu (E) and Asp (D). Genetically non-encoded acidic amino acids include D-Glu (e) and D-Asp (d).
“Basic Amino Acid” refers to a hydrophilic amino acid having a side chain pK value of greater than 7 under physiological conditions. Basic amino acids typically have positively charged side chains at physiological pH due to association with hydronium ion. Genetically encoded basic amino acids include His (H), Arg (R) and Lys (K). Genetically non-encoded basic amino acids include the D-isomers of the above-listed genetically-encoded amino acids, ornithine (Orn) and 2,3-diaminobutyric acid (Dab).
“Polar Amino Acid” refers to a hydrophilic amino acid having a side chain that is uncharged at physiological pH, but which comprises at least one covalent bond in which the pair of electrons shared in common by two atoms is held more closely by one of the atoms. Genetically encoded polar amino acids include Asn (N), Gln (Q), Ser (S), and Thr (T). Genetically non-encoded polar amino acids include the D-isomers of the above-listed genetically-encoded amino acids and homoserine (hSer).
“Hydrophobic Amino Acid” refers to an amino acid having a side chain exhibiting a hydrophobicity of greater than zero according to the normalized consensus hydrophobicity scale of Eisenberg et al., 1984, J. Mol. Biol. 179:125-142. Genetically encoded hydrophobic amino acids include Pro (P), Ile (I), Phe (F), Val (V), Leu (L), Trp (W), Met (M), Ala (A), Gly (G) and Tyr (Y). Genetically non-encoded hydrophobic amino acids include the D-isomers of the above-listed genetically-encoded amino acids, norleucine (Nle) and N-methyl valine (MeVal).
“Aromatic Amino Acid” refers to a hydrophobic amino acid having a side chain comprising at least one aromatic or heteroaromatic ring. The aromatic or heteroaromatic ring may contain one or more substituents such as —OH, —SH, —CN, —F, —Cl, —Br, —I, —NO2, —NO, —NH2, —NHR, —NRR, —C(O)R, —C(O)OH, —C(O)OR, —C(O)NH2, —C(O)NHR, —C(O)NRR and the like where each R is independently (C1-C6) alkyl, (C1-C6) alkenyl, or (C1-C6) alkynyl. Genetically encoded aromatic amino acids include Phe (F), Tyr (Y), Trp (W) and His (H). Genetically non-encoded aromatic amino acids include the D-isomers of the above-listed genetically-encoded amino acids.
“Apolar Amino Acid” refers to a hydrophobic amino acid having a side chain that is uncharged at physiological pH and which has bonds in which the pair of electrons shared in common by two atoms is generally held equally by each of the two atoms (i.e., the side chain is not polar). Genetically encoded apolar amino acids include Leu (L), Val (V), Ile (I), Met (M), Gly (G) and Ala (A). Genetically non-encoded apolar amino acids include the D-isomers of the above-listed genetically-encoded amino acids, norleucine (Nle) and N-methyl valine (MeVal).
“Aliphatic Amino Acid” refers to a hydrophobic amino acid having an aliphatic hydrocarbon side chain. Genetically encoded aliphatic amino acids include Ala (A), Val (V), Leu (L) and Ile (I). Genetically non-encoded aliphatic amino acids include the D-isomers of the above-listed genetically-encoded amino acids, norleucine (Nle) and N-methyl valine (MeVal).
“Helix-Breaking Amino Acid” refers to those amino acids that have a propensity to disrupt the structure of α-helices when contained at internal positions within the helix. Amino acid residues exhibiting helix-breaking properties are well-known in the art (see, e.g., Chou & Fasman, 1978, Ann. Rev. Biochem. 47:251-276) and include Pro (P), D-Pro (p), Gly (G) and potentially all D-amino acids (when contained in an L-polypeptide; conversely, L-amino acids disrupt helical structure when contained in a D-polypeptide).
“Cysteine-like Amino Acid” refers to an amino acid having a side chain capable of participating in a disulfide linkage. Thus, cysteine-like amino acids generally have a side chain containing at least one thiol (—SH) group. Cysteine-like amino acids are unusual in that they can form disulfide bridges with other cysteine-like amino acids. The ability of Cys (C) residues and other cysteine-like amino acids to exist in a polypeptide in either the reduced free —SH or oxidized disulfide-bridged form affects whether they contribute net hydrophobic or hydrophilic character to a polypeptide. Thus, while Cys (C) exhibits a hydrophobicity of 0.29 according to the consensus scale of Eisenberg (Eisenberg, 1984, supra), it is to be understood that for purposes of the present invention Cys (C) is categorized as a polar hydrophilic amino acid, notwithstanding the general classifications defined above. Other cysteine-like amino acids are similarly categorized as polar hydrophilic amino acids. Typical cysteine-like residues include, for example, penicillamine (Pen), homocysteine (hCys), etc.
As will be appreciated by those of skill in the art, the above-defined classes or categories are not mutually exclusive. Thus, amino acids having side chains exhibiting two or more physico-chemical properties can be included in multiple categories. For example, amino acid side chains having aromatic groups that are further substituted with polar substituents, such as Tyr (Y), may exhibit both aromatic hydrophobic properties and polar or hydrophilic properties, and could therefore be included in both the aromatic and polar categories. Typically, amino acids will be categorized in the class or classes that most closely define their net physico-chemical properties. The appropriate categorization of any amino acid will be apparent to those of skill in the art.
The classifications of the genetically encoded and common non-encoded amino acids according to the categories defined above are summarized in Table I, below. It is to be understood that Table I is for illustrative purposes only and does not purport to be an exhaustive list of the amino acid residues belonging to each class. Other amino acid residues not specifically mentioned herein can be readily categorized based on their observed physical and chemical properties in light of the definitions provided herein.







TABLE I







CLASSIFICATIONS OF COMMONLY


ENCOUNTERED AMINO ACIDS










Genetically
Genetically


Classification
Encoded
Non-Encoded





Hydrophobic




Aromatic
F, Y, W, H
f, y, w, h


Apolar
L, V, I, M, G, A, P
l, v, i, m, a, p, Nle, MeVal


Aliphatic
A, V, L, I
a, v, l, I, Nle, MeVal


Hydrophilic


Acidic
D, E
d, e


Basic
H, K, R
h, k, r, Orn, Dab


Polar
C, Q, N, S, T
c, q, n, s, t, hSer


Helix-Breaking
P, G
P









“Wild-type domain III of West Nile Virus E protein Domain III” or “wild-type domain III of WNV E protein” refers to a polypeptide having an amino acid sequence that corresponds identically to the amino acid sequence of a naturally-occurring domain III of WNV E protein.
A “neutralizing antibody” refers to a monoclonal antibody that can bind to an envelope protein of a flavivirus and reduce or prevent infection by a flavivirus of a permissive cell.
An “antibody” or “antibodies” refers to monoclonal antibodies, multispecific antibodies, human antibodies, humanized antibodies, synthetic antibodies, chimeric antibodies, camelized antibodies, single-chain Fvs (scFv), single chain antibodies, Fab fragments, F(ab′) fragments, disulfide-linked Fvs (sdFv), intrabodies, and anti-idiotypic (anti-Id) antibodies (including, e.g., anti-Id and anti-anti-Id antibodies), bispecific, and epitope-binding fragments of any of the above. In particular, antibodies include immunoglobulin molecules and immunologically active fragments of immunoglobulin molecules, i.e., molecules that contain an antigen binding site. Immunoglobulin molecules can be of any type (e.g., IgG, IgE, IgM, IgD, IgA and IgY), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1 and IgA2) or subclass.
A “Fab” or a “Fab fragment” refers to a portion of a monoclonal antibody that corresponds to the constant and variable regions of an immunoglobulin light chain in association with the first constant region (CH1 domain), variable region, and a portion of the hinge region of an immunoglobulin heavy chain.
“Domain III of West Nile Virus E protein comiplexed with E16 Fab,” “Domain III of WNV E protein complexed with E16 Fab,” or “WNV DIII-E16 Fab complex,” or “WNV DIII-Fab complex” refers to E16 Fab in association with domain III of WNV E protein, as each of those terms is defined herein.
“Crystallized domain III of West Nile Virus E Protein complexed with a Fab fragment of a neutralizing antibody,” “crystallized domain III of WNV E protein complexed with a Fab fragment of a neutralizing antibody,” “crystallized WNV DIII-Fab complex,” or “crystallized WNV DIII-E16 Fab complex” refers to a complex comprising a polypeptide having an amino acid sequence which corresponds identically to SEQ ID NO:1, a polypeptide having an amino acid sequence which corresponds exactly to SEQ ID NO:2, and a polypeptide having an amino acid sequence which corresponds exactly to SEQ ID NO:3, and which complex is in crystalline form. Thus, “crystallized domain III of West Nile Virus E Protein complexed with a Fab fragment of a neutralizing antibody,” “crystallized domain III of WNV E protein complexed with a Fab fragment of a neutralizing antibody,” or “crystallized WNV DIII-Fab complex” each refers to a complex of polypeptides comprising domain III of WNV E protein complexed with a Fab fragment of the neutralizing antibody E16.
“Association” refers to a condition of proximity between a chemical entity or compound, or portions or fragments thereof, and a polypeptide, or portions or fragments thereof. The association may be non-covalent, i.e., where the juxtaposition is energetically favored by, e.g., hydrogen-bonding, van der Waals, electrostatic or hydrophobic interactions, or it may be covalent.
“Poly-Complex,” as used herein, refers to a polypeptide complex in association with one or more additional compounds beyond those present in the polypeptide complex. Such compounds include, by way of example and not limitation, cofactors, ligands, substrates, substrate analogues, inhibitors, allosteric effectors, etc.
“Domain III” of WNV E protein or “WNV DIII” refers to amino acids 296 to 415 of WNV E protein.
“Mutant” refers to a polypeptide or combination of polypeptides characterized by an amino acid sequence that differs from the wild-type sequence(s) by the substitution of at least one amino acid residue of the wild-type sequence(s) with a different amino acid residue and/or by the addition and/or deletion of one or more amino acid residues to or from the wild-type sequence(s). The additions and/or deletions can be from an internal region of the wild-type sequence and/or at either or both of the N- or C-termini. A mutant may have, but need not have, receptor binding, in the case of mutant WNV E proteins, or neutralization, in the case of mutant antibodies or antibody fragments, activity. Preferably, a mutant displays biological activity that is substantially similar to that of the wild-type WNV E protein or antibody or antibody fragment.
“Conservative Mutant” refers to a mutant in which at least one amino acid residue from the wild-type sequence(s) is substituted with a different amino acid residue that has similar physical and chemical properties, i.e., an amino acid residue that is a member of the same class or category, as defined above. For example, a conservative mutant may be a polypeptide or combination of polypeptides that differs in amino acid sequence from the wild-type sequence(s) by the substitution of a specific aromatic Phe (F) residue with an aromatic Tyr (Y) or Trp (W) residue.
“Non-Conservative Mutant” refers to a mutant in which at least one amino acid residue from the wild-type sequence(s) is substituted with a different amino acid residue that has dissimilar physical and/or chemical properties, i.e., an amino acid residue that is a member of a different class or category, as defined above. For example, a non-conservative mutant may be a polypeptide or combination of polypeptides that differs in amino acid sequence from the wild-type sequence by the substitution of an acidic Glu (E) residue with a basic Arg (R), Lys (K) or Orn residue.
“Deletion Mutant” refers to a mutant having an amino acid sequence or sequences that differs from the wild-type sequence(s) by the deletion of one or more amino acid residues from the wild-type sequence(s). The residues may be deleted from internal regions of the wild-type sequence(s) and/or from one or both termini.
“Truncated Mutant” refers to a deletion mutant in which the deleted residues are from the N- and/or C-terminus of the wild-type sequence(s).
“Extended Mutant” refers to a mutant in which additional residues are added to the N- and/or C-terminus of the wild-type sequence(s).
“Methionine mutant” refers to (1) a mutant in which at least one methionine residue of the wild-type sequence(s) is replaced with another residue, preferably with an aliphatic residue, most preferably with a Leu (L) or Ile (I) residue; or (2) a mutant in which a non-methionine residue, preferably an aliphatic residue, most preferably a Leu (L) or Ile (I) residue, of the wild-type sequence(s) is replaced with a methionine residue.
“Selenomethionine mutant” refers to (1) a mutant which includes at least one selenomethionine (SeMet) residue, typically by substitution of a Met residue of the wild-type sequence(s) with a SeMet residue, or by addition of one or more SeMet residues at one or both termini, or (2) a methionine mutant in which at least one Met residue is substituted with a SeMet residue. Preferred SeMet mutants are those in which each Met residue is substituted with a SeMet residue.
“Cysteine mutant” refers to (1) a mutant in which at least one cysteine residue of the wild-type sequence(s) is replaced with another residue, preferably with a Ser (S) residue; or (2) a mutant in which a non-cysteine residue, preferably a Ser (S) residue, of the wild-type sequence(s) is replaced with a cysteine residue.
“Selenocysteine mutant” refers to (1) a mutant which includes at least one selenocysteine (SeCys) residue, typically by substitution of a Cys residue of the wild-type sequence(s) with a SeCys residue, or by addition of one or more SeCys residues at one or both termini, or (2) a cysteine mutant in which at least one Cys residue is substituted with a SeCys residue. Preferred SeCys mutants are those in which each Cys residue is substituted with a SeCys residue.
“Homologue” refers to a polypeptide having at least 80% amino acid sequence identity or having a BLAST score of 1×10−6 over at least 100 amino acids (Altschul et al., 1997, Nucleic Acids Res. 25:3389-402) with WNV E protein or any functional domain, e.g., domain III, of WNV E protein, as defined by Mukhopadhyay et al., 2003, Science 302:248.
“Polypeptide complex” refers to an association of two or more, usually three polypeptides. For example, a polypeptide complex as used herein includes domain III of WNV E protein in association with a Fab fragment of a neutralizing antibody, as each of these terms is defined herein.
“Crystal” refers to a composition comprising a polypeptide complex in crystalline form. The term “crystal” includes native crystals, heavy-atom derivative crystals and poly-crystals, as defined herein.
“Native Crystal” refers to a crystal wherein the polypeptide complex is substantially pure. As used herein, native crystals do not include crystals of polypeptide complexes comprising amino acids that are modified with heavy atoms, such as crystals of selenomethionine mutants, selenocysteine mutants, etc.
“Heavy-atom Derivative Crystal” refers to a crystal wherein the polypeptide complex is in association with one or more heavy-metal atoms. As used herein, heavy-atom derivative crystals include native crystals into which a heavy metal atom is soaked, as well as crystals of selenomethionine mutants and selenocysteine mutants.
“Poly-Crystal” refers to a composition comprising a poly-complex, as defined above, in crystalline form. Poly-crystals include native poly-crystals and heavy-atom derivative poly-crystals.
“X-ray Diffraction” refers to a type of wave interference created when high energy X-ray radiation interacts with any obstruction in its traveling path. The obstruction is often in the form of a crystal of protein, nucleic acid, or inorganic compound. The electrons that surround the atoms in the crystal, rather than the atomic nuclei, are the entities which physically interact with the incoming X-ray photons. When X-ray radiation hits the atoms in a crystal, they make the electronic clouds of the atoms move as does any electromagnetic wave. The re-emitted X-ray radiation gives rise to constructive or destructive interferences. This phenomenon is called X-ray diffraction. In X-ray crystallography, the X-ray diffraction patterns of closely spaced lattices of atoms in the crystal are recorded and then analyzed to reveal the structural nature of the crystal. For example, the spacing between the crystal lattices can be determined using Bragg's law. X-ray diffraction is widely used in chemistry and biochemistry to determine the structures of an immense variety of molecules, including inorganic compounds, DNA and proteins. X-ray diffraction is commonly carried out using single crystals of a material, but if these are not available, microcrystalline powdered samples may also be used, although this requires different equipment. A detailed discussion on X-ray diffraction may be found in Chapter 4 in “Principles of Protein X-ray Crystallography” by Drenth, second edition 1999, Springer-Verlag Inc.
“Bragg's Law” refers to the principle that defines the diffraction conditions that give rise to constructive interferences. When the phase shift of the incident radiation is proportional to 2π, the condition can be expressed as: nλ=2d sin(θ), where n is an integer; λ is the wavelength of the X-ray radiation, or radiations caused by moving electrons, protons and neutrons; d is the spacing between the planes in the atomic lattice, and θ is the angle between the incident ray and the scattering planes.
“Laue Diffraction” refers to the diffraction pattern obtained from a stationary crystal exposed to a continuous range of wavelengths of X-rays (e.g. polychromatic or “white” radiation). The application of monochromatic X-ray radiation limited the use of Laue diffraction until the availability of synchrotrons that provide fully polychromatic beams with smooth spectral profiles. Synchrotron radiations have high intensity, very small divergence, which renders them ideal sources for Laue diffraction of protein crystals. There are at least two practical variants of the Laue method, the back-reflection and the transmission Laue diffraction. In the back-reflection method, the reflection recorder is placed between the X-ray source and the crystal. The beams that are diffracted in a backward direction are recorded. One side of the cone of Laue reflections is defined by the transmitted beam. The recorder intersects the cone, with the diffraction spots generally lying on a hyperbola. In the transmission Laue diffraction, the reflection recorder is placed behind the crystal to record beams which are transmitted through the crystal. One side of the cone of Laue reflections is defined by the transmitted beam. The recorder intersects the cone, with the diffraction spots generally lying on an ellipse. Under Laue diffraction, protein diffraction pattern at high intensity synchrotron X-ray sources can be taken in times as short as 150 picoseconds (Srajer et al., 1996, Science 274:1726-1729). The greatest advantage of Laue diffraction is its time efficiency under synchrotron radiations. Laue diffraction is extensively discussed in “Time resolved macromolecular crystallography,” by Cruickshank et al., 1992, Oxford University Press.
“Neutron Diffraction” refers to a crystallography technique that uses neutrons to determine the atomic structure of a material. Neutrons are particles found in the atomic nucleus. In a nuclear reactor, neutrons can be set free when nuclei decay (fission, radioactivity). All quantum particles can exhibit wave phenomena we typically associate with light or sound. Diffraction is one of these phenomena; it occurs when waves encounter obstacles whose size is comparable with the wavelength. If the wavelength of a quantum particle is short enough, atoms or their nuclei can serve as diffraction obstacles. When neutrons from a reactor are slowed down and selected properly, their wavelength lies near one angstrom (0.1 nanometer), the typical separation between atoms in a solid material. A neutron diffraction measurement typically uses a neutron source (e.g. a nuclear reactor or spallation source), a target (the material to be studied), and a detector. Other components may be needed to select the desired neutron wavelength. Some parts of the setup may also be movable. Since neutrons are not charged, they do not interact with the electron cloud surrounding the atom (unlike X-ray or electron diffraction). The neutrons will only interact with the nucleus of the atom. Thus, neutron diffraction reveals the atomic structure but not the charge distribution around the atom, although the two are usually very similar. Neutron diffraction reveals structural details of the target material, which are measured by recording the way in which neutrons are deflected. Neutrons can also change their speed during the scattering experiment; this can be used to study the types of vibrations that can occur in a solid. An important difference between neutron and X-ray diffraction is that neutrons are sensitive to magnetic forces in the material. The application of neutron diffraction in protein structure determination, in particular in determining the hydration level of protein crystals, is discussed in detail in articles by Cheng and Schoenborn, 1990, Acta Cryst. B46: 195-208; Langan et al., 2004, J. Appl. Cryst. 37:24-31; and Steinbach and Brooks, 1993, Proc. Natl. Acad. Sci. USA 90:9135-9139.
“Electron Diffraction” refers the diffractions where the incident radiation is created by fast-moving electrons. The electrons are deflected not as particles but as waves, as in classical diffraction. The technique is typically used on crystal samples that have a regularly spaced atomic lattice. Most electron diffraction is performed with high energy electrons whose wavelengths are orders of magnitude smaller than the interplanar spacing in most crystals. For example, for 100 keV electrons, their wavelength λ will be shorter than 3.7×10−12 m. Typical lattice parameters for crystals are around 0.3 nanometers. The electrons are scattered by interaction with the positively charged atomic nuclei. Electrons are charged particles that interact very strongly with solids, so their penetration of crystals is very limited. Low-energy Electron Diffraction (LEED) and Reflection High-Energy Electron Diffraction (RHEED) are therefore considered to be surface science techniques, while transmission electron diffraction is limited to specimens less than 1 mm thick. In recent studies, however, electron diffraction has been applied to detect structural changes in the photo cycle of bacteriorhodopsin (Subramaniam et al., 1993, EMBO J. 12:1-8).
“Crystallization” in the context of protein X-ray crystallography refers to the processes during which soluble proteins are transformed into their crystalline forms. Crystals of a protein can be grown out of its solution state under experimental conditions that allow controlled phase transition. Such experimental conditions include a mixture of multiple solutions that often contain an aqueous solution of the target protein, a solution of one or a mixture of precipitants, and one or more compounds that contribute to the overall pH or ionic strength of the final mixture.
“Mother liquor” refers to the impure or complex residual solution that remains after the crystallization process. Once crystals are formed, they can be preserved in mother liquor when other experimental conditions remain unchanged. Solutions resembling the composition of a mother liquor are often used as carrier solutions for incorporating additional reagents into the already formed crystals, such as introducing heavy atoms or cryoprotectants.
“Diffraction Quality Crystal” refers to a crystal that is well-ordered and of a sufficient size, i.e., at least 10 μm, preferably at least 50 μm, and most preferably at least 100 μm in its smallest dimension such that it produces measurable diffraction to at least 3 Å resolution, preferably to at least 2 Å resolution, and most preferably to at least 1.5 Å resolution or lower. Diffraction quality crystals include native crystals, heavy-atom derivative crystals, and poly-crystals.
“Unit Cell” refers to the smallest and simplest volume element (i.e., parallelpiped-shaped block) of a crystal that is completely representative of the unit or pattern of the crystal, such that the entire crystal can be generated by translation of the unit cell. The dimensions of the unit cell are defined by six numbers: dimensions a, b and c and angles α, β and γ (Blundel et al., 1976, Protein Crystallography, Academic Press). A crystal is an efficiently packed array of many unit cells.
“Triclinic Unit Cell” refers to a unit cell in which a ≠b≠c and α≠β≠γ.
“Monoclinic Unit Cell” refers to a unit cell in which a≠b≠c; α=γ=90°; and β≠90°, defined to be ≧90°.
“Orthorhombic Unit Cell” refers to a unit cell in which a≠b≠c; and α=β=γ90°.
“Tetragonal Unit Cell” refers to a unit cell in which a≠b≠c; and α=β=γ90°.
“Trigonal/Rhombohedral Unit Cell” refers to a unit cell in which a≠b≠c; and α=β=γ≠90 °.
“Trigonal/Hexagonal Unit Cell” refers to a unit cell in which a=b=c; α=β=90°; and γ=120 °.
“Cubic Unit Cell” refers to a unit cell in which a=b=c; and α=β=γ90°.
“Crystal Lattice” refers to the array of points defined by the vertices of packed unit cells.
“Space Group” refers to the set of symmetry operations of a unit cell. In a space group designation (e.g., C2) the capital letter indicates the lattice type and the other symbols represent symmetry operations that can be carried out on the unit cell without changing its appearance.
“Asymmetric Unit” refers to the largest aggregate of molecules in the unit cell that possesses no symmetry elements that are part of the space group symmetry, but that can be juxtaposed on other identical entities by symmetry operations.
“Crystallogralhically-Related Dimer” refers to a dimer of two molecules wherein the symmetry axes or planes that relate the two molecules comprising the dimer coincide with the symmetry axes or planes of the crystal lattice.
“Non-Crystallogralphically-Related Dimer” refers to a dimer of two molecules wherein the symmetry axes or planes that relate the two molecules comprising the dimer do not coincide with the symmetry axes or planes of the crystal lattice.
“Isomorphous Replacement” refers to the method of using heavy-atom derivative crystals to obtain the phase information necessary to elucidate the three-dimensional structure of a crystallized polypeptide (Blundel et al., 1976, Protein Crystallography, Academic Press).
“Multi-Wavelength Anomalous Dispersion or MAD” refers to a crystallographic technique in which X-ray diffraction data are collected at several different wavelengths from a single heavy-atom derivative crystal, wherein the heavy atom has absorption edges near the energy of incoming X-ray radiation. The resonance between X-rays and electron orbitals leads to differences in X-ray scattering from absorption of the X-rays (known as anomalous scattering) and permits the locations of the heavy atoms to be identified, which in turn provides phase information for a crystal of a polypeptide. A detailed discussion of MAD analysis can be found in Hendrickson, 1985, Trans. Am. Crystallogr. Assoc. 21:11; Hendrickson et al., 1990, EMBO J. 9:1665; and Hendrickson, 1991, Science 4:91.
“Single Wavelength Anomalous Dispersion or SAD” refers to a crystallographic technique in which X-ray diffraction data are collected at a single wavelength from a single native or heavy-atom derivative crystal, and phase information is extracted using anomalous scattering information from atoms such as sulfur or chlorine in the native crystal or from the heavy atoms in the heavy-atom derivative crystal. The wavelength of X-rays used to collect data for this phasing technique need not be close to the absorption edge of the anomalous scatterer. A detailed discussion of SAD analysis can be found in Brodersen et al., 2000, Acta Cryst., D56:431-441.
“Single Isomorphous Replacement With Anomalous Scattering or SIRAS” refers to a crystallographic technique that combines isomorphous replacement and anomalous scattering techniques to provide phase information for a crystal of a polypeptide. X-ray diffraction data are collected at a single wavelength, usually from a single heavy-atom derivative crystal. Phase information obtained only from the location of the heavy atoms in a single heavy-atom derivative crystal leads to an ambiguity in the phase angle, which is resolved using anomalous scattering from the heavy atoms. Phase information is therefore extracted from both the location of the heavy atoms and from anomalous scattering of the heavy atoms. A detailed discussion of SIRAS analysis can be found in North, 1965, Acta Cryst. 18:212-216; Matthews, 1966, Acta Cryst. 20:82-86.
“Molecular Replacement” refers to the method of calculating initial phases for a new crystal of a polypeptide whose structure coordinates are unknown by orienting and positioning a polypeptide whose structure coordinates are known within the unit cell of the new crystal so as to best account for the observed diffraction pattern of the new crystal. Phases are then calculated from the oriented and positioned polypeptide and combined with observed amplitudes to provide an approximate Fourier synthesis of the structure of the polypeptides comprising the new crystal (Jones et al., 1991, Acta Crystallogr. 47:753-70; Brunger et al., 1998, Acta Crystallogr. D. Biol. Crystallogr. 54:905-21).
“Having substantially the same three-dimensional structure” refers to a polypeptide that is characterized by a set of atomic structure coordinates that have a root mean square deviation (r.m.s.d.) of less than or equal to about 2 Å when superimposed onto the atomic structure coordinates of Table 7 when at least about 50% to 100% of the Cα atoms of the coordinates are included in the superposition.
“Cα” As used herein, “Cα” refers to the alpha carbon of an amino acid residue.
A “neutralizing epitope” refers to an epitope of a flaviviral envelope protein that is specifically bound by an antibody that prevents effective infection by the flavivirus of a cell that would ordinarily be susceptible to infection by the flavivirus, when the antibody is present at neutralizing concentrations. One example of a neutralizing concentration of an antibody as determined by the plaque reduction neutralization test (PRNT50) is at least about 10 ng/ml. For example, a WNV neutralizing epitope is defined by the region, or a portion thereof, contacted by the monoclonal antibody E16.
An “enhancing epitope” refers to an epitope of a flaviviral envelope protein that is specifically bound by an antibody that enhances infection by a flavivirus of cells expressing an Fc receptor, e.g., the Fcy receptor, when the antibody is present at all concentrations in which antibody binding to virus occurs. It should be noted that neutralizing antibodies also can enhance infection, but only when their occupancy on the virion falls below a certain level.”
“Purified,” when used in relation to an antibody, refers to a composition of antibodies that each have substantially similar specificities; e.g., the antibodies in the composition each bind essentially the same epitope. One method to obtain a purified antibody is to affinity purify the antibody from a polyclonal antibody preparation using a molecule that comprises the epitope of interest but not undesirable epitope(s). For example, a molecule comprising a neutralizing epitope but not an enhancing epitope can be used to obtain a purified antibody that binds the neutralizing epitope that is substantially free (e.g., antibodies of other specificity constitute less than about 0.1% of the total preparation) of antibodies that specifically bind the enhancing epitope.
An “immune response” refers to one or more biological activities mediated by cells of the immune system in a subject. Such biological activities include, but are not limited to, production of antibodies; activation and proliferation of immune cells, such as, e.g., B cells, T cells, macrophages, leukocytes, lymphocytes, etc.; release of messenger molecules, such as cytokines, chemokines, interleukins, tumor necrosis factors, growth factors, etc.; and the like. An immune response is typically mounted when a cell of the immune system encounters non-self antigen that is recognized by a receptor present on the surface of the immune cell. The immune response preferably protects the subject to some degree against infection by a pathogen that bears the antigen against which the immune response is mounted.
An immune response may be “elicited,” “induced,” or “induced against” a particular antigen. Each of these terms is intended to be synonymous as used herein and refers to the ability of an immunogen to generate an immune response upon administration to a subject.
An “immunogen” is a molecule or combination of molecules that can induce an immune response in a subject when the immunogen is administered to the subject.
“Immunizing” refers to administering an immunogen to a subject.
An “effective amount” of an immunogen is an amount of the immunogen effective to elicit an immune response in a subject.
An “effective amount” of an antibody is an amount of the antibody that, when administered to a subject, is effective to treat, to prevent, or to ameliorate one or more symptoms of, infection by a pathogen expressing an epitope that the antibody specifically binds.
“Specific binding,” or other grammatical forms of this phrase, refers to the ability of a molecule to non-covalently interact with another molecule with some affinity. For example, an antibody, “specifically binds” a WNV E protein, when it binds that E protein with higher affinity than any other WNV protein. As another example, an antibody, “specifically binds” an epitope of a WNV E protein, when it binds that epitope with higher affinity than any other epitope of the WNV E protein.
A “linker” refers to a molecule that joins two other molecules, either covalently or non-covalently, e.g., through ionic, van der Waals or hydrogen bonds, such as, for example, a nucleic acid molecule that hybridizes to one complementary sequence at the 5′ end and to another complementary sequence at the 3′ end, thus joining two non-complementary sequences.
A “pharmaceutical composition” refers to a composition suitable for pharmaceutical use in a mammal. A pharmaceutical composition comprises a pharmacologically effective amount of an active agent and a pharmaceutically acceptable carrier. “Pharmacologically effective amount” refers to that amount of an agent effective to produce the intended pharmacological result. “Pharmaceutically acceptable carrier” refers to any of the standard pharmaceutical carriers, vehicles, diluents, buffers, and excipients, such as a phosphate buffered saline solution, 5% aqueous solution of dextrose, and emulsions, such as an oil/water or water/oil emulsion, and various types of wetting agents and/or adjuvants. Suitable pharmaceutical carriers and formulations are described in Remington's Pharmaceutical Sciences, 19th Ed. 1995, Mack Publishing Co., Easton. A “pharmaceutically acceptable salt” is a salt that can be formulated into a compound for pharmaceutical use including, e.g., metal salts (sodium, potassium, magnesium, calcium, etc.) and salts of ammonia or organic amines.
Preferred pharmaceutical carriers depend upon the intended mode of administration of the active agent. Typical modes of administration include enteral (e.g., oral, intranasal, rectal, or vaginal) or parenteral (e.g., subcutaneous, intramuscular, intravenous or intraperitoneal injection; or topical, transdermal, or transmucosal administration).
A “subject” of diagnosis, treatment, or administration is a human or non-human animal, including a mammal, such as a rodent (e.g., a mouse or rat), a lagomorph (e.g., a rabbit), or a primate. A subject of diagnosis, treatment, or administration is preferably a primate, and more preferably a human.
“Treatment” refers to therapeutic treatment. A “therapeutic” treatment is a treatment administered to a subject who exhibits signs of pathology for the purpose of diminishing, slowing the progression, eliminating, or halting those signs.
“Prevention” refers to prophylactic treatment. A “prophylactic” treatment is a treatment administered to a subject who does not exhibit signs of a disease or exhibits only early signs for the purpose of decreasing the risk of developing pathology.
“Hybridizing specifically to” or “specific hybridization” or “selectively hybridize to”, refers to the binding, duplexing, or hybridizing of a nucleic acid molecule preferentially to a particular nucleotide sequence under stringent conditions when that sequence is present in a complex mixture (e.g., total cellular) DNA or RNA.
The term “stringent conditions” refers to conditions under which a nucleic acid will hybridize preferentially to a nucleic acid having a complementary sequence, and to a lesser extent to, or not at all to, other sequences. “Stringent hybridization” and “stringent hybridization wash conditions” in the context of nucleic acid hybridization experiments such as Southern and northern hybridizations are sequence dependent, and are different under different environmental parameters. An extensive guide to the hybridization of nucleic acids can be found in Tijssen, 1993, Laboratory Techniques in Biochemistry and Molecular Biology—Hybridization with Nucleic Acid Probes, part I, chapter 2, “Overview of principles of hybridization and the strategy of nucleic acid probe assays”, Elsevier, N.Y.; Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory, 3rd ed., NY; and Ausubel et al., eds., Current Edition, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, N.Y.
Generally, highly stringent hybridization and wash conditions are selected to be about 5° C. lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength and pH) at which 50% of the target sequence hybridizes to a perfectly matched probe. Very stringent conditions are selected to be equal to the Tm for a particular probe.
One example of stringent hybridization conditions for hybridization of complementary nucleic acids which have more than about 100 complementary residues on a filter in a Southern or northern blot is 50% formalin with 1 mg of heparin at 42° C., with the hybridization being carried out overnight. An example of highly stringent wash conditions is 0.15 M NaCl at 72° C. for about 15 minutes. An example of stringent wash conditions is a 0.2× SSC wash at 65° C. for 15 minutes. See Sambrook et al. for a description of SSC buffer. A high stringency wash can be preceded by a low stringency wash to remove background probe signal. An exemplary medium stringency wash for a duplex of, e.g., more than about 100 nucleotides, is 1×SSC at 45° C. for 15 minutes. An exemplary low stringency wash for a duplex of, e.g., more than about 100 nucleotides, is 4-6×SSC at 40° C. for 15 minutes. In general, a signal to noise ratio of 2× (or higher) than that observed for an unrelated probe in the particular hybridization assay indicates detection of a specific hybridization.



4. BRIEF DESCRIPTION OF THE FIGURES
FIG. 1 provides the amino acid sequence of domain III of the WNV E protein identified with Genbank Accession No. AF196835, where amino acid 1 of FIG. 1 corresponds to amino acid 296 of WNV E protein (the amino acid sequence of domain III of WNV E protein is identified herein as SEQ ID NO:1; the amino acid sequence of WNV E protein is identified herein as SEQ ID NO:4);
FIG. 2 provides the amino acid sequence (SEQ ID NO:2)of the variable region of the light chain of monoclonal antibody E16, numbered according to the Kabat numbering system;
FIG. 3 provides the amino acid sequence (SEQ ID NO:3) of the variable region of the heavy chain of monoclonal antibody E16, numbered according to the Kabat numbering system;
FIG. 4 provides a diffraction pattern of the WNV DIII-E16 Fab complex, where the elution time of the WNV DIII-E16 Fab complex corresponds to a molecular weight of 60 kDa and the predicted molecular weight of the complex is 58 kDa;
FIG. 5 provides a Superdex 75 chromatogram of the WNV DIII-E16 Fab complex purification;
FIG. 6 provides a representative WNV DIII-E16 Fab crystal;
FIG. 7 provides an electron density map of the WNV DIII-E16 Fab complex contoured at 1.3σ;
FIG. 8 provides a Ramachandran plot of the WNV DIII-E16 Fab complex;
FIG. 9 provides a ribbon diagram of the structure of the WNV DIII-E16 Fab complex;
FIG. 10 provides a grasp diagram of the interface between the E16 Fab fragment and WNV DIII (the WNV DIII is rotated 180° with respect to the vertical line in the figure);
FIG. 11 provides a ribbon diagram of WNV DIII;
FIG. 12 provides flow cytometry diagrams showing binding of E16 and E21 to of yeast cells expressing wild-type or mutant versions of WNV DIII;
FIG. 13 provides diagrams showing detailed interactions of WNV DIII residues SerE306 and LysE307 with E16, with interfacial waters evident in the composite electron density omit map;
FIG. 14 provides a diagram showing interactions of ThrE330 and ThrE332 at the E16 interface;
FIG. 15 provides a set of alignments showing the sequence of the four segments of WNV DIII contacted by E16 aligned with the analogous residues of other flaviviruses, where the WNV DIII residues contacted by E16 are highlighted in light and dark gray, and deletions are indicated with a (#) (SEQ ID NOS: 5-44 are disclosed respectively in order of appearance);
FIG. 16 provides a ribbon diagram showing the structure of the WNV DIII dominant neutralizing epitope as defined by the E16/DIII complex;
FIG. 17 provides a diagram showing E16 docked onto the DEN-2 E dimer where (1OAN) through WNV DIII (“DIII”) super positioning indicates that binding likely occurs without WNV DI (“DI”) or WNV DII (gray ribbon below DI) contacts;
FIG. 18 provides a diagram showing E16 docked onto the post-fusion DEN-2 E trimer (1OK8), indicating accessibility of the binding site;
FIG. 19 provides a diagram showing the conserved structure of WNV DIII from the E16 complex (“DIII”), the pre-fusion DEN-2 (IOKE, light gray), and post-fusion DEN -2 (1OK8, dark gray), where the interaction of E16 with the flavivirus conserved TyrE302 in the N-terminal region of WNV DIII is highlighted;
FIG. 20 provides a diagram showing the E16 structural epitope mapped onto the cryo-electron microscopic reconstruction of the WNV virion presented as 2.0 Å radius Cα atoms;
FIG. 21A provides a diagram showing E16/WNV DIII complexes docked around the icosahedral three-fold axis, while FIG. 21B provides a diagram showing that WNV DIII situated around the outer ring of the five-fold axis is permissive to E16 binding, but the inner ring appears to exclude E16 engagement;
FIG. 22 presents a diagram showing that saturation binding by E16 is predicted to entail the binding of 120 of 180 WNV epitopes with exclusion of binding to the inner-five fold WNV DIIIs;
FIG. 23 presents graphs showing that two DI/DII specific neutralizing mAbs (E53 and E60) block cellular attachment significantly more than two DIII specific neutralizing antibodies (E16 and E24) or controls (no antibody, non-neutralizing mAb E22 or anti-SARS antibody ORF7a), reported as fold-reductions, with standard deviations, as the average of four to seven independent experiments performed in triplicate;
FIG. 24 presents graphs showing the dose-dependent blockade of WNV infection by E16 (black lines) and E53 (gray lines) in pre- and post-adsorption assays, reported as one of three representative experiments performed in duplicate;
FIG. 25 presents graphs showing that DIII specific mAbs effectively inhibit WNV infection of macrophages, while DI/DII specific E5 and E60 mAbs enhance infection. The data is one of three representative experiments performed in duplicate, with the dotted line representing the limit of sensitivity of the assay;
FIG. 26 presents graphs showing that pre-incubation with unlabeled mabs followed by addition of the APC-conjugates reveals that E16 and E60 both are self-competitive but not cross competitive for E binding; and
FIG. 27 presents a table summarizing the interactions between WNV DIII amino acids and E16 Fab amino acids.



4.1 BRIEF DESCRIPTION OF THE TABLES
Table 1 provides contact residues of WNV DIII and E16 Fab;
Table 2 provides hydrophobic contacts between WNV DIII and E16 Fab; and
Table 3 provides hydrogen bond contacts between WNV DIII and E16 Fab.
Table 4 provides representative flaviviruses and citations to the amino acid sequences of flaviviral envelope proteins;
Table 5 summarizes the X-ray crystallography data sets of WNV DIII-E16 Fab complex crystals that were used to determine the structures of the crystalline WNV DIII-E16 Fab complexes of the invention;
Table 6 summarizes the X-ray crystallography refinement parameters of the structures of crystalline WNV DIII-E16 Fab complexes of the invention; and
Table 7 provides the atomic structure coordinates of native WNV DIII-E16 Fab complex crystals of the invention as determined by X-ray crystallography. SEQ ID NOS: 45-53 are disclosed respectively in order of appearance.
5. DETAILED DESCRIPTION OF THE INVENTION
The present invention provides crystalline forms of polypeptide complexes corresponding to a Fab fragment of a neutralizing antibody in complex with domain III of the WNV E protein, an envelope protein obtained from a member of the viral family flaviviridae. Viruses from this family with related envelope proteins include, for example, WNV, Japanese encephalitis virus, Kunjin virus, Murray Valley encephalitis virus, Dengue virus, Montana myotis leukoencephalitis virus, Modoc virus, and Yellow Fever virus. In addition, the invention provides a three dimensional structure of such polypeptide complexes. Further, the invention provides methods of making high affinity antibodies that bind WNV DIII. Still further, the invention provides methods of identifying compounds that mimic WNV DIII and can be used to induce production of antibodies that recognize WNV DIII. In addition, the invention provides methods of identifying compounds that bind WNV DIII.
5.1 Crystalline WNV DIII-E16 Fab Complex
In one aspect, the invention provides crystals from which the atomic structure coordinates of the invention may be obtained, including native crystals and heavy-atom derivative crystals. Native crystals generally comprise substantially pure polypeptides corresponding to WNV DIII-E16 Fab complex in crystalline form.
It is to be understood that the crystalline WNV DIII-E16 Fab complex from which the atomic structure coordinates of the invention can be obtained is not limited to wild-type WNV DIII or wild-type E16 Fab. Indeed, the crystals may comprise mutants of wild-type WNV DIII or wild-type E16 Fab. Mutants of wild-type WNV DIII or wild-type E16 Fab are obtained by replacing at least one amino acid residue in the sequence of the wild-type WNV DIII or wild-type E16 Fab with a different amino acid residue, or by adding or deleting one or more amino acid residues within the wild-type sequence and/or at the N- and/or C-terminus of the wild-type WNV DIII or wild-type E16 Fab. Preferably, such mutants will crystallize under crystallization conditions that are substantially similar to those used to crystallize the wild-type WNV DIII-E16 Fab complex.
The types of mutants contemplated by this invention include conservative mutants, non-conservative mutants, deletion mutants, truncated mutants, extended mutants, methionine mutants, selenomethionine mutants, cysteine mutants and selenocysteine mutants. A mutant may have, but need not have, wild-type WNV DIII or wild-type E16 Fab binding activity. Preferably, a mutant displays biological activity that is substantially similar to that of the wild-type polypeptide. Methionine, selenomethionine, cysteine, and selenocysteine mutants are particularly useful for producing heavy-atom derivative crystals, as described in detail, below.
It will be recognized by one of skill in the art that the types of mutants contemplated herein are not mutually exclusive; that is, for example, a polypeptide having a conservative mutation in one amino acid may in addition have a truncation of residues at the N-terminus, and several Leu or Ile→Met mutations.
In addition, conservative or non-conservative amino acid substitutions can be made to amino acids of WNV DIII or E16 Fab that are implicated in association of WNV DIII with E16 Fab. Such conservative or non-conservative substitutions can affect, e.g., the affinity with which E16 Fab binds WNV DIII. In certain embodiments, the conservative or non-conservative amino acid substitutions can increase the affinity with which E16 Fab binds WNV DIII. In other embodiments, the conservative or non-conservative amino acid substitutions can decrease the affinity with which E16 Fab binds WNV DIII.
Conservative amino acid substitutions are well-known in the art, and include substitutions made on the basis of a similarity in polarity, charge, solubility, hydrophobicity and/or the hydrophilicity of the amino acid residues involved. Typical conservative substitutions are those in which the amino acid is substituted with a different amino acid that is a member of the same class or category, as those classes are defined herein. Thus, typical conservative substitutions include aromatic to aromatic, apolar to apolar, aliphatic to aliphatic, acidic to acidic, basic to basic, polar to polar, etc. Other conservative amino acid substitutions are well known in the art. It will be recognized by those of skill in the art that generally, a total of about 20% or fewer, typically about 10% or fewer, most usually about 5% or fewer, of the amino acids in the wild-type polypeptide sequence can be conservatively substituted with other amino acids without deleteriously affecting the biological activity and/or three-dimensional structure of the molecule, provided that such substitutions do not involve residues that are critical for activity, as discussed above.
In some embodiments, it may be desirable to make mutations in the active site of a protein, e.g., to reduce or completely eliminate protein activity. Mutations that will reduce or completely eliminate the activity of a particular protein will be apparent to those of skill in the art. For example, the amino acids identified in Tables 1 or 2 could be mutated in order to reduce or eliminate the binding activity of E16 Fab.
The amino acid residue Cys (C) is unusual in that it can form disulfide bridges with other Cys (C) residues or other sulfhydryl-containing amino acids (“cysteine-like amino acids”). The ability of Cys (C) residues and other cysteine-like amino acids to exist in a polypeptide in either the reduced free —SH or oxidized disulfide-bridged form affects whether Cys (C) residues contribute net hydrophobic or hydrophilic character to a polypeptide. While Cys (C) exhibits a hydrophobicity of 0.29 according to the consensus scale of Eisenberg (Eisenberg, 1984, supra), it is to be understood that for purposes of the present invention Cys (C) is categorized as a polar hydrophilic amino acid, notwithstanding the general classifications defined above. Preferably, Cys residues that are known to participate in disulfide bridges, such as those linking the heavy chain to the light chain of an antibody, or a portion thereof, are not substituted or are conservatively substituted with other cysteine-like amino acids so that the residue can participate in a disulfide bridge. Typical cysteine-like residues include, for example, Pen, hCys, etc. Substitutions for Cys residues that interfere with crystallization are discussed infra.
While in most instances the amino acids of WNV DIII and/or E16 Fab will be substituted with genetically-encoded amino acids, in certain circumstances mutants may include genetically non-encoded amino acids. For example, non-encoded derivatives of certain encoded amino acids, such as SeMet and/or SeCys, may be incorporated into the polypeptide chain using biological expression systems (such SeMet and SeCys mutants are described in more detail, infra).
Alternatively, in instances where the mutant will be prepared in whole or in part by chemical synthesis, virtually any non-encoded amino acids may be used, ranging from D-isomers of the genetically encoded amino acids to non-encoded naturally-occurring natural and synthetic amino acids.
Conservative amino acid substitutions for many of the commonly known non-genetically encoded amino acids are well known in the art. Conservative substitutions for other non-encoded amino acids can be determined based on their physical properties as compared to the properties of the genetically encoded amino acids.
In some instances, it may be particularly advantageous or convenient to substitute, delete from and/or add amino acid residues to WNV DIII and/or E16 Fab in order to provide convenient cloning sites in cDNA encoding the polypeptide, to aid in purification of the polypeptide, etc. Such substitutions, deletions and/or additions that do not substantially alter the three dimensional structure of the native WNV DIII and/or E16 Fab will be apparent to those having skills in the art. These substitutions, deletions and/or additions include, but are not limited to, His tags, BirA tags, intein-containing self-cleaving tags, maltose binding protein fusions, glutathione S-transferase protein fusions, antibody fusions, green fluorescent protein fusions, signal peptide fusions, biotin accepting peptide fulsions, and the like. In certain embodiments, the WNV DIII comprises a His tag. In other embodiments, the WNV DIII comprises a BirA tag. In a preferred embodiment, the WNV DIII comprises a His tag and a BirA tag.
Mutations may also be introduced into a polypeptide sequence where there are residues, e.g., cysteine residues, that interfere with crystallization. Such cysteine residues can be substituted with an appropriate amino acid that does not readily form covalent bonds with other amino acid residues under crystallization conditions; e.g., by substituting the cysteine with Ala, Ser or Gly. Any cysteine located in a non-helical or non-β-stranded segment, based on secondary structure assignments, are good candidates for replacement.
It should be noted that the mutants contemplated herein need not exhibit WNV DIII or E16 Fab binding activity. Indeed, amino acid substitutions, additions or deletions that interfere with the binding activity of WNV DIII and/or E16 Fab are specifically contemplated by the invention. Such crystalline polypeptides, or the atomic structure coordinates obtained therefrom, can be used to provide phase information to aid the determination of the three-dimensional X-ray structures of other related or non-related crystalline polypeptides.
The heavy-atom derivative crystals from which the atomic structure coordinates of the invention are obtained generally comprise a crystalline WNV DIII-E16 Fab complex in association with one or more heavy metal atoms. The polypeptide may correspond to a complex comprising wild-type WNV DIII and wild-type E16 Fab or a complex comprising mutant WNV DIII and/or E16 Fab, which may optionally be in complex with one or more molecules, as previously described. There are two types of heavy-atom derivatives of polypeptides: heavy-atom derivatives resulting from exposure of the protein to a heavy metal in solution, wherein crystals are grown in medium comprising the heavy metal, or in crystalline form, wherein the heavy metal diffuses into the crystal, and heavy-atom derivatives wherein the polypeptide comprises heavy-atom containing amino acids, e.g., selenomethionine and/or selenocysteine mutants.
In practice, heavy-atom derivatives of the first type can be formed by soaking a native crystal in a solution comprising heavy metal atom salts, or organometallic compounds, e.g., lead chloride, gold thiomalate, ethylmercurithiosalicylic acid-sodium salt (thimerosal), uranyl acetate, platinum tetrachloride, osmium tetraoxide, zinc sulfate, and cobalt hexamine, which can diffuise through the crystal and bind to the crystalline polypeptide complex.
Heavy-atom derivatives of this type can also be formed by adding to a crystallization solution comprising the polypeptide complex to be crystallized an amount of a heavy metal atom salt, which may associate with the protein complex and be incorporated into the crystal. The location(s) of the bound heavy metal atom(s) can be determined by X-ray diffraction analysis of the crystal. This information, in turn, is used to generate the phase information needed to construct the three-dimensional structure of the protein.
Heavy-atom derivative crystals may also be prepared from polypeptides that include one or more SeMet and/or SeCys residues (SeMet and/or SeCys mutants). Such selenocysteine or selenomethionine mutants may be made from complexes comprising wild-type WNV DIII and E16 Fab or complexes comprising mutant WNV DIII and/or E16 Fab by expression of WNV DIII or E16-encoding cDNAs in auxotrophic E. coli strains. Hendrickson et al., 1990, EMBO J. 9:1665-1672. In this method, the wild-type or mutant WNV DIII or cDNAs encoding the heavy and/or light chains of E16 may be expressed in a host organism on a growth medium depleted of either natural cysteine or methionine (or both) but enriched in selenocysteine or selenomethionine (or both). Alternatively, selenocysteine or selenomethionine mutants may be made using nonauxotrophic E. coli strains, e.g., by inhibiting methionine biosynthesis in these strains with high concentrations of Ile, Lys, Phe, Leu, Val or Thr and then providing selenomethionine in the medium (Doublie, 1997, Methods in Enzymology 276:523-530). Furthermore, selenocysteine can be selectively incorporated into polypeptides by exploiting the prokaryotic and eukaryotic mechanisms for selenocysteine incorporation into certain classes of proteins in vivo, as described in U.S. Pat. No. 5,700,660 to Leonard et al. (filed Jun. 7, 1995). One of skill in the art will recognize that selenocysteine is preferably not incorporated in place of cysteine residues that form disulfide bridges, as these may be important for maintaining the three-dimensional structure of the protein and are preferably not to be eliminated. One of skill in the art will further recognize that, in order to obtain accurate phase information, approximately one selenium atom should be incorporated for every 140 amino acid residues of the polypeptide chain. The number of selenium atoms incorporated into the polypeptide chain can be conveniently controlled by designing a Met or Cys mutant having an appropriate number of Met and/or Cys residues, as described more fully below.
In some instances, the polypeptide to be crystallized may not contain cysteine or methionine residues. For example, WNV DIII contains cysteine residues, but no methionine residues, while E16 Fab comprises methionine residues, but no cysteine residues that are not involved in intra- or intermolecular disulfide bonding. Therefore, if selenomethionine and/or selenocysteine mutants are to be used to obtain heavy-atom derivative crystals, methionine or cysteine residues may be introduced into the polypeptide chain of WNV DIII or E16 Fab, respectively.
Such mutations are preferably introduced into the polypeptide sequence at sites that will not disturb the overall protein fold. For example, a residue that is conserved among many members of the protein family or that is thought to be involved in maintaining its activity or structural integrity, as determined by, e.g., sequence alignments, should not be mutated to a Met or Cys. In addition, conservative mutations, such as Ser to Cys, or Leu or Ile to Met, are preferably introduced. One additional consideration is that, in order for a heavy-atom derivative crystal to provide phase information for structure determination, the location of the heavy atom(s) in the crystal unit cell should be determinable and provide phase information. Therefore, a mutation is preferably not introduced into a portion of the protein that is likely to be mobile, e.g., at, or within about 1-5 residues of, the N- and C-termini.
Conversely, if there are too many methionine and/or cysteine residues in a polypeptide sequence, over-incorporation of the selenium-containing side chains can lead to the inability of the polypeptide to fold and/or crystallize, and may potentially lead to complications in solving the crystal structure. In this case, methionine and/or cysteine mutants are prepared by substituting one or more of these Met and/or Cys residues with another residue. The considerations for these substitutions are the same as those discussed above for mutations that introduce methionine and/or cysteine residues into the polypeptide. Specifically, the Met and/or Cys residues are preferably conservatively substituted with Leu/Ile and Ser, respectively.
As DNA encoding cysteine and methionine mutants can be used in the methods described above for obtaining SeCys and SeMet heavy-atom derivative crystals, the preferred Cys or Met mutant will have one Cys or Met residue for every 140 amino acids.
5.2 Production of Polypeptides
The native and mutated WNV DIII and E16 polypeptides described herein may be chemically synthesized in whole or part using techniques that are well-known in the art (see, e.g., Creighton, 1983, Proteins: Structures and Molecular Principles, W.H. Freeman & Co., NY.). Alternatively, methods that are well known to those skilled in the art can be used to construct expression vectors containing the native or mutated WNV DIII and E16 polypeptide coding sequence and appropriate transcriptional/translational control signals. These methods include in vitro recombinant DNA techniques, synthetic techniques and in vivo recombination/genetic recombination. See, for example, the techniques described in the current editions of Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, 3d Ed., Cold Spring Harbor Laboratory, NY and Ausubel et al., 2004, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, NY.
A variety of host-expression vector systems may be utilized to express the WNV DIII and E16 coding sequences. These include but are not limited to microorganisms such as bacteria transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing the WNV DIII or E16 coding sequences; yeast transformed with recombinant yeast expression vectors containing the WNV DIII or E16 coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the WNV DIII or E16 coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus, CaMV; tobacco mosaic virus, TMV) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing the WNV DIII or E16 coding sequences; or animal cell systems. The expression elements of these systems vary in their strength and specificities.
Specifically designed vectors allow the shuttling of DNA between hosts such as bacteria-yeast or bacteria-animal cells. An appropriately constructed expression vector may contain: an origin of replication for autonomous replication in host cells, selectable markers, a limited number of useful restriction enzyme sites, a potential for high copy number, and active promoters. A promoter is defined as a DNA sequence that directs RNA polymerase to bind to DNA and initiate RNA synthesis. A strong promoter is one that causes mRNAs to be initiated at high frequency.
Depending on the host/vector system utilized, any of a number of suitable transcription and translation elements, including constitutive and inducible promoters, may be used in the expression vector. For example, when cloning in bacterial systems, inducible promoters such as the T7 promoter, pL of bacteriophage λ, plac, ptrp, ptac (ptrp-lac hybrid promoter) and the like may be used; when cloning in insect cell systems, promoters such as the baculovirus polyhedrin promoter may be used; when cloning in plant cell systems, promoters derived from the genome of plant cells (e.g., heat shock promoters; the promoter for the small subunit of RUBISCO; the promoter for the chlorophyll a/b binding protein) or from plant viruses (e.g., the 35S RNA promoter of CaMV; the coat protein promoter of TMV) may be used; when cloning in mammalian cell systems, promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter) may be used; when generating cell lines that contain multiple copies of the tyrosine kinase domain DNA, SV40-, BPV- and EBV-based vectors may be used with an appropriate selectable marker.
The expression vector may be introduced into host cells via any one of a number of techniques including but not limited to transformation, transfection, infection, protoplast fusion, and electroporation. The expression vector-containing cells are clonally propagated and individually analyzed to determine whether they produce WNV DIII or E16. Identification of WNV DIII- or E16-expressing host cell clones may be done by several means, including but not limited to immunological reactivity with anti-WNV DIII or anti-immunoglobulin antibodies, and the presence of host cell-associated WNV DIII or E16 biological activity.
Expression of WNV DIII or E16 cDNA may also be performed using in vitro produced synthetic mRNA. Synthetic mRNA can be efficiently translated in various cell-free systems, including but not limited to wheat germ extracts and reticulocyte extracts, as well as efficiently translated in cell based systems, including but not limited to microinjection into frog oocytes. Further, nucleic acids expressing WNV DIII or E16 cDNA can be constructed and expressed by gene synthesis using oligonucleotides. See Hoover & Lubkowski, 2002, Nucleic Acids Res 30:e43.
To determine the WNV DIII or E16 cDNA sequences that yields optimal levels of WNV DIII or E16 activity and/or WNV DIII or E16 protein, modified WNV DIII or E16 cDNA molecules are constructed. Host cells are transformed with the cDNA molecules and the levels of WNV DIII or E16 RNA and/or protein are measured.
Levels of WNV DIII or E16 protein in host cells are quantitated by a variety of methods such as immunoaffinity and/or ligand affinity techniques, WNV DIII- or E16-specific affinity beads or WNV DIII- or E16-specific antibodies are used to isolate 35s-methionine labeled or unlabeled WNV DIII or E16 protein. Labeled or unlabeled WNV DIII or E16 protein is analyzed by SDS-PAGE. Unlabeled WNV DIII or E16 is detected by Western blotting, ELISA or RIA employing WNV DIII- or E16-specific antibodies.
Following expression of WNV DIII or E16 in a recombinant host cell, WNV DIII or E16 may be recovered to provide WNV DIII or E16 in active form. Several WNV DIII or E16 purification procedures are available and suitable for use. Recombinant WNV DIII or E16 may be purified from cell lysates or from conditioned culture media, by various combinations of, or individual application of, fractionation, or chromatography steps that are known in the art.
In addition, recombinant WNV DIII can be separated from other cellular proteins by use of an immuno-affinity column made with monoclonal or polyclonal antibodies specific for full length nascent WNV DIII or polypeptide fragments thereof. The E16 monoclonal antibody is well-suited for use in such protocols; however, any antibody specific for WNV DIII can be used to immuno-purify WNV DIII. Similarly, recombinant WNV DIII can be used in an immuno-affinity column to purify E16. Further, other reagents that bind to antibodies, such as, e.g., protein A can be use to affinity-purify the E16 monoclonal antibody.
Alternatively, WNV DIII or E16 may be recovered from a host cell in an unfolded, inactive form, e.g., from inclusion bodies of bacteria. Proteins recovered in this form may be solublized using a denaturant, e.g., guanidinium hydrochloride, and then refolded into an active form using methods known to those skilled in the art, such as dialysis. See, for example, the techniques described in Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, 3d Ed., Cold Spring Harbor Laboratory, NY and Ausubel et al., 2004, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, NY.
Further, E16 can be prepared using a wide variety of techniques known in the art including the use of hybridoma, recombinant, and phage display technologies, or a combination thereof. For example, monoclonal antibodies can be produced using hybridoma techniques including those known in the art and taught, for example, in Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed. 1988); Hammerling, et al., in: Monoclonal Antibodies and T-Cell Hybridomas, pp. 563-681 (Elsevier, N.Y., 1981) (both of which are incorporated by reference in their entireties). The term “monoclonal antibody” as used herein is not limited to antibodies produced through hybridoma technology. The term “monoclonal antibody” refers to an antibody that is derived from a single clone, including any eukaryotic, prokaryotic, or phage clone, and not the method by which it is produced. Ascites fluid, which generally contains high levels of antibodies, can be generated by inoculating mice intraperitoneally with positive hybridoma clones.
Once a nucleic acid sequence encoding an antibody of the invention has been obtained according to standard techniques, the vector for the production of the antibody may be produced by recombinant DNA technology using techniques well known in the art. Methods which are well known to those skilled in the art can be used to construct expression vectors containing the antibody coding sequences and appropriate transcriptional and translational control signals. These methods include, for example, in vitro recombinant DNA techniques, synthetic techniques, and in vivo genetic recombination. (See, for example, the techniques described in Sambrook et al., 2001, Molecular Cloning, A Laboratory Manual, 3d Ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. and Ausubel et al. eds., 2004, Current Protocols in Molecular Biology, John Wiley & Sons, NY).
An expression vector comprising the nucleotide sequence of an antibody can be transferred to a host cell by conventional techniques (e.g., electroporation, liposomal transfection, and calcium phosphate precipitation) and the transfected cells are then cultured by conventional techniques to produce the antibody of the invention. In specific embodiments, the expression of the antibody is regulated by a constitutive, an inducible or a tissue, specific promoter.
The host cells used to express the recombinant antibodies of the invention may be either bacterial cells such as Escherichia coli, or, preferably, eukaryotic cells, especially for the expression of whole recombinant immunoglobulin molecule. In particular, mammalian cells such as Chinese hamster ovary cells (CHO), in conjunction with a vector such as the major intermediate early gene promoter element from human cytomegalovirus is an effective expression system for immunoglobulins (Foecking et al., 1998, Gene 45:101; Cockett et al., 1990, Bio/Technology 8:2).
A variety of host-expression vector systems may be utilized to express antibodies. Such host-expression systems represent vehicles by which the coding sequences of the antibodies may be produced and subsequently purified, but also represent cells which may, when transformed or transfected with the appropriate nucleotide coding sequences, express the antibodies of the invention in situ . These include, but are not limited to, microorganisms such as bacteria (e.g., E. coli and B. subtilis) transformed with recombinant bacteriophage DNA, plasmid DNA or cosmid DNA expression vectors containing immunoglobulin coding sequences; yeast (e.g., Saccharomyces Pichia) transformed with recombinant yeast expression vectors containing immunoglobulin coding sequences; insect cell systems infected with recombinant virus expression vectors (e.g., baculovirus) containing the immunoglobulin coding sequences; plant cell systems infected with recombinant virus expression vectors (e.g., cauliflower mosaic virus (CaMV) and tobacco mosaic virus (TMV)) or transformed with recombinant plasmid expression vectors (e.g., Ti plasmid) containing immunoglobulin coding sequences; or mammalian cell systems (e.g., COS, CHO, BHK, 293, 293T, 3T3 cells, lymphotic cells (see U.S. Pat. No. 5,807,715), Per C.6 cells (rat retinal cells developed by Crucell)) harboring recombinant expression constructs containing promoters derived from the genome of mammalian cells (e.g., metallothionein promoter) or from mammalian viruses (e.g., the adenovirus late promoter; the vaccinia virus 7.5K promoter).
In bacterial systems, a number of expression vectors may be advantageously selected depending upon the use intended for the antibody being expressed. For example, when a large quantity of such a protein is to be produced, for the generation of pharmaceutical compositions of an antibody, vectors which direct the expression of high levels of fusion protein products that are readily purified may be desirable. Such vectors include, but are not limited, to the E. coli expression vector pUR278 (Ruther et al., 1983, EMBO J. 2:1791), in which the antibody coding sequence may be ligated individually into the vector in frame with the lac Z coding region so that a fusion protein is produced; pIN vectors (Inouye & Inouye, 1985, Nucleic Acids Res. 13:3101-3109; Van Heeke & Schuster, 1989, J. Biol. Chem. 24:5503-5509); and the like. pGEX vectors may also be used to express foreign polypeptides as fusion proteins with glutathione S-transferase (GST). In general, such fusion proteins are soluble and can easily be purified from lysed cells by adsorption and binding to a matrix glutathione-agarose beads followed by elution in the presence of free glutathione. The pGEX vectors are designed to include thrombin or factor Xa protease cleavage sites so that the cloned target gene product can be released from the GST moiety.
In an insect system, Autographa californica nuclear polyhedrosis virus (AcNPV) is used as a vector to express foreign genes. The virus grows in Spodoptera frugiperda cells. The antibody coding sequence may be cloned individually into non-essential regions (e.g., the polyhedrin gene) of the virus and placed under control of an AcNPV promoter (e.g., the polyhedrin promoter).
In mammalian host cells, a number of viral-based expression systems may be utilized. In cases where an adenovirus is used as an expression vector, the antibody coding sequence of interest may be ligated to an adenovirus transcription/translation control complex, e.g., the late promoter and tripartite leader sequence. This chimeric gene may then be inserted in the adenovirus genome by in vitro or in vivo recombination. Insertion in a non-essential region of the viral genome (e.g., region E1 or E3) will result in a recombinant virus that is viable and capable of expressing the immunoglobulin molecule in infected hosts (e.g., see Logan & Shenk, 1984, Proc. Natl. Acad. Sci. USA 81:355-359). Specific initiation signals may also be required for efficient translation of inserted antibody coding sequences. These signals include the ATG initiation codon and adjacent sequences. Furthermore, the initiation codon must be in phase with the reading frame of the desired coding sequence to ensure translation of the entire insert. These exogenous translational control signals and initiation codons can be of a variety of origins, both natural and synthetic. The efficiency of expression may be enhanced by the inclusion of appropriate transcription enhancer elements, transcription terminators, etc. (see Bittner et al., 1987, Methods in Enzymol. 153:51-544).
In addition, a host cell strain may be chosen which modulates the expression of the inserted sequences, or modifies and processes the gene product in the specific fashion desired. Such modifications (e.g., glycosylation) and processing (e.g., cleavage) of protein products may be important for the finction of the protein. Different host cells have characteristic and specific mechanisms for the post-translational processing and modification of proteins and gene products. Appropriate cell lines or host systems can be chosen to ensure the correct modification and processing of the foreign protein expressed. To this end, eukaryotic host cells which possess the cellular machinery for proper processing of the primary transcript, glycosylation, and phosphorylation of the gene product may be used. Such mammalian host cells include but are not limited to CHO, VERY, BHK, Hela, COS, MDCK, 293, 293T, 3T3, WI38, BT483, Hs578T, HTB2, BT20 and T47D, CRL7030 and Hs578Bst.
For long-term, high-yield production of recombinant proteins, stable expression is preferred. For example, cell lines which stably express an antibody of the invention may be engineered. Rather than using expression vectors which contain viral origins of replication, host cells can be transformed with DNA controlled by appropriate expression control elements (e.g., promoter, enhancer, sequences, transcription terminators, polyadenylation sites, etc.), and a selectable marker. Following the introduction of the foreign DNA, engineered cells may be allowed to grow for 1-2 days in an enriched media, and then are switched to a selective media. The selectable marker in the recombinant plasmid confers resistance to the selection and allows cells to stably integrate the plasmid into their chromosomes and grow to form foci which in turn can be cloned and expanded into cell lines. This method may advantageously be used to engineer cell lines which express the antibodies of the invention. Such engineered cell lines may be particularly useful in screening and evaluation of compounds that interact directly or indirectly with the antibodies of the invention.
A number of selection systems may be used, including but not limited to the herpes simplex virus thymidine kinase (Wigler et al., 1977, Cell 11:223), hypoxanthine-guanine phosphoribosyltransferase (Szybalska & Szybalski, 1992, Proc. Natl. Acad. Sci. USA 48:202), and adenine phosphoribosyltransferase (Lowy et al., 1980, Cell 22:817) genes can be employed in tk-, hgprt- or aprt- cells, respectively. Also, antimetabolite resistance can be used as the basis of selection for the following genes: dhfr, which confers resistance to methotrexate (Wigler et al., 1980, Proc. Natl. Acad. Sci. USA 77:357; O'Hare et al., 1981, Proc. Natl. Acad. Sci. USA 78:1527); gpt, which confers resistance to mycophenolic acid (Mulligan & Berg, 1981, Proc. Natl. Acad. Sci. USA 78:2072); neo, which confers resistance to the aminoglycoside G-418 Clinical Pharmacy 12:488-505; Wu and Wu, 1991, 3:87-95; Tolstoshev, 1993, Ann. Rev. Pharmacol. Toxicol. 32:573-596; Mulligan, 1993, Science 260:926-932; Morgan and Anderson, 1993, Ann. Rev. Biochem. 62:191-217; and May, 1993, TIB TECH 11:155-215); and hygro, which confers resistance to hygromycin (Santerre et al., 1984, Gene 30:147). Methods commonly known in the art of recombinant DNA technology which can be used are described in Ausubel et al. (eds.), 2004, Current Protocols in Molecular Biology, John Wiley & Sons, NY; Kriegler, 1990, Gene Transfer and Expression, A Laboratory Manual, Stockton Press, NY; and in Chapters 12 and 13, Dracopoli et al. (eds), 1994, Current Protocols in Human Genetics, John Wiley & Sons, NY.; and Colberre-Garapin et al., 1981, J Mol. Biol. 150:1.
The expression levels of an antibody can be increased by vector amplification (for a review, see Bebbington and Hentschel, The use of vectors based on gene amplification for the expression of cloned genes in mammalian cells in DNA cloning, Vol.3. (Academic Press, New York, 1987)). When a marker in the vector system expressing an antibody is amplifiable, increase in the level of inhibitor present in culture of host cell can increase the number of copies of the marker gene. Since the amplified region is associated with the nucleotide sequence of the antibody, production of the antibody can also increase. See Crouse et al., 1983, Mol. Cell. Biol. 3:257.
The host cell may be co-transfected with two expression vectors of the invention, the first vector encoding a heavy chain derived polypeptide and the second vector encoding a light chain derived polypeptide. The two vectors may contain identical selectable markers which enable equal expression of heavy and light chain polypeptides. Alternatively, a single vector may be used which encodes both heavy and light chain polypeptides. In such situations, the light chain should be placed before the heavy chain to avoid an excess of toxic free heavy chain. See Proudfoot, 1986, Nature 322:52; and Kohler, 1980, Proc. Natl. Acad. Sci. USA 77:2197. The coding sequences for the heavy and light chains may comprise cDNA or genomic DNA.
Once the antibody of the invention has been recombinantly expressed, it may be purified by any method known in the art for purification of an antibody, for example, by chromatography (e.g., ion exchange, affinity, particularly by affinity for the specific antigen after Protein A, and sizing column chromatography), centrifugation, differential solubility, or by any other standard technique for the purification of proteins.
Still further, Fab fragments of E16 can be prepared from E16 according to any known method without limitation. Generally, Fab fragments of E16 are prepared by Papain digestion of E16; however, any technique that cleaves the E16 heavy chain at or near the hinge region can be used to prepare the Fab fragments. Repetitive protocols for making Fab fragments from antibodies, including monoclonal antibodies, are described in, e.g., Harlow et al., 1988, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory Press, 2nd ed. These techniques can be used to prepare Fab fragments from E16 prepared according to any of the methods described herein.
5.3 Crystallization of Polypeptides and Characterization of Crystal
The native, heavy-atom derivative, and/or poly-crystals from which the atomic structure coordinates of the invention can be obtained by conventional means as are well-known in the art of protein crystallography, including batch, liquid bridge, dialysis, and vapor diffusion methods (see, e.g., McPherson, 1998, Crystallization of Biological Macromolecules, Cold Spring Harbor Press, New York; McPherson, 1990, Eur. J. Biochem. 189:1-23.; Weber, 1991, Adv. Protein Chem. 41:1-36).
Generally, native crystals are grown by dissolving substantially pure WNV DIII-E16 Fab complex in an aqueous buffer containing a precipitant at a concentration just below that necessary to precipitate the protein. Examples of precipitants include, but are not limited to, polyethylene glycol, ammonium sulfate, 2-methyl-2,4-pentanediol, sodium citrate, sodium chloride, glycerol, isopropanol, lithium sulfate, sodium acetate, sodium formate, potassium sodium tartrate, ethanol, hexanediol, ethylene glycol, dioxane, t-butanol and combinations thereof. Water is removed by controlled evaporation to produce precipitating conditions, which are maintained until crystal growth ceases.
In a preferred embodiment, native crystals are grown by vapor diffuision in sitting drops (McPherson, 1982, Preparation and Analysis of Protein Crystals, John Wiley, New York; McPherson, 1990, Eur. J. Biochem. 189:1-23). In this method, the polypeptide/precipitant solution is allowed to equilibrate in a closed container with a larger aqueous reservoir having a precipitant concentration optimal for producing crystals. Generally, less than about 25 μL of substantially pure polypeptide solution is mixed with an equal volume of reservoir solution, giving a precipitant concentration about half that required for crystallization. The sealed container is allowed to stand, usually for about 2-6 weeks, until crystals grow.
For native crystals from which the atomic structure coordinates of the invention are obtained, it has been found that hanging drops of about 1.5 μL containing about 0.5 μL of 10-15 mg/ml WNV DIII-E16 Fab complex in 20 mM HEPES (pH 7.5) and 0.01% NaN3, about 0.5 μl of 0.1 M HEPES (pH 8.5), 19% PEG 4000, 0.1M glycine and 0.5 μl of deionized water over wells containing precipitant solution containing 100 mM HEPES (pH 8.5), 19% PEG 4000, and 100 mM glycine for about one week at 20° C. provide diffraction quality crystals.
In some embodiments in accordance with the present invention, a native crystal may be grown using a microbatch method. The microbatch crystallization method was originally developed to carry out protein crystallization by Douglas Instruments Ltd (Berkshire, United Kingdom) in collaboration with Imperial College, London. The method was developed to allow theoretical studies but can be used for routine large scale crystallization, since it is very rapid and uses only about as little as 0.1 to 1 μl of protein per trial. Like the original batch crystallization methods that were used in the early days of protein crystallization, the microbatch method involves the simple combination of protein with precipitants, buffers, etc., generally without any subsequent concentration step. The ingredients are simply mixed at their final concentrations. Because very small volumes are used, the droplets are generally covered, e.g., with paraffin oil, to prevent evaporation. Vapor Plates designed for batch crystallization available from Douglas Instruments can be used in such methods. These have 96 wells, each holding about 9 μl. Droplets with volumes from about 0.2 to about 2 μl are dispensed at the bottom of the wells. With a special microtip and highly accurate motorized syringes, very small droplets can be dispensed accurately. The dispensing error is generally around 20 nl.
Microbatch can be more effective than vapor diffusion for screening: for a given amount of time and material, more hits are usually found using the microbatch crystallization method (Baldock et al., 1996, Journal of Crystal Growth 168:170-174). Microbatch gives superior crystals for data collection in about 50% of proteins. In one example, a protein unstable to temperature changes, which couldn't be crystallized under vapor diffusion conditions, was crystallized using a microbatch method (Conti et al., 1996, Acta Cryst. D52:876-878). Microbatch can be used for controlling nucleation by carefully varying the temperature (Blow et al., 1994, Protein Science 3:1638-43). A final advantage is that the thick skins that often form with vapor diffusion are eliminated (e.g. Pearl et al., 1994, EMBO J. 13:5810-5817). Microbatch complements, rather than replaces, vapor diffusion. The main disadvantage of microbatch is that it can be relatively difficult to change conditions using the same protein sample, but it is particularly useful for screening of crystallization conditions.
Of course, those having skill in the art will recognize that the above-described crystallization conditions can be varied. Such variations may be used alone or in combination, and include polypeptide solutions containing polypeptide concentrations between 0.01 mg/mL and 100 mg/mL, HEPES concentrations between 0.1 mM and 500 mM, glycine concentrations between 0.001 mM and 100 mM, pH ranges between 4.0 and 12.0; and reservoir solutions containing PEG 4000 concentrations between about 0.1% and 50% (w/v), polyethylene glycol of other molecular weights between 0.01% and 50.0% (v/v), NaN3 concentrations between 0.001% and 10% (w/v), and temperature ranges between 5° C. and 40° C. Other buffer solutions may be used such as BIS-TRIS, MES, MOPS, MOPSO, PIPES, TRIS, and the like, so long as the desired pH range is maintained.
In some embodiments in accordance with the present invention, crystals are obtained by methods of high-throughput crystallization (HTC).
Heavy-atom derivative crystals can be obtained by soaking native crystals in mother liquor containing salts of heavy metal atoms.
Heavy-atom derivative crystals can also be obtained from SeMet and/or SeCys mutants, as described above for native crystals.
Mutant proteins may crystallize under slightly different crystallization conditions than wild-type protein, or under very different crystallization conditions, depending on the nature of the mutation, and its location in the protein. For example, a non-conservative mutation may result in alteration of the hydrophilicity of the mutant, which may in turn make the mutant protein either more soluble or less soluble than the wild-type protein. Typically, if a protein becomes more hydrophilic as a result of a mutation, it will be more soluble than the wild-type protein in an aqueous solution and a higher precipitant concentration will be needed to cause it to crystallize. Conversely, if a protein becomes less hydrophilic as a result of a mutation, it will be less soluble in an aqueous solution and a lower precipitant concentration will be needed to cause it to crystallize. If the mutation happens to be in a region of the protein involved in crystal lattice contacts, crystallization conditions may be affected in more unpredictable ways.
WNV DIII-E16 Fab complex crystals can be obtained by soaking a WNV DIII crystal in mother liquor containing E16 Fab, by soaking an E16 Fab crystal in mother liquor containing WNV DIII, or by co-crystallizing WNV DIII-E16 Fab complex.
5.4 Characterization of Crystals
The dimensions of a unit cell of a crystal are defined by six numbers, the lengths of three unique edges, a, b, and c, and three unique angles, α, β, and γ. The type of unit cell that comprises a crystal is dependent on the values of these variables, as discussed above in Section 3.2. While the following discussion relates to solving the structure of the compounds constituting the crystal by X-ray diffraction, one skilled in the art will recognize that other methods (e.g. Laue, electron or neutron diffraction) could also be used. Such methods are also intended to be within the scope of the present invention.
When a crystal is placed in an X-ray beam, the incident X-rays interact with the electron cloud of the molecules that make up the crystal, resulting in X-ray scatter. The combination of X-ray scatter with the lattice of the crystal gives rise to nonuniformity of the scatter; areas of high intensity are called diffracted X-rays. The angle at which diffracted beams emerge from the crystal can be computed by treating diffraction as if it were reflection from sets of equivalent, parallel planes of atoms in a crystal (Bragg's Law). The most obvious sets of planes in a crystal lattice are those that are parallel to the faces of the unit cell. These and other sets of planes can be drawn through the lattice points. Each set of planes is identified by three indices, hkl. The h index gives the number of parts into which the a edge of the unit cell is cut, the k index gives the number of parts into which the b edge of the unit cell is cut, and the l index gives the number of parts into which the c edge of the unit cell is cut by the set of hkl planes. Thus, for example, the 235 planes cut the a edge of each unit cell into halves, the b edge of each unit cell into thirds, and the c edge of each unit cell into fifths. Planes that are parallel to the bc face of the unit cell are the 100 planes; planes that are parallel to the ac face of the unit cell are the 010 planes; and planes that are parallel to the ab face of the unit cell are the 001 planes.
When a detector is placed in the path of the diffracted X-rays, in effect cutting into the sphere of diffraction, a series of spots, or reflections, are recorded to produce a “still” diffraction pattern. Each reflection is the result of X-rays reflecting off one set of parallel planes, and is characterized by an intensity, which is related to the distribution of molecules in the unit cell, and hkl indices, which correspond to the parallel planes from which the beam producing that spot was reflected. If the crystal is rotated about an axis perpendicular to the X-ray beam, a large number of reflections is recorded on the detector, resulting in a diffraction pattern as shown, for example, in FIG. 4.
The unit cell dimensions and space group of a crystal can be determined from its diffraction pattern. First, the spacing of reflections is inversely proportional to the lengths of the edges of the unit cell. Therefore, if a diffraction pattern is recorded when the X-ray beam is perpendicular to a face of the unit cell, two of the unit cell dimensions may be deduced from the spacing of the reflections in the x and y directions of the detector, the crystal-to-detector distance, and the wavelength of the X-rays. Those of skill in the art will appreciate that, in order to obtain all three unit cell dimensions, the crystal can be rotated such that the X-ray beam is perpendicular to another face of the unit cell. Second, the angles of a unit cell can be determined by the angles between lines of spots on the diffraction pattern. Third, the absence of certain reflections and the repetitive nature of the diffraction pattern, which may be evident by visual inspection, indicate the internal symmetry, or space group, of the crystal. Therefore, a crystal may be characterized by its unit cell and space group, as well as by its diffraction pattern.
Once the dimensions of the unit cell are determined, the likely number of polypeptides in the asymmetric unit can be deduced from the size of the polypeptide, the density of the average protein, and the typical solvent content of a protein crystal, which is usually in the range of 30-70% of the unit cell volume (Matthews, 1968, J. Mol. Biol. 33:491-497).
The WNV DIII-E16 Fab complex crystals of the present invention are generally characterized by a diffraction pattern, as shown in FIG. 4. The crystals are further characterized by unit cell dimensions and space group symmetry information obtained from the diffraction patterns, as described above. The crystals, which may be native crystals, heavy-atom derivative crystals or poly-crystals, have a orthorhombic unit cell (i.e., unit cells wherein a≠b≠c and α=β=γ90°) and space group symmetry P212121.
One form of crystalline WNV DIII-E16 Fab complex was obtained. In this form (designated “P212121 form”), the unit cell has dimensions of a=52.4+/−0.2 Å, b=83.3 +/−0.2 Å, c=110.6+/ 0.2 Å. There are believed to be 3 molecules in the asymmetric unit: one molecule of WNV DIII, one light chain of E16, and one papain-cleaved heavy chain of E16.
5.5 Collection of Data and Determination of Structure Solutions
The diffraction pattern of a crystal is related to the three-dimensional shape of the molecules that constitute the crystal by a Fourier transform. It has been established that diffraction patterns of a crystal can result from X-ray diffraction as well as Laue, electron or neutron diffraction. X-ray diffraction has been the most widely used methods for determining macromolecular structures. It is therefore used in this section by way of illustration to discuss the processes of diffraction data collection and subsequent structure determination. The scope of the present invention is, however, by no means limited only to X-ray diffraction analyses of protein crystals. After enough diffraction data are collected for a crystal, the process of determining the solution is in essence a re-focusing of the diffracted X-rays to produce a three-dimensional image of the molecule in the crystal. Since lenses capable of focusing X-ray radiation do not yet exist, the structure determination can be done via mathematical operations that simulate the re-focusing process.
The sphere of diffraction has symmetry that depends on the internal symmetry of the crystal, which means that certain orientations of the crystal will produce the same set of reflections. Thus, a crystal with high symmetry has a more repetitive diffraction pattern, and there are fewer unique reflections that need to be recorded in order to have a complete representation of the diffraction. The goal of data collection, a dataset, is a set of consistently measured, indexed intensities for as many reflections as possible. A complete dataset is collected if at least 80%, preferably at least 90%, most preferably at least 95% of unique reflections are recorded. In one embodiment, a complete dataset is collected using one crystal. In another embodiment, a complete dataset is collected using more than one crystal of the same type.
Sources of X-rays include, but are not limited to, a rotating anode X-ray generator such as a Rigaku RU-200 or a beamline at a synchrotron light source, such as the Advanced Photon Source at Argonne National Laboratory, the Advanced Light Source at the Lawrence Berkeley Laboratory, and the Stanford Synchrotron Radiation Laboratory at the Stanford Linear Acceleration Center. Suitable detectors for recording diffraction patterns include, but are not limited to, X-ray sensitive film, multiwire area detectors, image plates coated with phosphorus, and CCD cameras. Typically, the detector and the X-ray beam remain stationary, so that, in order to record diffraction from different parts of the crystal's sphere of diffraction, the crystal itself is moved via an automated system of moveable circles called a goniostat.
One of the biggest problems in data collection, particularly from macromolecular crystals having a high solvent content, is the rapid degradation of the crystal in the X-ray beam. In order to slow the degradation, data is often collected from a crystal at liquid nitrogen temperatures. In order for a crystal to survive the initial exposure to liquid nitrogen, the formation of ice within the crystal can be prevented by the use of a cryoprotectant. Suitable cryoprotectants include, but are not limited to, low molecular weight polyethylene glycols, ethylene glycol, sucrose, glycerol, xylitol, and combinations thereof. Crystals may be soaked in a solution comprising the one or more cryoprotectants prior to exposure to liquid nitrogen, or the one or more cryoprotectants may be added to the crystallization solution. Data collection at liquid nitrogen temperatures may allow the collection of an entire dataset from one crystal.
Once a dataset is collected, the information is used to determine the three-dimensional structure of the molecule in the crystal. However, this cannot be done from a single measurement of reflection intensities because certain information, known as phase information, is lost between the three-dimensional shape of the molecule and its Fourier transform, the diffraction pattern. This phase information can be acquired by methods described below in order to perform a Fourier transform on the diffraction pattern to obtain the three-dimensional structure of the molecule in the crystal. It is the determination of phase information that in effect refocuses X-rays to produce the image of the molecule.
One method of obtaining phase information is by isomorphous replacement, in which heavy-atom derivative crystals are used. In this method, diffraction data for both heavy-atom derivative crystals and native crystals are collected. Differences in diffraction patterns between the native and derivative datasets can be used to determine the positions of heavy atoms bound to the molecules in the heavy-atom derivative crystal. This information can then be used to obtain the phase information necessary to elucidate the three-dimensional structure of the material that constitutes the native crystals (Blundel et al., 1976, Protein Crystallography, Academic Press). In more recent applications of the isomorphous replacement method, manual and automatic (as implemented by the program SHELX) search procedures have been applied to locate the position of the heavy atoms in the derivative crystals (Sheldrick et al., 1993, Acta Cryst. D49:18-23). In other recent applications of the isomorphous replacement method, the inert gas Xenon is introduced into a native crystal to form a heavy atom derivative crystal. Xenon atoms occupy holes in a protein molecule through pure Van der Waals interaction. Examples of isomorphous replacement by Xenon derivatized crystals can be found in Sauer et al., 1997, J. Appl. Cryst. 30:476-486 and Panjikar and Tucker, 2002, Acta Cryst. D58:1413-1420.
Another method of obtaining phase information is by molecular replacement, which is a method of calculating initial phases for a new crystal of a polypeptide whose structure coordinates are unknown by orienting and positioning a polypeptide whose structure coordinates are known within the unit cell of the new crystal so as to best account for the observed diffraction pattern of the new crystal. Phases are then calculated from the oriented and positioned polypeptide and combined with observed amplitudes to provide an approximate Fourier synthesis of the structure of the molecules comprising the new crystal. (Lattman, 1985, Methods in Enzymology 115:55-77; Rossmann, 1972, “The Molecular Replacement Method,” Int. Sci. Rev. Ser. No. 13, Gordon & Breach, New York). The molecular replacement method can be used when a protein with unknown structure shares a certain degree of sequence homology with a protein whose structure is already known. Conventional molecular replacement methods comprise two search algorithms: a rotational search finction and a translational search function. Molecular replacement methods can be found in many existing computer programs such as AMoRe (Navaza, 1994, Acta, Cryst. A50:157-163) CNS (Brunger et al., 1998, Acta Cryst. D54:905-921), as well as many programs in the CCP4 package suites (Collaborative Computational Project, Number 4, 1994).
A third method of phase determination is multi-wavelength anomalous diffraction or MAD. In this method, X-ray diffraction data are collected at several different wavelengths from a single crystal containing at least one heavy atom with absorption edges near the energy of incoming X-ray radiation. The resonance between X-rays and electron orbitals leads to differences in X-ray scattering that permits the locations of the heavy atoms to be identified, which in turn provides phase information for a crystal of a polypeptide. MAD analysis uses a radiation source with capacity to adjust its output wavelength. Nearly all synchrotron source around the world are now equipped with the capacity. A detailed discussion of MAD analysis can be found in Hendrickson, 1985, Trans. Am. Crystallogr. Assoc. 21:11; Hendrickson et al., 1990, EMBO J. 9:1665-1672; and Hendrickson, 1991, Science 4:91. In the traditional approach, Se atoms (atomic number 34, in the same group at sulfur), usually in the form of Se-Met, are introduced into native protein prior to crystallization to add anomalous scattering property to the protein crystal (Hendrickson et al, 1990, EMBO J. 9:1665-1672; Leahy et al,. 1992, Science, 258:987-991). Incorporating Se-Met into protein is usually achieved by growing recombinant vectors in the presence of medium containing Se-Met supplement (Dyer et al., 2005, Protein Sci. 14:1508-1517).
A fourth method of determining phase information is single wavelength anomalous dispersion or SAD. In this technique, X-ray diffraction data are collected at a single wavelength from a single native or heavy-atom derivative crystal, and phase information is extracted using anomalous scattering information from atoms such as sulfur or chlorine in the native crystal or from the heavy atoms in the heavy-atom derivative crystal. The wavelength of X-rays used to collect data for this phasing technique need not be close to the absorption edge of the anomalous scatterer. A detailed discussion of SAD analysis can be found in Brodersen et al., 2000, Acta Cryst. D56:431-441. SAD eliminates the requirement for a radiation source with adjustable wavelengths. It is possible to utilize non-synchrotron radiation to determine protein structures by anomalous scattering. For example, the structure of human formylglycine-generating enzyme was determined by de novo calcium and sulfur SAD phasing at a non-synchrotron radiation source (Roeser et al., 2005, Acta Cryst. D61:1057-1066).
A fifth method of determining phase information is single isomorphous replacement with anomalous scattering or SIRAS. This technique combines isomorphous replacement and anomalous scattering techniques to provide phase information for a crystal of a polypeptide. X-ray diffraction data are collected at a single wavelength, usually from a single heavy-atom derivative crystal. Phase information obtained only from the location of the heavy atoms in a single heavy-atom derivative crystal leads to an ambiguity in the phase angle, which is resolved using anomalous scattering from the heavy atoms. Phase information is therefore extracted from both the location of the heavy atoms and from anomalous scattering of the heavy atoms. A detailed discussion of SIRAS analysis can be found in North, 1965, Acta Cryst. 18:212-216; Matthews, 1966, Acta Cryst. 20:82-86. It is possible to combine the techniques of MAD and SAD phasing with SIRAS and determine protein structure without synchrotron radiation. For example, the structure of E. coli argininosuccinate synthetase was determined using Cu-Kappa radiation in a non-synchrotron source with S-SAD, Se-SAD and S/Se-SIRAS phasing techniques (Lenike et al., 2002, Acta Cryst. D58:2096-2101).
Methods for phase determination have been discussed individually for the purpose of clear illustration. It is necessary to emphasize again that these methods are often combined in practice as previously stated. For example, the methods of MAD, SAD, and SIRAS were all explored when the structure of human mannose-6-phosphate/insulin-like growth factor II receptor was determined (Uson et al., 2002, Acta Cryst. D59:57-66). Also in this study, halide atoms, e.g. bromide and iodide as well as sulfur were used in extract the overall phase information of the molecule instead of the standard Se-Met MAD or SAD phasing techniques.
Once phase information is obtained, it is combined with the diffraction data to produce an electron density map, an image of the electron clouds that surround the molecules in the unit cell. The higher the resolution of the data, the more distinguishable are the features of the electron density map, e.g., amino acid side chains and the positions of carbonyl oxygen atoms in the peptide backbones, because atoms that are closer together are resolvable. A model of the macromolecule is then built into the electron density map with the aid of a computer, using as a guide all available information, such as the polypeptide sequence and the established rules of molecular structure and stereochemistry. Interpreting the electron density map is a process of finding the chemically reasonable conformation that fits the map precisely.
After a model is generated, a structure is refined. Refinement is the process of minimizing the function Φ, which is the difference between observed and calculated intensity values (measured by an R-factor), and which is a function of the position, temperature factor, and occupancy of each non-hydrogen atom in the model. This usually involves alternate cycles of real space refinement, i.e., calculation of electron density maps and model building, and reciprocal space refinement, i.e., computational attempts to improve the agreement between the original intensity data and intensity data generated from each successive model. Refinement ends when the function Φ converges on a minimum wherein the model fits the electron density map and is stereochemically and conformationally reasonable. During refinement, ordered solvent molecules are added to the structure.
5.5.1 Structures of WNV DIII-E16 Fab Complex
The present invention provides, for the first time, the high-resolution three-dimensional structures and atomic structure coordinates of crystalline WNV DIII-E16 Fab complex as determined by X-ray crystallography. The specific methods used to obtain the structure coordinates are provided in the examples, infra. The atomic structure coordinates of crystalline WNV DIII-E16 Fab complex, obtained from the P212121 form of the crystal to 2.5 Å resolution, are listed in Table 7.
Those having skill in the art will recognize that atomic structure coordinates as determined by X-ray crystallography are not without error. Thus, it is to be understood that any set of structure coordinates obtained for crystals of WNV DIII-E16 Fab complex, whether native crystals, heavy-atom derivative crystals or poly-crystals, that have a root mean square deviation (“r.m.s.d.”) of less than or equal to about 2 Å when superimposed, using backbone atoms (N, Cα, C and O), on the structure coordinates listed in Table 7 are considered to be identical with the structure coordinates listed in the Table when at least about 50% to 100% of the backbone atoms of the constituents of the WNV DIII-E16 Fab complex are included in the superposition.
Referring now to FIG. 9, the overall structure of the WNV DIII-E16 Fab complex shows that a convex pocket is formed by the variable regions of the heavy and light chains of E16 (VH and VL, respectively) which encompasses a discontinuous conformational epitope on WNV DII. All references by number to amino acids of the heavy and light chains of E16 are according to the Kabat numbering system, while references by number to amino acids of WNV DII correspond to the sequence identified with Genbank Accession No. AF196835. The buried surface area of the interaction (1548 Å2) is dominated by VH, which accounts for approximately 67% of the buried surface area. All six CDR loops approach within 4.2 Å of WNV DIII although CDR1 and CDR2 of the light chain are only minimally involved. The surface complementarity of the interaction is 0.763 (light chain 0.793 and heavy chain 0.742), consistent with previously published antibody-antigen interactions.
The residues in the VL that approach within 4.2 Å of WNV DIII can be separated into three segments, corresponding to CDR1, CDR2 and CDR3 (See Table 1, which lists all residues on WNV-DII that have any atoms within 4.2 Å of E16). The second column demonstrates the contacted residue in WNV-E16 and the distance of the closest approach. There are additional main chain contacts in residues A309, A331, A365, A389, L91, H96 and H99). CDR1 and CDR2 provide a few amino acid contacts to WNV DIII, using Ser30L and Trp50L, respectively. The VL CDR3 interaction is more extensive, utilizing amino acid residues 91-94. In contrast, the heavy chain dominates the interaction with WNV DIII (See Table 1). CDR1 contracts DIII using residues Tyr27H, Thr28H, Asp31H, Tyr32H and Trp33H. CDR2 makes connects with residues Leu52H, Arg56H and Arg58H. Interestingly, CDR3 has significant interactions with WNV DIII accounting for  ̃33% of the buried interface, with contacts at residues 94 through 100. As is commonly observed in antibody interactions with antigen, many of the contacting residues have aromatic side chains. For example, of 21 residues that contact WNV DIII, 6 are aromatic (2 Trp and 4 Tyr).







TABLE 1







Contact residues of E16 Fab and WNV DIII












WNV-DIII residue
E16 Residue
Chain
Distance
















302
56
H
3.54



306
33
H
2.61



306
52
H
3.57



307
33
H
3.80



307
100
H
4.02



307
95
H
2.49



307
96
H
2.87



308
98
H
3.84



309
98
H
3.84



330
50
L
3.61



330
98
H
4.03



330
99
H
3.42



331
100
H
3.95



332
94
L
3.76



332
58
H
2.77



332
100
H
2.89



332
91
L
3.26



333
33
H
4.09



333
56
H
3.12



333
58
H
4.19



365
93
L
2.98



366
92
L
3.47



366
93
L
4.05



367
50
L
3.86



368
30
L
2.59



368
50
L
3.70



389
31
H
3.08



390
27
H
3.83



390
28
H
2.59



390
31
H
3.96



390
32
H
2.47



390
94
H
4.12



391
31
H
3.00



391
32
H
3.74



391
97
H
2.79










Further, crystallographic analysis revealed that WNV DIII forms a β barrel structure formed from seven anti-parallel β-strands in two beta sheets (See FIG. 9), consistent with previous X-ray and NMR studies of other flavivirus E proteins. See Modis et al., 2003, Proc. Natl. Acad. Sci. USA 100:6986-91; Rey et al., 1995, Nature 375:291-8; Volk et al., 2004, J. Biol. Chem. 279:38755-38761; and Yu et al., 2004, Biochemistry 43:9168-76. One anti-parallel beta sheet consists of beta strands β1 (F309-D317), β2 (V323-Y329), β4 (D348-P351) and β5 (A369-E376), arranged in that order. The second anti-parallel beta sheet is formed from strands β3 (1340-S345), β6 (G380-V386) and β7 (1393-K399) arranged with β6 as the center. Based on antibody epitope mapping studies, amino acids that appear to define antigenic differences between WNV and other flaviviruses are located near the N-terminus of WNV DIII, and occupy a large proportion of the binding interface (see FIG. 10).
Based on the co-crystal structure, WNV DIII provides a nonlinear epitope for binding of the neutralizing E16 Fab. Four different loops of WNV DIII provide close contacts with the antibody, including residues 302 to 308, 330 to 333, 366 to 368 and 389 to 391. Because the contact area of the four loops that E16 Fab binds is not appreciably altered from that of unbound dengue and tick-borne encephalitis virus DIII (see Modis et al., 2003, Proc. Natl. Sci. USA 100:6986-91; Rey et al., 1995, Nature 375:291-8), the Fab does not appears to induce significant conformational change in WNV DIII. The structure of E16 Fab was also modeled onto the structure of the virion. The face of DIII that binds to E16 Fab is exposed to solvent and not sterically blocked by other envelope proteins in the virion. The Fab binding face points at the outer ring of the five-fold axis, in addition to other sites in the virion.
The interaction between the Fab and DIII is dominated by hydrogen bonding, with only a limited number of hydrophobic contacts. Of the 7 total hydrophobic contacts, Trp50L accounts for all but one of these (Ser30L-Asn368A), as it interacts with Thr330A, Asn368A and Ala367A (see Table 2). In comparison, there are 16 hydrogen bonds directly connecting the antibody to the antigen (see Table 3). Despite the large number of hydrogen bonds, no salt bridges are formed between DIII and E16. The hydrogen bonds generate an elaborate network, with a number of critical residues coordinating multiple bonds between DIII and E16. Notably, six of the hydrogen bonds involve main-chain interactions on WNV DIII. Such interactions may be invariant with respect to sequence and side-chain and thus, could explain the broad reactivity of this antibody with divergent lineage I and II strains. For example, Tyr92L and Ser97H contact WNV DIII through main-chain to main-chain interactions. In contrast, Thr93L, Asp31H, Arg56H, and Asp100H use their side chains to bond to main chain atoms in WNV DIII.







TABLE 2







Hydrophobic Contacts Between E16 and domain III












Atom 1

Atom 2
Distance



















ASN
A368
CB
SER
L30
CB
3.99



THR
A330
CA
TRP
L50
CZ2
3.66



THR
A330
CB
TRP
L50
CZ2
3.62



THR
A330
CG2
TRP
L50
CZ2
3.85



ASN
A368
CB
TRP
L50
CZ3
3.85



ASN
A368
CG
TRP
L50
CZ3
3.76



ALA
A367
CB
TRP
L50
CH2
3.87

















TABLE 3







Hydrogen Bond Contacts Between E16 and domain III












Donor

Acceptor
Distance



















SER
L30
OG
ASN
A368
OD1
2.59



ALA
A367
N
TYR
L92
O
2.82



THR
L93
OG1
ALA
A365
O
2.98



THR
H28
N
GLU
A390
OE2
2.61



THR
H28
OG1
GLU
A390
OE2
2.59



GLU
A390
N
ASP
H31
OD2
3.01



GLN
A391
NE2
ASP
H31
O
3.05



TYR
H32
OH
GLU
A390
OE1
2.48



TRP
H33
NE1
SER
A306
OG
2.62



ARG
H56
NH1
ASP
A333
O
2.91



ARG
H58
NH1
THR
A332
OG1
2.78



LYS
A307
NZ
SER
H95
OG
2.50



LYS
A307
NZ
ALA
H96
O
2.87



SER
H97
OG
GLN
A391
OE1
2.80



ALA
A308
N
SER
H97
O
2.79



THR
A332
N
ASP
H100
OD1
3.27



Wat
 24
OH2
SER
A306
O
2.74



Wat
 24
OH2
Wat
 7
OH2
2.63



Wat
 24
OH2
ASP
H31
O
2.88



LYS
A307
NZ
Wat
 7
OH2
2.92



TRP
H33
N
Wat
 7
OH2
2.77



Wat
 7
OH2
SER
H95
O
2.83



TYR
H98
OH
Wat
243
OH2
3.00



Wat
243
OH2
PHE
A309
O
2.71



Wat
 14
OH2
THR
A330
O
2.80



HIS
L91
ND1
Wat
 14
OH2
2.90



Wat
 14
OH2
HIS
L91
O
2.63



ASP
H100
N
Wat
 14
OH2
2.81



Wat
160
OH2
THR
A332
O
2.96



Wat
160
OH2
ALA
A365
O
2.83



THR
L93
OG1
Wat
160
OH2
2.96



THR
L94
N
Wat
160
OH2
3.08



THR
L94
OG1
Wat
160
OH2
3.07










Ten of the hydrogen bonds interact with side chains on WNV DIII. Lys307A uses its amine hydrogen to form hydrogen bonds with Ala96H and Ser95H, in addition to water-7. Glu390A is coordinated by Thr28H and Asp31H, and Gln391A contacts Asp31H and Ser97H. WNV DIII residues that contact multiple residues in E16 are likely to be required for high affinity antibody binding.
In addition to direct interactions between E16 and WNV DIII, numerous hydrogen bond networks are formed through intervening water molecules present at the antibody-antigen interface. All but one of the water molecules involve main chain atoms in WNV DIII. Lys307A is the only side chain that connects to the antibody by hydrogen bonding to Ser95H and Trp33H through water-7. Waters-14, -24, -160, and -243 all hydrogen bond with the main chain of DIII in addition to contacting multiple residues in E16. Although most of the water molecules are peripheral, waters-7 and 14 are found within the binding pocket.
5.6 Structure Coordinates
The atomic structure coordinates can be used in molecular modeling and design, as described more fully below. The present invention encompasses the structure coordinates and other information, e.g., amino acid sequence, connectivity tables, vector-based representations, temperature factors, etc., used to generate the three-dimensional structure of the polypeptide for use in the software programs described below and other software programs.
The invention encompasses machine-readable media embedded with the three-dimensional structure of the model described herein, or with portions thereof. As used herein, “machine-readable medium” refers to any medium that can be read and accessed directly by a computer or scanner. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc storage medium and magnetic tape; optical storage media such as optical discs or CD-ROM; electrical storage media such as RAM or ROM; and hybrids of these categories such as magnetic/optical storage media. Such media further include paper on which is recorded a representation of the atomic structure coordinates, e.g., Cartesian coordinates, that can be read by a scanning device and converted into a three-dimensional structure with an OCR.
A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon the atomic structure coordinates of the invention or portions thereof and/or X-ray diffraction data. The choice of the data storage structure will generally be based on the means chosen to access the stored information. In addition, a variety of data processor programs and formats can be used to store the sequence and X-ray data information on a computer readable medium. Such formats include, but are not limited to, Protein Data Bank (“PDB”) format (Research Collaboratory for Structural Bioinformatics; Cambridge Crystallographic Data Centre format; Structure-data (“SD”) file format (MDL Information Systems, Inc.; Dalby et al., 1992, J. Chem. Inf. Comp. Sci. 32:244-255), and line-notation, e.g., as used in SMILES (Weininger, 1988, J. Chem. Inf. Comp. Sci. 28:31-36). Methods of converting between various formats read by different computer software will be readily apparent to those of skill in the art, e.g., BABEL (v. 1.06, Walters & Stahl, ©1992, 1993, 1994). All format representations of the polypeptide coordinates described herein, or portions thereof, are contemplated by the present invention. By providing computer readable medium having stored thereon the atomic coordinates of the invention, one of skill in the art can routinely access the atomic coordinates of the invention, or portions thereof, and related information for use in modeling and design programs, described in detail below.
While Cartesian coordinates are important and convenient representations of the three-dimensional structure of a polypeptide, those of skill in the art will readily recognize that other representations of the structure are also useful. Therefore, the three-dimensional structure of a polypeptide, as discussed herein, includes not only the Cartesian coordinate representation, but also all alternative representations of the three-dimensional distribution of atoms. For example, atomic coordinates may be represented as a Z-matrix, wherein a first atom of the protein is chosen, a second atom is placed at a defined distance from the first atom, a third atom is placed at a defined distance from the second atom so that it makes a defined angle with the first atom. Each subsequent atom is placed at a defined distance from a previously placed atom with a specified angle with respect to the third atom, and at a specified torsion angle with respect to a fourth atom. Atomic coordinates may also be represented as a Patterson function, wherein all interatomic vectors are drawn and are then placed with their tails at the origin. This representation is particularly useful for locating heavy atoms in a unit cell. In addition, atomic coordinates may be represented as a series of vectors having magnitude and direction and drawn from a chosen origin to each atom in the polypeptide structure. Furthermore, the positions of atoms in a three-dimensional structure may be represented as fractions of the unit cell (fractional coordinates), or in spherical polar coordinates.
Additional information, such as thermal parameters, which measure the motion of each atom in the structure, chain identifiers, which identify the particular chain of a multi-chain protein in which an atom is located, and connectivity information, which indicates to which atoms a particular atom is bonded, is also useful for representing a three-dimensional molecular structure.
5.7 Uses of the Automic Structure Coordinates
Structure information, typically in the form of the atomic structure coordinates, can be used in a variety of computational or computer-based methods to, for example, design, screen for and/or identify compounds that bind the crystallized polypeptide or a portion or fragment thereof, to intelligently design mutants that have altered biological properties, to intelligently design and/or modify antibodies that have desirable binding characteristics, and the like.
In one embodiment, the crystals and structure coordinates obtained therefrom are useful for identifying and/or designing compounds that bind WNV DIII as an approach towards developing new therapeutic agents. For example, a high resolution X-ray structure will often show the locations of ordered solvent molecules around the protein, and in particular at or near putative binding sites on the protein. This information can then be used to design molecules that bind these sites, the compounds synthesized and tested for binding in biological assays. See Travis, 1993, Science 262:1374. Such compounds may be useful, for example, to inhibit binding of WNV E protein to its cognate receptor or to induce production of antibodies or other immune responses that can protect against WNV infection.
In another embodiment, the structure is probed with a plurality of molecules to determine their ability to bind to the WNV DIII alone or in complex with a monoclonal antibody such as E16 at various sites. Such compounds can be used as targets or leads in medicinal chemistry efforts to identify, for example, inhibitors of potential therapeutic importance or molecules that enhance antibody binding and/or neutralization of WNV DIII.
In yet another embodiment, the structure is probed with a plurality of molecules to determine their ability to bind to the WNV DIII binding site of E16. Such compounds can be used, for example, as targets or leads in medicinal chemistry efforts to identify, for example, compounds that can be used to induce an immune response that can provide prophylactic and/or therapeutic benefit against WNV infection.
In yet another embodiment, the structure can be used to computationally screen small molecule data bases for chemical entities or compounds that can bind in whole, or in part, to WNV DIII. In this screening, the quality of fit of such entities or compounds to the binding site may be judged either by shape complementarity or by estimated interaction energy. See Meng et al., 1992, J. Comp. Chem. 13:505-524.
The design of compounds that bind to WNV DIII, inhibit the interaction of WNV DIII with its cognate receptor, or inhibit WNV infection, according to this invention generally involves consideration of two factors. First, the compound should be capable of physically and structurally associating with WNV DIII. This association can be covalent or non-covalent. For example, covalent interactions may be important for designing irreversible inhibitors of WNV DIII. Non-covalent molecular interactions important in the association of WNV DIII with its substrate include hydrogen bonding, ionic interactions and van der Waals and hydrophobic interactions. Second, the compound should be able to assume a conformation that allows it to associate with WNV DIII. Although certain portions of the compound will not directly participate in this association with WNV DIII, those portions may still influence the overall conformation of the molecule. This, in turn, may impact potency. Such conformational requirements include the overall three-dimensional structure and orientation of the chemical group or compound in relation to all or a portion of the binding site, or the spacing between functional groups of a compound comprising several chemical groups that directly interact with WNV DIII.
The potential inhibitory or binding effect of a chemical compound on WNV DIII binding may be analyzed prior to its actual synthesis and testing by the use of computer modeling techniques. If the theoretical structure of the given compound suggests insufficient interaction and association between it and WNV DIII, synthesis and testing of the compound is unnecessary. However, if computer modeling indicates a strong interaction, the molecule may then be synthesized and tested for its ability to bind to WNV DIII and inhibit its binding activity. In this manner, synthesis of ineffective compounds may be avoided.
An inhibitory or other binding compound of WNV DIII may be computationally evaluated and designed by means of a series of steps in which chemical groups or fragments are screened and selected for their ability to associate with the individual binding pockets or other areas of WNV DIII. One skilled in the art may use one of several methods to screen chemical groups or fragments for their ability to associate with WNV DIII. This process may begin by visual inspection of, for example, the binding site on the computer screen based on the WNV DIII coordinates. Selected fragments or chemical groups may then be positioned in a variety of orientations, or docked, within an individual binding pocket of WNV DIII as described above. Docking may be accomplished using software such as QUANTA and SYBYL, followed by energy minimization and molecular dynamics with standard molecular mechanics force fields, such as CHARMM and AMBER.
These principles may also be used to design and evaluate compounds that can mimic WNV DIII. However, in designing and evaluating such compounds, the chemical groups or fragments are screened and selected for their ability to associate with the individual binding pockets of E16 that recognize WNV DIII. Preferably, the chemical groups or fragments together are able to associate with two, three, four, five, or six of the individual binding pockets that together recognize WNV DIII.
Specialized computer programs may also assist in the process of selecting fragments or chemical groups. These include:
1. GRID (Goodford, 1985, J. Med. Chem. 28:849-857). GRID is available from Oxford University, Oxford, UK;
2. MCSS (Miranker & Karplus, 1991, Proteins: Structure, Function and Genetics 11:29-34). MCSS is available from Molecular Simulations, Burlington, Mass.;
3. AUTODOCK (Goodsell & Olsen, 1990, Proteins: Structure, Function, and Genetics 8:195-202). AUTODOCK is available from Scripps Research Institute, La Jolla, Calif.; and
4. DOCK (Kuntz et al., 1982, J. Mol. Biol. 161:269-288). DOCK is available in several versions from University of California, San Francisco, Calif.
5. CombiDOCK (Sun et al., 1998, J. Comput. Aided Mol. Des. 12:597-604). CombiDOCK is available from Univeristy of California, San Francisco, Calif.).
6. FRED (available from OpenEye Scientific Software, Santa Fe, N. Mex.). Based on an original scientific perspective and efficient computational algorithms, FRED is an accurate and extremely fast docking program. With equivalent resolution settings, FRED will out-perform all known docking programs, typically examining about a dozen ligand conformers in a second. For each, FRED exhaustively examines all possible poses within the protein active site, filtering for shape complementarity and pharmacophoric features before scoring with more traditional functions.
7. ROCS (available from OpenEye Scientific Software, Santa Fe, N. Mex.). ROCS is a shape comparison program, based on the idea that molecules have similar shape if their volumes overlay well and any volume mismatch is a measure of dissimilarity. ROCS uses a smooth Gaussian function to represent the molecular volume, so it is possible to routinely minimize to the best global match. ROCS is capable of processing 600-800 comparisons each second, making it possible to search multi-conformer representations of corporate collections in a day on a single processor to find compounds with similar shape to a lead compound.
Once suitable chemical groups or fragments have been selected, they can be assembled into a single compound or inhibitor. Assembly may proceed by visual inspection of the relationship of the fragments to each other in the three-dimensional image displayed on a computer screen in relation to the structure coordinates of WNV DIII. This would be followed by manual model building using software such as QUANTA or SYBYL.
Useful programs to aid one of skill in the art in connecting the individual chemical groups or fragments include:
1. CAVEAT (Bartlett et al., 1989, “CAVEAT: A Program to Facilitate the Structure-Derived Design of Biologically Active Molecules,” In Molecular Recognition in Chemical and Biological Problems', Special Pub., Royal Chem. Soc. 78:182-196). CAVEAT is available from the University of California, Berkeley, Calif.;
2. 3D Database systems such as MACCS-3D (MDL Information Systems, San Leandro, Calif). This area is reviewed in Martin, 1992, J. Med. Chem. 35:2145-2154); and
3. HOOK (available from Molecular Simulations, Burlington, Mass.).
4. FILTER (available from OpenEye Scientific Software, Santa Fe, N. Mex.). FILTER is a molecular screening and selection tool that uses a combination of physical-property calculations and functional-group knowledge to assess compound collections. In selection mode, FILTER can be used to choose reagents appropriate for specific syntheses. In filter mode, it quickly removes compounds with undesirable elements, functional groups, or physical properties. FILTER is a command line utility that reads and writes many file formats including SDF, Mol, Mol2, SMILES, and OEBinary. Filter is supported on Linux, Windows and ilux; other platforms are available upon request.
5. SMACK (available from OpenEye Scientific Software, Santa Fe, N. Mex.). SMACK converts and optimizes molecular database queries. SMACK can quickly convert from substructure and reaction queries expressed in MDL file format to SMARTS strings. Additionally, SMACK will automatically optimize each resulting query for pattern-matching performance. Typical transformations simplify redundant atom and bond expressions and reorder atoms for faster matching against medicinal/organic chemistry databases.
Instead of proceeding to build a WNV DIII binding inhibitor in a step-wise fashion one fragment or chemical group at a time, as described above, WNV DIII binding compounds may be designed as a whole or “de novo” using either an empty WNV DIII binding site or optionally including some portion(s) of a known inhibitor(s). These methods include:
1. LUDI (Bohm, 1992, J. Comp. Aid. Molec. Design 6:61-78). LUDI is available from Molecular Simulations, Inc., San Diego, Calif.;
2. LEGEND (Nishibata & Itai, 1991, Tetrahedron 47:8985). LEGEND is available from Molecular Simulations, Burlington, Mass.; and
3. LeapFrog (available from Tripos, Inc., St. Louis, Mo.).
4. WABE (available from OpenEye Scientific Software, Santa Fe, N. Mex.). WABE is a de similis design program, as opposed to de novo design. Using a graph-invariant replacement algorithm, it quickly generates large numbers of isosteres to a lead molecule. The method resembles the process of chemical substitution, e.g. carboxylate to amnide or amidine, used in medicinal chemistry to create molecules having the same physical shape but varying in electrostatics. The similarities between analogs eliminate computationally demanding terms, so WABE can also quickly rank them by electrostatic similarity to a known binder or by protein-ligand binding. WABE is useful for exploring the chemical space around a lead compound to elucidate candidates with improved drug profiles or novel scaffolds to avoid patent coverage.
Other modeling and simulation computer programs include, but are not restricted to the following:
1. AMBER (available from University of California, San Francisco). AMBER (Assisted Model Building with Energy Refinement) is a molecular dynamics and energy minimization program.
2. CHARMM (available from Harvard University). CHARMM (Chemistry at HARvard Macromolecular Mechanics) is a program for macromolecular simulations, including energy minimization, molecular dynamics and Monte Carlo simulations.
3 Disulfide by Design (available from Wayne State University). Disulfide by Design is an application for the rational design of disulfide bonds in proteins and for exporting mutant PDB files containing the modeled disulfides for visualization in other molecular modeling software. For a given protein structural model, all residue pairs are rapidly assessed for proximity and geometry consistent with disulfide formation assuming the residues were mutated to cysteines. The output displays residue pairs meeting the appropriate criteria. The input model will typically be a PDB structure for the protein of interest; however, structures developed through homology modeling may also be used. Engineered disulfides have proven useful for increasing the stability of proteins and to assist the investigation of protein dynamics and interactions.
4. FTDOCK (available from the Biomolecular Modelling Laboratory of Cancer Research UK). FTDOCK is a program for carrying out rigid-body docking between biomolecules.
5. GROMOS (available from Laboratory of Physical Chemistry; ETH Honggerberg, HC). GROMOS is a general purpose molecular dynamics computer simulation package for the study of biomolecular systems.
6. GROMACS (an open-source tool freely available on the internet). GROMACS is a complete modeling package for proteins, membrane systems and more, including fast molecular dynamics, normal mode analysis, essential dynamics analysis and many trajectory analysis utilities.
7. ICM (Molsoft LLC, San Diego, Calif.). ICM from Molsoft provides programs and modules for applications including for structure analysis, modeling, docking, homology modeling and virtual ligand screening.
8. JACKAL (available from Columbia University). JACKAL is a suite of tools for model building, structure prediction and refinement, reconstruction, and minimization; for SGI, Linux, and Sun Solaris
9. LOOPP (available from Cornell University). LOOP (Linear Optimization of Protein Potentials) is available form Cornell Theory Center program for potential optimization and alignments of sequences and structures.
10. MAMMOTH (available from New York University). MAMMOTH (MAtching Molecular Models Obtained from Theory) is a program for automated pairwise and multiple structural alignments; for SGI, Linux, and Sun Solaris.
11. MCCE (available from the City College of New York). The MCCE (Multi-Conformation Continuum Electrostatics) software calculates theoretical pKas of residues in proteins and provides the modulating factors of pKas based on the structure in PDB format.
12. MidasPlus (available from University of California, San Francisco) MidasPlus is a program for displaying, manipulating and analysing macromolecules.
13. MODELLER (available from University of California, San Francisco). MODELLER is a program for automated protein homology modeling.
14. MOIL (available from Cornell University). MOIL is another program from the Cornell Theory Center package for molecular dynamics simulation of biological molecules.
15. NAMD (available from the University of Illinois at Urbana-Champaign). NAMD is a parallel object-oriented molecular dynamics simulation program.
16. WAM (available from the University of Bath). WAM (Web Antibody Modeling) provides a server for automated structure modeling from antibody Fv sequences.
17. 123D (Ceres Inc., Malibu, Calif.). 123D is a program which threads a sequence through a set of structures using substitution matrix, secondary structure prediction and contact capacity potential.
Additional molecular modeling techniques may also be employed in accordance with this invention. See, e.g., Cohen et al., 1990, J. Med. Chem. 33:883-894. See also Navia & Murcko, 1992, Cur. Op. Struct. Biol. 2:202-210.
Once a compound has been designed or selected by the above methods, the efficiency with which that compound may bind to WNV DIII may be tested and optimized by computational evaluation. For example, a compound that has been designed or selected to function as a WNV DIII binding inhibitor should also preferably occupy a volume not overlapping the volume occupied by the binding site residues when the native receptor is bound. An effective WNV DIII inhibitor preferably demonstrates a relatively small difference in energy between its bound and free states (i.e., it should have a small deformation energy of binding). Thus, the most efficient WNV DIII binding inhibitors should preferably be designed with a deformation energy of binding of not greater than about 10 kcal/mol, preferably, not greater than 7 kcal/mol. WNV DIII binding inhibitors may interact with the protein in more than one conformation that is similar in overall binding energy. In those cases, the deformation energy of binding is taken to be the difference between the energy of the free compound and the average energy of the conformations observed when the inhibitor binds to the enzyme.
A compound selected or designed for binding to WNV DIII may be further computationally optimized so that in its bound state it would preferably lack repulsive electrostatic interaction with the target protein. Such non-complementary electrostatic interactions include repulsive charge-charge, dipole-dipole and charge-dipole interactions. Specifically, the sum of all electrostatic interactions between the inhibitor and the protein when the inhibitor is bound to it preferably make a neutral or favorable contribution to the enthalpy of binding.
Specific computer software is available in the art to evaluate compound deformation energy and electrostatic interaction. Examples of programs designed for such uses include: Gaussian 92, revision C (Frisch, Gaussian, Inc., Pittsburgh, Pa. (01992); AMBER, version 4.0 (Kollman, University of California at San Francisco, ©1994); QUANTA/CHARMM (Molecular Simulations, Inc., Burlington, Mass., ©1994); and Insight II/Discover (Biosym Technologies Inc., San Diego, Calif., (1994). These programs may be implemented, for instance, using a computer workstation, as are well-known in the art. Other hardware systems and software packages will be known to those skilled in the art.
Once a WNV DIII-binding compound has been optimally selected or designed, as described above, substitutions may then be made in some of its atoms or chemical groups in order to improve or modify its binding properties. Generally, initial substitutions are conservative, i.e., the replacement group will have approximately the same size, shape, hydrophobicity and charge as the original group. One of skill in the art will understand that substitutions known in the art to alter conformation should be avoided. Such altered chemical compounds may then be analyzed for efficiency of binding to WNV DIII by the same computer methods described in detail above.
Because WNV DIII-E16 Fab complex may crystallize in more than one form, the structure coordinates of WNV DIII-E16 Fab complex, or portions thereof, are particularly useful to solve the structure of those other crystal forms of WNV DIII-E16 Fab complex. They may also be used to solve the structure of WNV DIII-E16 Fab complex mutants, WNV DIII-E16 Fab poly-complex that is complexed with one or more additional molecules, or of the crystalline form of any other protein with significant amino acid sequence homology to any functional domain of WNV DIII and/or E16 Fab.
One method that may be employed for this purpose is molecular replacement. In this method, the unknown crystal structure, whether it is another crystal form of WNV DIII-E16 Fab complex, a mutant WNV DIII-E16 Fab complex, a WNV DIII-mutant E16 Fab complex, or a WNV DIII-E16 Fab poly-complex that is complexed with one or more additional molecules, or the crystal of some other protein with significant amino acid sequence homology to any functional domain of WNV DIII and/or E16 Fab, may be determined using phase information from the WNV DIII-E16 Fab complex structure coordinates. This method may provide an accurate three-dimensional structure for the unknown protein in the new crystal more quickly and efficiently than attempting to determine such information ab initio. In addition, in accordance with this invention, WNV DIII-E16 Fab complex mutants may be crystallized in complex with known WNV DIII binding inhibitors. The crystal structures of a series of such complexes may then be solved by molecular replacement and compared with that of wild-type WNV DIII. Potential sites for modification within the various binding sites of the protein may thus be identified. This information provides an additional tool for determining the most efficient binding interactions, for example, increased hydrophobic interactions, between WNV DIII and a chemical group, compound, or monoclonal antibody such as E16.
If an unknown crystal form has the same space group as and similar cell dimensions to the known orthorhombic crystal form, then the phases derived from the known crystal form can be directly applied to the unknown crystal form, and in turn, an electron density map for the unknown crystal form can be calculated. Difference electron density maps can then be used to examine the differences between the unknown crystal form and the known crystal form. A difference electron density map is a subtraction of one electron density map, e.g., that derived from the known crystal form, from another electron density map, e.g., that derived from the unknown crystal form. Therefore, all similar features of the two electron density maps are eliminated in the subtraction and only the differences between the two structures remain. For example, if the unknown crystal form is of a WNV DIII-E16 Fab complex in complex with one or more additional molecules, then a difference electron density map between this map and the map derived from the WNV DIII-E16 Fab complex crystal will ideally show only the electron density of the ligand. Similarly, if amino acid side chains have different conformations in the two crystal forms, then those differences will be highlighted by peaks (positive electron density) and valleys (negative electron density) in the difference electron density map, making the differences between the two crystal forms easy to detect. However, if the space groups and/or cell dimensions of the two crystal forms are different, then this approach will not work and molecular replacement can be used in order to derive phases for the unknown crystal form.
All of the complexes referred to above may be studied using well-known X-ray diffraction techniques and may be refined versus 50 Å to 1.5 Å or greater resolution X-ray data to an R value of about 0.20 or less using computer software, such as CNS (Yale University, (c) 1992, distributed by Molecular Simulations, Inc.). See, e.g., Blundel et al., 1976, Protein Crystallography, Academic Press.; Methods in Enzymology, vols. 114 & 115, Wyckoff et al., eds., Academic Press, 1985. This information may thus be used to optimize known classes of WNV DIII binding inhibitors, and more importantly, to design and synthesize novel classes of WNV DIII binding inhibitors.
The structure coordinates of mutant WNV DIII-E16 Fab complex and/or WNV DIII-mutant E16 Fab complex will also facilitate the identification of related proteins or enzymes analogous to WNV DIII and/or E16 Fab in function, structure or both, thereby further leading to novel therapeutic modes for treating or preventing WNV or other flavivirus-mediated diseases.
Subsets of the atomic structure coordinates can be used in any of the above methods. Particularly useful subsets of the coordinates include, but are not limited to, coordinates of single domains, coordinates of residues lining an active site, coordinates of residues that participate in important protein-protein contacts at an interface, and Ca coordinates. For example, the coordinates of one domain of a protein that contains the active site may be used to design inhibitors that bind to that site, even though the protein is fully described by a larger set of atomic coordinates. Therefore, a set of atomic coordinates that define the entire polypeptide chain, although useful for many applications, do not necessarily need to be used for the methods described herein.
5.8 Methods of Identifying Neutralizing
Epitopes of Flaviviral Envelope Proteins
In yet another aspect, the invention provides a method for identifying a neutralizing epitope of a domain III of a flaviviral envelope protein. In certain embodiments, the method comprises comparing an amino acid sequence of the domain III of the flaviviral envelope protein to an amino acid sequence of a domain III of a WNV E protein, and identifying the amino acids of the domain III of the flaviviral envelope protein corresponding to a neutralizing epitope of the domain III of the WNV E protein, thereby identifying the neutralizing epitope of the flaviviral envelope protein. The methods can be performed with any flaviviral envelope protein known to one of skill in the art without limitation. For example, the methods can be performed with flaviviral envelope protein from Japanese Encephalitus Virus, Dengue Virus, Yellow Fever Virus, St. Louis Encephalitus Virus, Murray Valley Encephalitus Virus, Rocio Virus, Tick-borne Encephalitus Virus, Kyasanur Forest Disease Virus, Omsk Hemorrhagic Fever Virus, Montana myotis leukoencephalitis virus, Modoc virus, Powssan Virus, and Kunjin Virus.
The amino acid sequences of these envelope proteins, and references describing the determination of these amino acid sequences are described in Table 4, below










TABLE 4





Virus
Accession Number
For
Reference







Japanese Encephalitus Virus
NP_775666
Envelope protein
Sumiyoshi et al., 1987, Virology 161(2), 497–510


Dengue Virus
NP_739583
E protein
Hahn et al., 1988, Virology 162(2), 167–180


Yellow Fever Virus
NP_740305
Envelope protein
Rice et al., 1985, Science 229(4715), 726–733


St. Louis Encephalitus Virus
AAF22698 to
Envelope protein
Charrel et al., 1999, J. Clin. Microbiol. 37(6),



AAF22719

1936–1940


Murray Valley Encephalitus
NP_722531
Envelope protein
Hurrelbrink et al., 1999, J. Gen. Virol. 80(pt 12),


Virus


3115–3125


Rocio Virus
AAK91596
Envelope protein
Gaunt et al., 2001, J. Gen. Virol. 82(pt 8),





1867–1876


Tick-borne Encephalitus Virus
NP_775503
Envelope protein
Wallner et al., 1995, Virology 213(1), 169–178


Kyasanur Forest Disease Virus
X74111
Virus DNA sequence
Venugopal et al., 1994, J. Gen. Virol. 75 (pt1),




for core, membrane
227–232




& envelope proteins


Omsk Hemorrhagic Fever Virus
NP_932085
Envelope protein
Lin et al., 2003, Virology 313(1), 81–90


Montana myotis
NP_740277
Envelope protein
Charlier et al., 2002, J. Gen. Virol. 80(pt 8),


leukoencephalitis virus


1875–1885


Modoc virus
NP_740260
Envelope protein
Leyssen et al., 2002, Thesis, Department of





Microbiology, Rega Institute for Medical





Research, Leuven, Belguim


Powssan Virus
NP_775516
Envelope protein
Mandl et al., 1993, Virology 194(1), 173–184


Kunjin Virus
P14335
Amino acid sequence
Coia et al., 1988, J. Gen. Virol. 69: 1–21




of viral polyprotein;




envelope protein




defined by amino




acids 291–791









In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 330-333 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. Exemplary regions corresponding to portions of the neutralizing epitope of WNV DIII are presented in FIG. 15.
Any suitable method known to one of skill in the art can be used to compare sequences of envelope proteins in the methods of the invention. Conveniently, such comparisons can be performed using one of many suitable computer-implemented algorithms known to the art. These algorithms generally identify regions of greatest homology and identity between two or more related amino acid and/or nucleotide sequences and thus can be used to identify sequences in flaviviral envelope proteins that correspond to sequences that make up, in whole or in part, a WNV neutralizing epitope as described herein.
Exemplary algorithms which can be used to determine identity and/or homology between two sequences include, but are not limited to, the suite of BLAST programs, e.g., BLASTN, BLASTX, and TBLASTX, BLASTP and TBLASTN, publicly available on the Internet at the NCBI website. See also Altschul et al., 1990, J. Mol. Biol. 215:403-10 (with special reference to the published default setting, i.e., parameters w=4, t=17) and Altschul et al., 1997, Nucleic Acids Res., 25:3389-3402. Sequence searches are typically carried out using the BLASTP program when evaluating a given amino acid sequence relative to amino acid sequences in the GenBank Protein Sequences and other public databases. The BLASTX program is preferred for searching nucleic acid sequences that have been translated in all reading frames against amino acid sequences in the GenBank Protein Sequences and other public databases. Both BLASTP and BLASTX are run using default parameters of an open gap penalty of 11.0, and an extended gap penalty of 1.0, and utilize the BLOSUM-62 matrix. See id.
Alignments of selected sequences in order to determine homology and/or identity between two or more sequences, can also be performed using for example, the CLUSTAL-W program in MacVector version 6.5, operated with default parameters, including an open gap penalty of 10.0, an extended gap penalty of 0.1, and a BLOSUM 30 similarity matrix.
The present invention is also intended to encompass immunogens comprising neutralizing epitopes from the flaviviruses described above, antibodies that specifically bind such neutralizing epitopes, methods of inducing immune responses against such epitopes, methods of treating or preventing infection with the flaviviruses, and the like. One of skill in the art can, after identifying such neutralizing epitopes as described above, construct such immunogens, make such antibodies, and perform such methods by adapting the invention described herein to use the neutralizing epitope identified above.
5.9 Methods of Making High Affinity Antibodies to WNV DIII
In yet another aspect, the present invention provides methods of making antibodies that bind WNV DIII with high affinity. In general, the methods rely on use of the three-dimensional structure of WNV DIII-E16 Fab complex to identify one or more amino acid residues present in the Fab fragment that can be altered in order to increase the affinity of E16 for WNV DIII. Thus, in certain embodiments, the methods comprise mutating an amino acid of the antibody which when mutated is predicted to increase the affinity with which the antibody binds the domain III of WNV E protein.
In certain embodiments, the amino acid that is mutated contacts the domain III of WNV E protein more closely than the wild-type amino acid. In certain embodiments, the mutated amino acid excludes a water molecule from a hydrophobic region of the domain III that is not excluded by the wild-type amino acid. In certain embodiments, the mutated amino acid forms a hydrogen bond with the domain III that is not formed by the first amino acid. In certain embodiments, the mutated amino acid is an amino acid selected from Table 1. In certain embodiments, the mutated amino acid is an amino acid selected from Table 2. In certain embodiments, the mutated amino acid is an amino acid selected from Table 3.
In certain embodiments, more than one amino acid of the antibody can be mutated. Thus, in certain embodiments, one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, twenty, twenty-five, thirty, forty, fifty, or more amino acids can be mutated. In certain embodiments, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, twenty, twenty-five, or twenty-nine amino acids selected from Table 1 are mutated. In certain embodiments, two, three, four, five, six, or seven amino acids selected from Table 2 are mutated. In certain embodiments, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, twenty, twenty-five, or thirty amino acids selected from Table 3 are mutated.
In still another aspect, the invention provides methods of making antibodies that bind flaviviral envelope proteins with high affinity. Using the three-dimensional structural coordinates provided in the present application and structural information about flaviviral envelope proteins available to one skilled in the art, one or more amino acid residues that affect the affinity of an antibody, or fragment thereof, for a flaviviral envelope protein can be identified. Such amino acids can be mutated to increase or decrease the affinity of the antibody for the envelope protein. Thus, the methods of the invention can also be used to make antibodies that bind with high affinity an envelope protein from a flavivirus, including, but not limited to, WNV, Japanese encephalitis virus, Kunjin virus, Murray Valley encephalitis virus, Dengue virus, Montana myotis leukoencephalitis virus, Modoc virus, and Yellow Fever virus.
Exemplary sources available to one skilled in the art for structural information regarding envelope proteins from other flaviviruses may be found, for example, in Beasley et al., 2002, J. Virol. 76:13097-13100; Mukhopadhyay et al., 2003, Science 302:248; Rey et al., 1995, Nature 375:291-8; Roehrig et al., 2001, Ann. N.Y. Acad. Sci. 951:286-297; Zhang et al., 2003, EMBO J 22:2604-13; and Zhang et al., 2003, Nat. Struct. Biol. 10:907-912; Kuhn et al., 2002, Cell 108:717-25; Modis et al., 2003, Proc. Natl. Acad. Sci. USA 100:6986-91; Beasley and Aaskov, 2001, Virology 279:447-58; Cecilia and Gould, 1991, Virology 181:70-7; Crill and Roehrig, 2001, J. Virol. 75:7769-73; Lin et al., 1994, Virology 202:885-90; Roehrig et al., 1983, Virology 128:118-26; Schlesinger et al., 1996, J. Gen. Virol. 77:1277-85; Seifetal., 1995, Vaccine 13:1515-21; Volketal., 2004, J. Biol. Chem. 279:38755-38761; Wu et al., 1997, Virus. Res. 51:173-81; and Wu et al., 2003, J. Biol. Chem. 278:46007-46013.
Standard techniques known to those skilled in the art can be used to introduce mutations in the nucleotide sequence encoding an antibody, or fragment thereof, including, e.g., site-directed mutagenesis and PCR-mediated mutagenesis, which results in amino acid substitutions. In certain embodiments, the E16 derivatives include less than 15 amino acid substitutions, less than 10 amino acid substitutions, less than 5 amino acid substitutions, less than 4 amino acid substitutions, less than 3 amino acid substitutions, or less than 2 amino acid substitutions relative to the original antibody or fragment thereof. In a preferred embodiment, the derivatives have conservative amino acid substitutions made at one or more amino acid residues wherein the substitutions are predicted to increase the affinity of the antibody for WNV DIII. In an equally preferred embodiment, the derivatives have conservative amino acid substitutions made at one or more amino acid residues wherein the substitutions are predicted to increase the affinity of the antibody for WNV DIII
The antibodies thus made may be characterized for specific binding to a WNV DIII, or any other flaviviral envelope protein, using any immunological or biochemical based method known in the art for characterizing including quantitating, the interaction of the antibody to a flaviviral envelope protein. Specific binding of an antibody to a flaviviral envelope protein may be determined, for example, using immunological or biochemical based methods including, but not limited to, an ELISA assay, surface plasmon resonance assays, immunoprecipitation assays, affinity chromatography, and equilibrium dialysis. Immunoassays which can be used to analyze immunospecific binding and cross-reactivity of the antibodies of the invention include, but are not limited to, competitive and non-competitive assay systems using techniques such as western blots, radioimmunoassays, ELISA assays, “sandwich” immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, and protein A immunoassays, to name but a few. Such assays are routine and well known in the art (see, e.g., Ausubel et al., eds, 2004, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, which is incorporated by reference herein in its entirety).
Antibodies that bind to a flaviviral envelope protein may also be assayed using any surface plasmon resonance based assays known in the art for characterizing the kinetic parameters of the interaction of the antibody with a flaviviral. Any SPR instrument commercially available including, but not limited to, BIAcore Instruments, available from Biacore AB (Uppsala, Sweden); IAsys instruments available from Affinity Sensors (Franklin, Mass.); IBIS system available from Windsor Scientific Limited (Berks, UK), SPR-CELLIA systems available from Nippon Laser and Electronics Lab (Hokkaido, Japan), and SPR Detector Spreeta available from Texas Instruments (Dallas, Tex.) can be used in the instant invention. For a review of SPR-based technology see Mullet et al., 2000, Methods 22: 77-91; Dong et al., 2002, Review in Mol. Biotech., 82: 303-23; Fivash et al., 1998, Current Opinion in Biotechnology 9: 97-101; Rich et al., 2000, Current Opinion in Biotechnology 11: 54-61; all of which are incorporated herein by reference in their entirety. Additionally, any of the SPR instruments and SPR based methods for measuring protein-protein interactions described in U.S. Pat. Nos. 6,373,577; 6,289,286; 5,322,798; 5,341,215; and 6,268,125 are contemplated in the methods of the invention, all of which are incorporated herein by reference in their entirety.
The invention also encompasses characterization of the antibodies produced by the methods of the invention using certain characterization assays for identifying the function of the antibodies of the invention, particularly the activity to inhibit a flaviviral infection using in vitro and in vivo based assays. Further, the invention encompasses cell based and cell free assays for characterization or use of the antibodies. Additional methods of characterizing antibodies made according to a method of the invention are described below.
5.10 Antibodies Binding a Neutralizing Epitope of WNV DIII
In yet another aspect, the invention provides antibodies, or fragments thereof, that specifically bind a neutralizing epitope of WNV DIII as described herein. In certain embodiments, the antibody, or fragment thereof, is a purified antibody, or fragment thereof. In certain embodiments, the antibody, or fragment thereof, is a monoclonal antibody, or fragment thereof. In certain embodiments, the antibody, or fragment thereof, is an antibody, or fragment thereof, purified from a polyclonal antibody preparation such that the antibody, or fragment thereof, is substantially isolated from antibodies, or fragments thereof, that bind WNV epitopes other than neutralizing epitopes. In certain embodiments, the antibody, or fragment thereof, is in the form of a composition that is purified to homogeneity. In certain embodiments, the antibody, or fragment thereof, is not 5H10, 3A3, 5C5, 7H2, 11C, 17C8, 10C5, 8B10, E1, E16, E24, E27, E33, E34, E40, E43, E47, E49, and/or E58, or any fragment thereof.
In certain embodiments, the antibody, or fragment thereof, binds the neutralizing epitope of domain III of WNV E protein with higher affinity than the antibody, or fragment thereof, binds an enhancing epitope of domain III of WNV E protein. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 330-333 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering.
In another aspect, the invention provides a pharmaceutical composition comprising an antibody, or fragment thereof, of the invention, as described herein, and a pharmaceutically acceptable excipient, diluent, vehicle, or carrier. In certain embodiments, the pharmaceutical composition comprises a monoclonal antibody. In certain embodiments, the pharmaceutical composition is a single unit dosage form. In certain embodiments, the pharmaceutical composition further comprises a stabilizer. In certain embodiments, the pharmaceutical composition is formulated for storage for at least about 6 months without loss of more than about 20% activity.
In another aspect, the invention provides a kit comprising a pharmaceutical composition of the invention. In certain embodiments, the antibody of the pharmaceutical composition is a monoclonal antibody. In certain embodiments, the pharmaceutical composition is in a single unit dosage form. In certain embodiments, the kit further comprises instructions directing administration of the pharmaceutical composition to a subject.
The antibodies or fragments thereof may be characterized in a variety of ways in addition to those described above. In particular, antibodies of the invention or fragments thereof may be assayed for the ability to immunospecifically bind to a WNV antigen. Such an assay may be performed in solution (e.g., Houghten, 1992, Bio/Techniques 13:412-421), on beads (Lam, 1991, Nature 354:82-84), on chips (Fodor, 1993, Nature 364:555-556), on bacteria (U.S. Pat. No. 5,223,409), on spores (U.S. Pat. Nos. 5,571,698; 5,403,484; and 5,223,409), on plasmids (Cull et al., 1992, Proc. Natl. Acad. Sci. USA 89:1865-1869) or on phage (Scott and Smith, 1990, Science 249:386-390; Devlin, 1990, Science 249:404-406; Cwirla et al., 1990, Proc. Natl. Acad. Sci. USA 87:6378-6382; and Felici, 1991, J. Mol. Biol. 222:301-310) (each of these references is incorporated herein in its entirety by reference). Antibodies or fragments thereof that have been identified to immunospecifically bind to a flaviviral antigen or a fragment thereof can then be assayed for their specificity and affinity for a flaviviral antigen.
The antibodies or fragments thereof may be assayed for immunospecific binding to a flaviviral antigen, particularly WNV antigen and cross-reactivity with other antigens by any method known in the art. Immunoassays which can be used to analyze immunospecific binding and cross-reactivity include, but are not limited to, competitive and non-competitive assay systems using techniques such as western blots, radioimmunoassays, ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, protein A immunoassays, to name but a few. Such assays are routine and well known in the art (see, e.g., Ausubel et al, eds, 2004, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York, which is incorporated by reference herein in its entirety). Exemplary immunoassays are described briefly below (but are not intended by way of limitation).
Immunoprecipitation protocols generally comprise lysing a population of cells in a lysis buffer such as RIPA buffer (1% NP-40 or Triton X-100, 1% sodium deoxycholate, 0.1% SDS, 0.15 M NaCl, 0.01 M sodium phosphate at pH 7.2, 1% Trasylol) supplemented with protein phosphatase and/or protease inhibitors (e.g., EDTA, PMSF, aprotinin, sodium vanadate), adding the antibody of interest to the cell lysate, incubating for a period of time (e.g., 1 to 4 hours) at 4° C., adding protein A and/or protein G sepharose beads to the cell lysate, incubating for about an hour or more at 4° C., washing the beads in lysis buffer and resuspending the beads in SDS/sample buffer. The ability of the antibody of interest to immunoprecipitate a particular antigen can be assessed by, e.g., western blot analysis. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the binding of the antibody to an antigen and decrease the background (e.g., pre-clearing the cell lysate with sepharose beads). For further discussion regarding immunoprecipitation protocols see, e.g., Ausubel et al, eds, 2004, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 10.16.1.
Western blot analysis generally comprises preparing protein samples, electrophoresis of the protein samples in a polyacrylamide gel (e.g., 8%-20% SDS-PAGE depending on the molecular weight of the antigen), transferring the protein sample from the polyacrylamide gel to a membrane such as nitrocellulose, PVDF or nylon, blocking the membrane in blocking solution (e.g., PBS with 3% BSA or non-fat milk), washing the membrane in washing buffer (e.g., PBS-Tween 20), blocking the membrane with primary antibody (the antibody of interest) diluted in blocking buffer, washing the membrane in washing buffer, blocking the membrane with a secondary antibody (which recognizes the primary antibody, e.g., an anti-human antibody) conjugated to an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) or radioactive molecule (e.g., 32P or 125I) diluted in blocking buffer, washing the membrane in wash buffer, and detecting the presence of the antigen. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected and to reduce the background noise. For fuirther discussion regarding western blot protocols see, e.g., Ausubel et al, eds, 2004, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 10.8.1.
ELISAs comprise preparing antigen, coating the well of a 96 well microtiter plate with the antigen, adding the antibody of interest conjugated to a detectable compound such as an enzymatic substrate (e.g., horseradish peroxidase or alkaline phosphatase) to the well and incubating for a period of time, and detecting the presence of the antigen. In ELISAs the antibody of interest does not have to be conjugated to a detectable compound; instead, a second antibody (which recognizes the antibody of interest) conjugated to a detectable compound may be added to the well. Further, instead of coating the well with the antigen, the antibody may be coated to the well. In this case, a second antibody conjugated to a detectable compound may be added following the addition of the antigen of interest to the coated well. One of skill in the art would be knowledgeable as to the parameters that can be modified to increase the signal detected as well as other variations of ELISAs known in the art. For further discussion regarding ELISAs see, e.g., Ausubel et al, eds, 2004, Current Protocols in Molecular Biology, Vol. 1, John Wiley & Sons, Inc., New York at 11.2.1.
The binding affinity of an antibody to an antigen and the off-rate of an antibody-antigen interaction can be determined by competitive binding assays. One example of a competitive binding assay is a radioimmunoassay comprising the incubation of labeled antigen (e.g., 3H or 125I) with the antibody of interest in the presence of increasing amounts of unlabeled antigen, and the detection of the antibody bound to the labeled antigen. The affinity of the antibody of the present invention or a fragment thereof for a WNV antigen and the binding off-rates can be determined from the data by scatchard plot analysis. Competition with a second antibody can also be determined using radioimmunoassays. In this case, a WNV antigen is incubated with an antibody of the present invention or a fragment thereof conjugated to a labeled compound (e.g., 3H or 125I) in the presence of increasing amounts of an unlabeled second antibody.
In a preferred embodiment, BIAcore kinetic analysis is used to determine the binding on and off rates of antibodies or fragments thereof to a WNV antigen. BIAcore kinetic analysis comprises analyzing the binding and dissociation of a WNV antigen from chips with immobilized antibodies or fragments thereof on their surface.
The antibodies or fragments thereof can also be assayed for their ability to inhibit the binding of a flaviviral antigen to its host cell receptor using techniques known to those of skill in the art and exemplified herein. For example, cells expressing the receptor for WNV can be contacted with WNV in the presence or absence of an antibody or fragment thereof and the ability of the antibody or fragment thereof to inhibit WNV's binding can measured by, for example, flow cytometry or a scintillation assay. WNV (e.g., WNV antigen such as E protein) or the antibody or antibody fragment can be labeled with a detectable compound such as a radioactive label (e.g., 32P, 35S, and 125I) or a fluorescent label (e.g., fluorescein isothiocyanate, rhodamine, phycoerythrin, phycocyanin, allophycocyanin, o-phthaldehyde and fluorescamine) to enable detection of an interaction between WNV and its host cell receptor. Alternatively, the ability of antibodies or fragments thereof to inhibit WNV from binding to its receptor can be determined in cell-free assays. For example, WNV or a WNV antigen can be contacted with an antibody or fragment thereof and the ability of the antibody or antibody fragment to inhibit WNV or the WNV antigen from binding to its host cell receptor can be determined. Preferably, the antibody or the antibody fragment is immobilized on a solid support and WNV or a WNV antigen is labeled with a detectable compound. Alternatively, WNV or a WNv antigen is immobilized on a solid support and the antibody or fragment thereof is labeled with a detectable compound. WNV or a WNV antigen may be partially or completely purified (e.g., partially or completely free of other polypeptides) or part of a cell lysate. Further, a WNV antigen may be a fusion protein comprising the WNV antigen and a domain such as glutathionine-S-transferase. Alternatively, a WNV antigen can be biotinylated using techniques well known to those of skill in the art (e.g., biotinylation kit, Pierce Chemicals; Rockford, Ill.).
The antibodies or fragments thereof are preferably tested in vitro, and then in vivo for the desired therapeutic or prophylactic activity, prior to use in humans. For example, in vitro assays which can be used to determine whether administration of a specific antibody or composition of the present invention is indicated, include in vitro cell culture assays in which a subject tissue sample is grown in culture, and exposed to or otherwise administered an antibody or composition of the present invention, and the effect of such an antibody or composition of the present invention upon the tissue sample is observed. In various specific embodiments, in vitro assays can be carried out with representative cells of cell types involved in a WNV infection to determine if an antibody or composition of the present invention has a desired effect upon such cell types. Preferably, the antibodies or compositions of the invention are also tested in in vitro assays and animal model systems prior to administration to humans. In a specific embodiment, mice are administered an antibody the invention or fragment thereof, or a composition of the invention, challenged with 100 to 1000 pfu of WNV, and four or more days later the mice are sacrificed and WNV titer and anti-WNV antibody serum titer is determined. In another specific embodiment, mice are administered 100 or 1000 pfu of WNV at day 0. At a point following infection, e.g., day 1, 2, 3, 4, 5, or 6, antibody or antibody variants are administered as a single does or as multiple doses. The therapeutic effect of the antibody or antibody variant can be assessed by monitoring, for example, the WNV titer, anti-WNV antibody titer, and/or the health and/or survival of the mice administered the antibody or antibody variant.
In accordance with the invention, clinical trials with human subjects need not be performed in order to demonstrate the prophylactic and/or therapeutic efficacy of antibodies or fragments thereof. In vitro and animal model studies using the antibodies or fragments thereof can be extrapolated to humans and are sufficient for demonstrating the prophylactic and/or therapeutic utility of said antibodies or antibody fragments.
Antibodies or compositions of the present invention for use in therapy can be tested for their toxicity in suitable animal model systems, including but not limited to rats, mice, cows, monkeys, and rabbits. For in vivo testing of an antibody or composition's toxicity any animal model system known in the art may be used.
Efficacy in treating or preventing viral infection may be demonstrated by detecting the ability of an antibody or composition of the invention to inhibit the replication of the virus, to inhibit transmission or prevent the virus from establishing itself in its host, to reduce the incidence of WNV infection, or to prevent, ameliorate or alleviate one or more symptoms associated with WNV infection. The treatment is considered therapeutic if there is, for example, a reduction is viral load, amelioration of one or more symptoms, a reduction in the duration of a WNV infection, or a decrease in mortality and/or morbidity following administration of an antibody or composition of the invention. Further, the treatment is considered therapeutic if there is an increase in the immune response following the administration of one or more antibodies or fragments thereof which immunospecifically bind to one or more WNV antigens.
Antibodies or compositions of the invention can be tested in vitro and in vivo for the ability to induce the expression of cytokines such as IFN-α, IFN-β, IFN-γ, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-12 and IL-15. Techniques known to those of skill in the art can be used to measure the level of expression of cytokines. For example, the level of expression of cytokines can be measured by analyzing the level of RNA of cytokines by, for example, RT-PCR and Northern blot analysis, and by analyzing the level of cytokines by, for example, immunoprecipitation followed by western blot analysis and ELISA.
Antibodies or compositions of the invention can be tested in vitro and in vivo for their ability to modulate the biological activity of immune cells, preferably human immune cells (e.g., T-cells, B-cells, and Natural Killer cells). The ability of an antibody or composition of the invention to modulate the biological activity of immune cells can be assessed by detecting the expression of antigens, detecting the proliferation of immune cells, detecting the activation of signaling molecules, detecting the effector function of immune cells, or detecting the differentiation of immune cells. Techniques known to those of skill in the art can be used for measuring these activities. For example, cellular proliferation can be assayed by 3H-thymidine incorporation assays and trypan blue cell counts. Antigen expression can be assayed, for example, by immunoassays including, but are not limited to, competitive and non-competitive assay systems using techniques such as western blots, immunohistochemistry radioimmunoassays, ELISA (enzyme linked immunosorbent assay), “sandwich” immunoassays, immunoprecipitation assays, precipitin reactions, gel diffusion precipitin reactions, immunodiffusion assays, agglutination assays, complement-fixation assays, immunoradiometric assays, fluorescent immunoassays, protein A immunoassays and FACS analysis. The activation of signaling molecules can be assayed, for example, by kinase assays and electrophoretic shift assays (EMSAs).
Antibodies or compositions of the invention can also be tested for their ability to inhibit viral replication or reduce viral load in in vitro, ex vivo and in vivo assays. Antibodies or compositions of the invention can also be tested for their ability to decrease the time course of WNV infection. Antibodies or compositions of the invention can also be tested for their ability to increase the survival period of humans suffering from WNV infection by at least 25%, preferably at least 50%, at least 60%, at least 75%, at least 85%, at least 95%, or at least 99%. Further, antibodies or compositions of the invention can be tested for their ability reduce the hospitalization period of humans suffering from WNV infection by at least 60%, preferably at least 75%, at least 85%, at least 95%, or at least 99%. Techniques known to those of skill in the art can be used to analyze the function of the antibodies or compositions of the invention in vivo.
Compounds for use in therapy can be tested in suitable animal model systems prior to testing in humans, including, but not limited to, in rats, mice, chicken, cows, monkeys, pigs, dogs, rabbits, hamsters, etc., for example, the animal models described above. Any animal system well-known in the art may be used.
Combinations of prophylactic and/or therapeutic agents can be tested in suitable animal model systems prior to use in humans. In a specific embodiment of the invention, combinations of prophylactic and/or therapeutic agents are tested in a mouse model system. Such model systems are widely used and well-known to the skilled artisan. Prophylactic and/or therapeutic agents can be administered repeatedly. Several aspects of the procedure may vary such as the temporal regime of administering the prophylactic and/or therapeutic agents, and whether such agents are administered separately or as an admixture.
Once the prophylactic and/or therapeutic agents of the invention have been tested in an animal model they can be tested in clinical trials to establish their efficacy. Establishing clinical trials will be done in accordance with common methodologies known to one skilled in the art, and the optimal dosages and routes of administration as well as toxicity profiles of the compositions of the invention can be established using routine experimentation.
The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosage of the prophylactic and/or therapeutic agents for use in humans. The dosage of such agents lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For any agent used in the method of the invention, the therapeutically effective dose can be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e., the concentration of the test compound that achieves a half-maximal inhibition of symptoms) as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.
Additional description of antibodies in general, as well as antibodies that bind WNV E protein, including the E16 monoclonal antibody, methods of making such antibodies, methods of using such antibodies, methods of characterizing such antibodies, and the like, may be found in U.S. Provisional Application No. 60/581,819 and U.S. application Ser. No. 11/159,046, filed Jun. 21, 2005, each of which is hereby incorporated by reference in its entirety for all purposes. In particular, all of the disclosure of these applications relating to antibodies, antibody derivatives, methods of making or characterizing, and all other disclosure relating to antibodies in general can be used to make, use, modify, manipulate, and otherwise engineer the antibodies of the invention as described herein.
5.11 Methods of Treating or Preventing WNV Infection
In yet another aspect, the invention provides a method of treating WNV infection. In certain embodiments, the methods comprising administering to a subject an effective amount of an antibody of the invention, as described herein. In certain embodiments, the antibody is a purified antibody. In certain embodiments, the antibody is not 5H10, 3A3, 5C5, 7H2, 11C, 17C8, 10C5, 8B10, E1, E16, E24, E27, E33, E34, E40, E43, E47, E49, and/or E58. In certain embodiments, the antibody is a monoclonal antibody.
In certain embodiments, the antibody binds the neutralizing epitope of domain III of WNV E protein with higher affinity than the antibody binds an enhancing epitope of domain III of WNV E protein. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 330-333 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering.
In certain embodiments, the antibody is administered to the subject in the form of a pharmaceutical composition.
In another aspect, the invention provides a method of preventing WNV infection, comprising administering to a subject an effective amount of an antibody of the invention. In certain embodiments, the antibody binds a neutralizing epitope of domain III of WNV E protein with higher affinity than the antibody binds an enhancing epitope of domain III of WNV E protein. In certain embodiments, the antibody is not 5H10, 3A3, 5C5, 7H2, 11C, 17C8, 10C5, 8B10, E1, E16, E24, E27, E33, E34, E40, E43, E47, E49, and/or E58. In certain embodiments, the antibody is a monoclonal antibody. In certain embodiments, the antibody binds the neutralizing epitope of domain III of WNV E protein with higher affinity than the antibody binds an enhancing epitope of domain III of WNV E protein. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 330-333 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the neutralizing epitope of the domain III of the WNV E protein comprises an amino acid sequence corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the antibody is administered to the subject in the form of a pharmaceutical composition.
5.12 WNV Immunogens
In yet another aspect, the invention provides WNV immunogens. The WNV immunogens of the invention are useful, for example, for inducing an immune response in a subject against a WNV epitope and, for example, in pharmaceutical compositions as described below. Preferably, the WNV epitope is a neutralizing epitope. The immune response that is induced preferably is effective to reduce, more preferably prevent, WNV infection in the subject following induction of the immune response.
Accordingly, in certain embodiments, the invention provides a WNV immunogen that comprises a peptide corresponding to amino acids 300-309, 330-333, 365-368, or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering, wherein the peptide is arranged in the three dimensional conformation of amino acids 300-309, 330-333, 365-368, or 389-391, respectively, of WNV E protein according to the structural coordinates shown in Table 7. In certain embodiments, the immunogen does not comprise the entire amino acid sequence of domain III of WNV E protein.
In certain embodiments, the WNV immunogen comprises a peptide corresponding to amino acids 300-309 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the WNV immunogen comprises a peptide corresponding to amino acids 330-333 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, immunogen comprises a peptide corresponding to amino acids 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises a peptide corresponding to amino acids 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 300-309 and 330-333 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 300-309 and 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 300-309 and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 330-333 and 365-368 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 330-333 and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 300-309, 330-333, and 365-368, of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 300-309, 330-333, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 300-309, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises peptides corresponding to amino acids 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In a preferred embodiment, the immunogen comprises peptides corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering. In certain embodiments, the immunogen comprises an amino acid sequence corresponding to amino acids 300-391 of WNV E protein, using SEQ ID NO:4 for numbering.
In certain embodiments, the peptide linkers comprise one or more glycines. In certain embodiments, the peptides corresponding to amino acids 300-309, 330-333, 365-368, and/or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering, are connected with each other with linkers. In general, the exact nature of the linker used in the WNV immunogens is unimportant, so long as the linker links the peptides of the WNV immunogen in a manner that permits the peptides to assume the proper molecular conformation, such as that presented in Table 7. In certain embodiments, the linker can form a covalent bond between the peptides of the WNV immunogen. In other embodiments, the linker can link peptides of the WNV immunogen with one or more non-covalent interactions of sufficient affinity. One of skill in the art can readily recognize linkers that interact with each other with sufficient affinity to be useful in the WNV immunogens of the invention. For example, biotin can be attached to a peptide of the WNV immunogen, and streptavidin can be attached to another peptide. In certain embodiments, the linker can directly link peptides of the WNV immunogen, e.g., via a covalent bond. In other embodiments, the linker itself comprises two or more molecules that associate in order to link the peptides of the WNV immunogen. Exemplary linkers include, but are not limited to, straight or branched-chain carbon linkers, heterocyclic carbon linkers, substituted carbon linkers, unsaturated carbon linkers, aromatic carbon linkers, peptide linkers, etc. In certain embodiments, the peptides corresponding to amino acids 300-309, 330-333, 365-368, and/or 389-391 of WNV E protein, using SEQ ID NO:4 for numbering, are connected with each other with peptide linkers. In certain embodiments, the peptide linkers comprise one or more amino acids selected from the group consisting of alanine (A), cysteine (C), aspartate (D), glutamate (E), glycine (G), isoleucine (I), lysine (K), leucine (L), methionine (M), asparagine (N), glutamine (Q), arginine (R), serine (S), threonine (T), and valine (V).
In embodiments where a linker is used to connect peptides of the WNV immunogen, the linkers can be attached to the peptides by any means or method known by one of skill in the art without limitation. For example, the linker can be attached to the peptides of the WNV immunogen with an ether, ester, thioether, thioester, amide, imide, disulfide or other suitable moieties. The skilled artisan can select the appropriate linker and means for attaching the linker based on the physical and chemical properties of the chosen linker. The linker can be attached to any suitable functional group on the molecule(s). For example, the linker can be attached to sulfhydryl (—S), carboxylic acid (COOH) or free amine (—NH2) groups, which are available for reaction with a suitable functional group on a linker. These groups can also be used to directly connect the peptides of the WNV immunogen in the absence of a linker.
Further, peptides of the WNV immunogen can be derivatized, for example, to facilitate attachment of a linker to these moieties. For example, such derivatization can be accomplished by attaching suitable derivatives such as those available from Pierce Chemical Company, Rockford, Ill. Alternatively, derivatization may involve chemical treatment of peptides of the WNV immunogen. For example, glycol cleavage of the sugar moiety of a carbohydrate attached to a WNV immunogen with periodate generates free aldehyde groups. These free aldehyde groups may be reacted with free amine or hydrazine groups on the remainder of the molecule in order to connect these portions of the molecule. See U.S. Pat. No. 4,671,958. Further, the skilled artisan can generate free sulfhydryl groups on proteins to provide a reactive moiety for making a disulfide, thioether, theioester, etc. linkage. See U.S. Pat. No. 4,659,839.
In certain embodiments, the peptides corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering, are expressed as a single polypeptide. In certain embodiments, the peptides corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering, are expressed as a plurality of peptides. In certain embodiments, the WNV immunogen comprises at least two cysteine residues that form a disulfide bond. In certain embodiments, the WNV immunogen comprises at least four cysteine residues that collectively form two disulfide bonds.
In certain embodiments, the WNV immunogen comprises an amino acid sequence corresponding to the variable domain of an immunoglobulin molecule, wherein the peptide corresponding to amino acids 330-333 of WNV E protein replaces one or more amino acids of the immunoglobulin molecule corresponding to CDR1; the peptide corresponding to amino acids 365-368 of WNV E protein replaces one or more amino acids of the immunoglobulin molecule corresponding to CDR2, and the peptide corresponding to amino acids 389-391 of WNV E protein replaces one or more amino acids of the immunoglobulin molecule corresponding to CDR3.
In certain embodiments, the WNV immunogen, when administered in to a subject, induces production of antibodies in the subject that bind domain III of WNV E protein. In certain embodiments, the WNV immunogen, when administered in to a subject, induces production of antibodies in the subject that neutralize infection by WNV.
The WNV immunogens can be prepared using any suitable method. In certain embodiments, the WNV immunogens can be prepared by chemical synthesis. In other embodiments, the WNV immunogens can be prepared biologically using suitable vectors in appropriate cell cultures as described below.
In certain embodiments, the WNV immunogens can be used in pharmaceutical compositions without further modification. In certain embodiments, the WNV immunogens can be modified, e.g., chemical conjugates, fusion proteins, pegylation, and the like. For example, chemical or nucleotidic or peptidic modifications can be made to allow the peptides to pass through certain biological barriers, to solubilize better, or to facilitate their incorporation into particular galenical forms, such as, e.g., liposomes or microparticles. Further, the WNV immunogens can be deglycosylated or glycosylated, as appropriate.
In certain embodiments, at least one portion of the WNV immunogens of the invention can be conjugated to a support onto which it is absorbed or bound in a covalent or non-covalent manner. The support can be, e.g., natural or synthetic carrier molecules. Such embodiments are useful, for example, in methods of purifying antibodies reactive with the antigens using WNV immunogens bountd to solid supports and in methods of inducing an immune response with the immunogens using immunogens bound to carrier molecules to increase the immunogenicity of the antigens. Preferably, the carrier molecules are physiologically acceptable and non toxic. The carrier molecules, in the context of an pharmaceutical composition, also preferably can increase the immunogenicity of the WNV immunogens. The WNV immunogens can be connected with the carrier molecules by, e.g., complementary reactive groups respectively present on the carrier molecule and the peptide known to those skilled in the art. Useful chemical compounds for coupling include, without limitation, dinitrophenol groups and arsonilic acid. Examples of carrier molecules include, but are not limited to, natural proteins such as, e.g., tetanus anatoxin, ovalbumin, serum albumin, hemocyamines, keyhole limpet hemocyanin (KLH), PPD (purified protein derivative) of tuberculin, red blood cells, tetanus toxoid, cholera toxoid, agarose beads, activated carbon, bentonite, etc.; synthetic macromolecular supports such as, e.g., polylysine or poly(D-L-alanine)-poly(L-lysine); hydrocarbon or lipid supports such as, e.g., saturated or unsaturated fatty acids. For a review of some general considerations in use of such compounds, see Harlow and Lane, eds., 1988, Antibodies, A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Further, liposomes, particles and microparticles, vesicles, latex bead microspheres, polyphosphoglycans (PGLA) or polystyrene can also be used as a support in the pharmaceutical compositions of the invention.
In yet another embodiment, the WNV immunogens may be in the form of a multiple antigenic peptide (“MAP”, also referred to as an octameric lysine core peptide) construct. Such a construct may be designed employing the MAP system described by Tam, 1988, Proc. Natl. Acad. Sci. USA 85:5409. In this system, multiple copies of a WNV antigen are synthesized onto a core matrix of lysine residues as described in Posnett et al., 1988, J. Biol. Chem. 263:1719; and Tam, 1992, “Chemically Defined Synthetic Immunogens and Vaccines by the Multiple Antigen Peptide Approach”, Vaccine Research and Developments, Vol. 1, Koff and Six, eds., Marcel Deblau, Inc., New York, N.Y., pp. 51-87. Each MAP contains multiple copies of one antigen.
Still other modified WNV immunogens may be prepared by any number of conventional techniques to improve production thereof, to enhance protein stability or other characteristics, e.g., binding activity or bioavailability, or to confer some other desired property upon the protein. Further, useful fragments of these polypeptides may be readily prepared by one of skill in the art using known techniques, such as deletion mutagenesis and expression.
The WNV immunogens of the present invention may also be constructed, using conventional genetic engineering techniques as part of a larger and/or multimeric protein or protein compositions. Antigens of this invention may be in combination with outer surface proteins or other proteins or antigens of other pathogens, such as those described herein, or various fragments of the antigens described herein may be in combination with each other. In such combination, the antigen may be in the form of a fusion protein. The antigen of the invention may be optionally fused to a selected polypeptide or protein derived from other microorganisms. For example, an antigen or polypeptide of this invention may be fused at its N-terminus or C-terminus to a polypeptide from another pathogen or to more than one polypeptide in sequence. Any polypeptides, without limitation, known by those of skill in the art to be useful for this purpose may be used in such embodiments.
A WNV immunogen of the present invention may also be used in the form of a pharmaceutically acceptable salt. Suitable acids and bases which are capable of forming salts with the polypeptides of the present invention are well known to those of skill in the art, and include inorganic and organic acids and bases.
5.12.1 Pharamaceutical Compositions Comprising a WNV Immunogen
In another aspect, the invention provides a pharmaceutical composition comprising a WNV immunogen of the invention and a pharmaceutically acceptable excipient, diluent, vehicle, or carrier. Any such pharmaceutically acceptable excipient, diluent, carrier, or vehicle known to one of skill in the art without limitation can be used. Examples of a suitable excipient, diluent, carrier, or vehicle can be found in Remington's Pharmaceutical Sciences, 19th Ed. 1995, Mack Publishing Co., Easton. Preferably, the pharmaceutical compositions induce a protective immune response when administered to a subject. In certain embodiments, the pharmaceutical composition is in a single unit dosage form. In certain embodiments, the pharmaceutical composition comprises a stabilizer. In certain embodiments, the pharmaceutical composition is formulated for storage for at least about 6 months without loss of more than about 20% activity. In certain embodiments, the pharmaceutical composition is an immunogenic composition.
The pharmaceutical compositions can be administered to a human for the treatment or prevention of WNV infection. Thus, the pharmaceutical compositions are generally compatible with administration to a human. In certain embodiments, the pharmaceutical compositions can be in any suitable solid or liquid form for pharmaceutical administration, e.g., in liquid administration forms, as a gel, or any other support allowing controlled release, for example. For example, the pharmaceutical composition can be an injectable composition, e.g., a pharmaceutical composition formulated for injection into the blood in a human.
The pharmaceutical compositions of the invention can also comprise components that increase the immunogenicity of a WNV immunogen. For example, the pharmaceutical compositions can comprise immunogenic peptides other than WNV immunogens, specific or nonspecific immunogenic adjuvants such as, e.g., alum, aluminum hydroxide, aluminum phosphate, QS21, Freund's adjuvant, SBA2 adjuvant, montanide, polysaccharides, lipopolysaccharides, lipopolysaccharide derivatives, lipid A, CpG-containing nucleic acids, non-CpG containing nucleic acids, oil-in-water emulsions, or equivalent compounds. Other suitable adjuvants are described in Sheikh et al., 2000, Cur. Opin. Mol. Ther. 2:37-54. The immunogen may also be modified by other techniques, such as denaturation with heat and/or SDS.
The pharmaceutical compositions of the invention may comprise one or a plurality of WNV immunogens. For example, a pharmaceutical composition can include WNV immunogens from several circulating strains of a pathogen, e.g., from more than one WNV strain or from more than one flavivirus. In certain embodiments, the pharmaceutical composition can comprise a combination of WNV immunogens from two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, or more strains of WNV. In certain embodiments, the pharmaceutical composition can comprise a combination of immunogens from two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, or more different flaviviruses. In certain embodiments, the pharmaceutical composition comprises a combination of WNV immunogens that, when administered to a subject, induces an immune response in the subject that recognizes an E protein from more than one WNV strain. In certain embodiments, the pharmaceutical composition comprises a combination of immunogens that, when administered to a subject, induces an immune response in the subject that recognizes an E protein from more than one flavivirus.
The pharmaceutical compositions are generally formulated appropriately for the immediate use intended for the composition. For example, if the composition is not to be administered immediately, it can be formulated suitably for storage. One such composition is a lyophilized preparation of the WNV immunogen(s) together with a suitable stabilizer. Alternatively, the pharmaceutical composition can be formulated for storage in a solution with one or more suitable stabilizers. Any such stabilizer known to one of skill in the art without limitation can be used. For example, stabilizers suitable for lyophilized preparations include, but are not limited to, sugars, salts, surfactants, proteins, chaotropic agents, lipids, and amino acids. Stabilizers suitable for liquid preparations include, but are not limited to, sugars, salts, surfactants, proteins, chaotropic agents, lipids, and amino acids. Specific stabilizers than can be used in the pharmaceutical compositions include, but are not limited to, trehalose, serum albumin, phosphatidylcholine, lecithin, and arginine. Other compounds, compositions, and methods for stabilizing a lyophilized or liquid preparation of the WNV immunogens may be found, for example, in U.S. Pat. Nos. 6,573,237; 6,525,102; 6,391,296; 6,255,284; 6,133,229; 6,007,791; 5,997,856; and 5,917,021.
In an alternate embodiment, the pharmaceutical compositions may also comprise nucleic acids that express one or more WNV immunogen(s) described herein. For example, when injecting naked DNA encoding a WNV immunogen as described herein, this injection can result in expression of the encoded immunogen and an immune response against the immunogen. It is also possible to use naked DNA systems that comprise expression system or expression vectors. The expression vectors can in some cases improve the activity of the expressed immunogens. Any suitable immunization system employing DNA known by one of skill in the art, whether as part of an expression system or not, can be used to administer DNA to a subject. Examples of such immunization systems can be found, for example, in International Patent Publication No. WO 95/111307 and in Bot et al., 1996, Viral Immunol 9:207. Additional exemplary vectors for in vivo gene delivery and include, e.g., adeno-associated virus (International patent application No. PCT/US91/03440), adenovirus vectors (Kay et al., 1994, Proc. Natl. Acad. Sci. USA 91:2353; Ishibashi et al., 1993, J. Clin. Invest. 92:883), or other viral vectors, e.g., various poxviruses, vaccinia, etc. Methods for insertion of a desired gene, e.g., a WNV immunogen and obtaining in vivo expression of the encoded protein are well known to those of skill in the art. In certain embodiments, vectors incorporating sequences that are capable of increasing the immunogenicity of the WNV immunogens of the present invention, such as CpG sequences, the GMCSF (granulocyte macrophage colony stimulating factor) gene, or cytokine genes can be employed. The specific constructions depend on the host, the epitope and on the vector selected.
5.12.2 Kits Comprising Pharmaceutical Compositions
In yet another aspect, the invention provides a kit comprising a pharmaceutical composition of the invention. In certain embodiments, the kit comprises a pharmaceutical composition is in a single unit dosage form. In certain embodiments, the kit further comprises instructions directing administration of the pharmaceutical composition to a subject. In certain embodiments, the kit further comprises instructions directing a medical professional to administer the pharmaceutical composition to a subject.
5.12.3 Dosage and Regimen of WNV Immunogens
Generally, a pharmaceutically effective amount of the WNV immunogens of the invention is administered to a subject. The skilled artisan can readily determine if the dosage of the WNV immunogen is sufficient to elicit an immune response by monitoring the immune response so elicited, as described below. In certain embodiments, an amount of pharmaceutical composition corresponding to between about 1 μg and about 1000 μg of WNV immunogen is administered. In other embodiments, an amount of pharmaceutical composition corresponding to between about 10 μg and about 500 μg of WNV immunogen is administered. In still other embodiments, an amount of pharmaceutical composition corresponding to between about 10 μg and about 250 μg of WNV immunogen is administered. In yet other embodiments, an amount of pharmaceutical composition corresponding to between about 10 μg and about 100 μg of WNV immunogen is administered. In certain embodiments, an amount of pharmaceutical composition corresponding to between about 10 μg and about 50 μg of WNV immunogen is administered. Further guidance on selecting an effective dose of the pharmaceutical compositions may be found, for example, in Rose and Friedman, 1980, Manual of Clinical Immunology, American Society for Microbiology, Washington, D.C.
The volume of pharmaceutical composition administered will generally depend on the concentration of WNV immunogen and the formulation of the composition. In certain embodiments, a unit dose of the pharmaceutical composition is between about 0.05 ml and about 1 ml, preferably about 0.5 ml. The pharmaceutical compositions can be prepared in dosage forms containing between 1 and 50 doses (e.g., 0.5 ml to 25 ml), more usually between 1 and 10 doses (e.g., 0.5 ml to 5 ml)
The pharmaceutical compositions of the invention can be administered in one dose or in multiple doses. A dose can be followed by one or more doses spaced by about 4 to about 8 weeks, by about 1 to about 3 months, or by about 1 to about 6 months. Additional booster doses can be administered as needed. In certain embodiments, booster doses are administered in about 1 to about 10 years.
5.12.4 Administration of WNV Immunogens
The WNV immunogens of the invention can be administered to a subject by any method known to one of skill in the art. In certain embodiments, the pharmaceutical compositions are injected into the subject. The pharmaceutical composition can be injected subcutaneously, intramuscularly, intravenously, intra-arterially, or parenterally. In such embodiments, the pharmaceutical composition preferably comprises an adjuvant, as described above.
5.12.5 Methods of Making WNV Immunogens
Any suitable expression system known by one of skill in the art for producing a peptide, polypeptide, or nucleic acid immunogen can be used to produce the WNV immunogens of the invention. Alternately, the WNV immunogens can be chemically synthesized, either in whole or in part. To produce recombinant WNV immunogens, the nucleic acid sequences encoding the immunogens can be inserted into a suitable expression system. Desirably, a recombinant molecule or vector can be constructed in which the polynucleotide sequence encoding the selected protein, e.g., a WNV immunogen, is operably linked to a heterologous expression control sequence permitting expression of the protein. Numerous types of appropriate expression vectors are known in the art for protein expression by standard molecular biology techniques. Such vectors can be selected from among conventional vector types including insects, e.g., baculovirus expression, or yeast, fungal, bacterial or viral expression systems. Other appropriate expression vectors, of which numerous types are known in the art, can also be used for this purpose. Methods for obtaining such expression vectors are well-known. See, e.g., Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., and Ausubel et al., eds., Current Edition, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, NY, and references cited therein.
Suitable host cells or cell lines for transfection by this method include bacterial cells, such as, for example, E. coli (e.g., HB101, MC 1061, etc.) B. subtilis, Pseudomonas ssp., Streptomyces ssp., and the like; and mammalian cells, such as, for example, human 293 cells, Chinese hamster ovary cells (CHO), monkey COS-1 cells, and murine 3T3 cells. Indeed, any suitable host cell, method for transfection, culture, amplification, screening, production, purification, etc. known to one of skill in the art without limitation can be used to produce a WNV immunogen. Further, strains of yeast cells or other ftngal systems known to those skilled in the art are also available as host cells for expression of the WNV immunogens of the present invention. Alternatively, insect cells such as Spodoptera frugipedera (Sf9) cells may be used.
Thus, the present invention provides a method for producing a WNV immunogen that comprises transfecting, e.g., by conventional means such as electroporation, a host cell with at least one expression vector containing a polynucleotide encoding the WNV immunogen under the control of a transcriptional regulatory sequence. The transfected or transformed host cell can be then cultured under conditions that allow expression of the protein. The expressed protein can be recovered, isolated, and optionally purified from the cell (or from the culture medium, if expressed extracellularly) by appropriate means known to one of skill in the art. For example, an antibody that specifically binds a WNV immunogen, e.g., E16, can be conveniently used to affinity purify a WNV inmmunogens of the invention.
For example, the immunogens can be isolated in soluble form following cell lysis, or extracted using known techniques, e.g., in guanidine chloride. If desired, the proteins or fragments of the invention are produced as a fusion protein such as those described above. Alternatively, for example, it may be desirable to produce fusion proteins to enhance expression of the immunogen in a selected host cell, to improve purification, or for use in monitoring the presence of the immunogen in tissues, cells or cell extracts. Suitable fusion partners for the proteins of the invention are well known to those of skill in the art and include, among others, β-galactosidase, glutathione-S-transferase, poly-histidine and maltose binding protein.
Thus, the invention also provides a method for preparing a WNV immunogen of the invention by transforming a host cell using an expression vector (plasmid, cosmid, virus, etc.) comprising DNA sequences encoding the immunogens of the invention, and culturing the transformed host cell and recovering the peptide in the culture medium.
The invention further provides a vector (either cloning and/or expression) and a host cell (prokaryotic or eukaryotic) transformed by the vector and comprising regulating elements allowing expression of the nucleotide sequence coding for a immunogen of the invention.
5.12.6 Polynucleotides Encoding WNV Immunogens
In another aspect, the invention provides polynucleotides comprising a nucleotide sequence encoding a WNV immunogen of the invention as described herein. These polynucleotides are useful, for example, for making the WNV immunogens. In certain embodiments, the recombinant polynucleotides are based on polynucleotides encoding a WNV immunogen, as described herein. In other embodiments, the recombinant polynucleotides are based on polynucleotides that hybridize to a polynucleotide that encodes a WNV immunogen under stringent hybridization conditions.
In vitro methods that can be used to prepare a polynucleotide encoding WNV immunogens of the invention include, but are not limited to, reverse transcription, the polymerase chain reaction (PCR), the ligase chain reaction (LCR), the transcription-based amplification system (TAS), the self-sustained sequence replication system (3SR) and the QP replicase amplification system (QB). Any such technique known by one of skill in the art to be useful in construction of recombinant nucleic acids can be used. For example, a polynucleotide encoding the protein or a portion thereof can be isolated by polymerase chain reaction of cDNA using primers based on the DNA sequence of PE or another polynucleotide encoding a receptor binding domain.
Guidance for using these cloning and in vitro amplification methodologies are described in, for example, U.S. Pat. No. 4,683,195; Mullis et al., 1987, Cold Spring Harbor Symp. Quant. Biol. 51:263; and Erlich, ed., 1989, PCR Technology, Stockton Press, NY. Polynucleotides encoding a WNV immunogen or a portion thereof also can be isolated by screening genomic or cDNA libraries with probes selected from the sequences of the desired polynucleotide under stringent, moderately stringent, or highly stringent hybridization conditions.
Further, the polynucleotides can also encode a secretory sequence at the amino terminus of the encoded WNV immunogen. Such constructs are useful for producing the WNV immunogens in mammalian cells as they simplify isolation of the immunogen.
Furthermore, the polynucleotides of the invention also encompass derivative versions of polynucleotides encoding a WNV immunogen. Such derivatives can be made by any method known by one of skill in the art without limitation. For example, derivatives can be made by site-specific mutagenesis, including substitution, insertion, or deletion of one, two, three, five, ten or more nucleotides, of polynucleotides encoding the WNV immunogen. Alternatively, derivatives can be made by random mutagenesis. One method for randomly mutagenizing a nucleic acid comprises amplifying the nucleic acid in a PCR reaction in the presence of 0.1 mM MnCl2 and unbalanced nucleotide concentrations. These conditions increase the misincorporation rate of the polymerase used in the PCR reaction and result in random mutagenesis of the amplified nucleic acid.
5.12.7 Expression Vectors
In still another aspect, the invention provides expression vectors for expressing the WNV immunogens. Generally, expression vectors are recombinant polynucleotide molecules comprising expression control sequences operatively linked to a nucleotide sequence encoding a polypeptide. Expression vectors can readily be adapted for function in prokaryotes or eukaryotes by inclusion of appropriate promoters, replication sequences, selectable markers, etc. to result in stable transcription and translation of mRNA. Techniques for construction of expression vectors and expression of genes in cells comprising the expression vectors are well known in the art. See, e.g., Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., and Ausubel et al., eds., Current Edition, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, NY.
Useful promoters for use in expression vectors include, but are not limited to, a metallothionein promoter, a constitutive adenovirus major late promoter, a dexamethasone-inducible MMTV promoter, a SV40 promoter, a MRP pol III promoter, a constitutive MPSV promoter, a tetracycline-inducible CMV promoter (such as the human immediate-early CMV promoter), and a constitutive CMV promoter.
The expression vectors should contain expression and replication signals compatible with the cell in which the WNV immunogens are expressed. Expression vectors useful for expressing WNV immunogens include viral vectors such as retroviruses, adenoviruses and adenoassociated viruses, plasmid vectors, cosmids, and the like. Viral and plasmid vectors are preferred for transfecting the expression vectors into mammalian cells. For example, the expression vector pcDNA1 (Invitrogen, San Diego, Calif.), in which the expression control sequence comprises the CMV promoter, provides good rates of transfection and expression into such cells.
The expression vectors can be introduced into the cell for expression of the WNV immunogens by any method known to one of skill in the art without limitation. Such methods include, but are not limited to, e.g., direct uptake of the molecule by a cell from solution; facilitated uptake through lipofection using, e.g., liposomes or immunoliposomes; particle-mediated transfection; etc. See, e.g., U.S. Pat. No. 5,272,065; Goeddel et al., eds, 1990, Methods in Enzymology, vol. 185, Academic Press, Inc., CA; Krieger, 1990, Gene Transfer and Expression: A Laboratory Manual, Stockton Press, NY; Sambrook et al., 2001, Molecular Cloning: A Laboratory Manual, 3rd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y., and Ausubel et al., eds., Current Edition, Current Protocols in Molecular Biology, Greene Publishing Associates and Wiley Interscience, NY.
The expression vectors can also contain a purification moiety that simplifies isolation of the protein. For example, a polyhistidine moiety of, e.g., six histidine residues, can be incorporated at the amino terminal end of the protein. The polyhistidine moiety allows convenient isolation of the protein in a single step by nickel-chelate chromatography. In certain embodiments, the purification moiety can be cleaved from the remainder of the WNV immunogen following purification. In other embodiments, the moiety does not interfere with the function of the functional domains of the WNV immunogen and thus need not be cleaved.
5.12.8 Cells for Expressing a WNV Immunogen
In yet another aspect, the invention provides a cell comprising an expression vector for expression of the WNV immunogens, or portions thereof. The cell is preferably selected for its ability to express high concentrations of the WNV immunogen to facilitate purification of the protein. In certain embodiments, the cell is a prokaryotic cell, for example, E. coli. 
In other embodiments, the cell is a eukaryotic cell. Useful eukaryotic cells include yeast and mammalian cells. Any mammalian cell known by one of skill in the art to be useful for expressing a recombinant polypeptide, without limitation, can be used to express the WNV immunogens. For example, Chinese hamster ovary (CHO) cells can be used to express the WNV immunogens.
5.13 Methods of Inducing an Immune Response Against WNV DIII
In another aspect, the invention provides methods of inducing an immune response in a subject against domain III of WNV E protein. In general, the methods comprise administering a WNV immunogen of the invention to a subject in whom the immune response is to be induced. Typically, such WNV immunogens are administered in the form of a pharmaceutical composition, as described herein. In certain embodiments, the immune response that is induced is a prophylactic immune response, i.e., the subject is not already afflicted with WNV infection. In other embodiments, the immune response that is induced is therapeutic, i.e., the subject is already afflicted with WNV infection. The resultant immune responses can protect against infection by WNV or against cells that express WNV immunogens. For example, the immune response can be mounted against WNV, itself, or against cells containing intracellular WNV.
In certain embodiments, the methods comprise administering to a subject a WNV immunogen that comprises one or more peptides corresponding to amino acids 300-309, 330-333, 365-368, and 389-391 of WNV E protein, using SEQ ID NO:4 for numbering, wherein the peptide(s) is (are) arranged in the three dimensional conformation of amino acids 300-309, 330-333, 365-368, and 389-391, respectively, of WNV E protein according to the structural coordinates shown in Table 7. In certain embodiments, the immunogen does not comprise the entire amino acid sequence of domain III of WNV E protein.
In certain embodiments, the immune response induced in the subject is effective to prevent a WNV infection in the subject following exposure to WNV. In certain embodiments, the immune response that is induced is effective to prevent WNV from fusing with a cell that expresses a receptor ordinarily bound by a WNV. In certain embodiments, the immune response that is induced is effective to prevent infection of the cell expressing a receptor ordinarily bound by WNV but is not effective to prevent attachment of the WNV to the cell. In certain embodiments, an antibody produced by the immune response in the subject neutralizes WNV infection. In certain embodiments, the induced immune response is effective to reduce the severity of a WNV infection in the subject. In certain embodiments, the induced immune response is effective to treat a WNV infection in the subject. In certain embodiments, the induced immune response is effective to treat a disease, or a symptom thereof, mediated by WNV infection in the subject. In certain embodiments, the induced immune response is effective to treat or prevent one or more symptoms associated with WNV infection in a subject.
In certain embodiments, the subject is a mammal or bird. In certain embodiments, the subject is a goat, cow, rabbit, mouse, rat, horse, pig, ferret, weasel, or primate. In certain embodiments, the subject is a human.
In certain embodiments, the WNV immunogen is administered in a single dose. In certain embodiments, the WNV immunogen is administered in multiple doses. In certain embodiments, a WNV immunogen can be administered at a first time, then a WNV immunogen can be administered at a later second time. In certain embodiments, the administrations can be separated by one, two, three, four, five, six, nine, twelve, fifteen, or eighteen months or one, two, three, four, five, six, seven, eight, nine, ten, or more years. In certain embodiments, the subject's immune response against a WNV immunogen or a WNV can be assessed to determine whether the subject would benefit from a repeated (e.g., booster) administration of a WNV immunogen. In certain embodiments, the WNV immunogen administered at the first time is the same WNV immunogen as the immunogen administered at the later second time. In certain embodiments, the WNV immunogen administered at the first time is a different WNV immunogen from the immunogen administered at the later second time.
In certain embodiments, the methods comprise administering a WNV immunogen of the invention to a subject two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, fifteen, twenty, or more times. In certain embodiments, the WNV immunogen can be administered at regular intervals until death of the subject. The administration can be by any method described herein. Further, the administrations can be separated by as little as, e.g., about 1 month or as much as e.g., about ten or more years. In certain embodiments, the administrations can be at regular periods, e.g., about every six months, about every year, about every 18 months, about every two years. In other embodiments, the administrations can be irregular, e.g., a second administration after about three months of the first administration, then a third administration at about 2 years after the first. In still other embodiments, the administration can be variously irregular and regular, e.g., a second administration after about three months of the first administration, then a third administration at about 2 years after the first and further administrations every two years thereafter. In certain embodiments, the administrations are separated by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 months. In certain embodiments, the administrations are separated by about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 years.
6. EXAMPLES
The following examples are provided to illustrate aspects of the invention, and are not intended to limit the scope of the invention in any way.
6.1 Example 1
Preparation of Crystals of WNV DIII-E16 Fab Complex
The subsections below describe the production of WNV DIII polypeptide; E16 Fab fragments, and the preparation and characterization of diffraction quality WNV DIII-E16 Fab complex crystals.
6.1.1 Production and Purification of WNV DIII
This example describes the expression and purification of a polypeptide corresponding to domain III of WNV E protein. A plasmid for expressing WNV DIII, pWNV-DIII, was constructed in the following manner: PCR primers that correspond to nucleotides 1852-1866 as the forward primer and nucleotides 2201-2211 as the reverse primer with added 5′ BamH I and 3′ Xho I sites were used to PCR a 359 nucleotide fragment from an infectious cDNA clone of WNV generated from the NY1999 strain. After restriction digest, domain III of WNV was cloned into a modified PET21 vector (Novagen) that contains histidine and BirA tags at the amino terminus. After sequencing, a clone with complete identity to the parent domain III sequence was transformed into BL21 E. coli cells.
BL21 bacteria that contained the pWNV-DIII plasmid were grown overnight at 37° C. as a six-liter batch. After centrifugation (7,500×g for 30 minutes) and lysis of cells by sonication, inclusion bodies were isolated. Refolding of WNV-DIII was performed according to the protocol described in Carayannopoulos et al., 2002, J. Immunol. 169:4079-83. BL21 cells containing the WNV-DIII plasmid were grown to an optical density of 0.8 at 595 nm in Luria broth medium at 37° C. Bacteria were then induced with 0.5 mM isopropyl thiogalactoside (IPTG). After 4 hours, the cell pellets were harvested and suspended in 50 mM Tris-HCl, pH 8.0, 0.2 M NaCl, 5 mM EDTA and 5 mM DTT. Bacteria were lysed after lysozyme addition (final concentration of 1 mg/ml), and the viscosity was reduced by sonication. The WNV-DIII protein was obtained as insoluble aggregates in inclusion bodies, which were washed once in 50 mM Tris-HCl, pH 8.0, 0.1 M NaCl, 1 mM EDTA, 1 mM DTT, and 0.5% (v/v) Triton X-100, and once in 50 mM Tris-HCl, pH 8.0, 2 M NaCl, 1 M urea, 1 mM EDTA and 1 mM DTT. 0.5 mls of an 80% slurry of inclusion body was solubilized in 5 ml of 20 mM Tris (pH 8.0), 6M Guanidine HCl and 1 mM DTT; insoluble material was removed by centrifugation.
The remaining soluble WNV-DIII was refolded by slowly diluting out the denaturing reagents according to the protocol of Miley et al., 2003, J. Immunol. 170:6090-8. Briefly, 1.6 ml of guanidine HCl-solubilized WNV-DIII was added dropwise to 0.5 liter of 0.1 M Tris-HCl, pH 8.5, 400 mM L-arginine, 2 mM EDTA, 5 mM reduced glutathione, 0.5 mM oxidized glutathione, 0.1 mM phenylmethylsulfonyl fluoride. Additional aliquots were added every 4 h. After 24 h, the refolding mixture was concentrated under nitrogen to a volume of 2 ml. Refolded WNV-DIII was separated from aggregates on a Superdex 75 16/60 size exclusion column (Amersham Biosciences), treated with thrombin to remove the N-terminal BirA biotinylation tag and re-purified by size excusion chromatography. WNV-DIII was concentrated using a centricon-10 spin column and buffer exchanged into 10 mM Hepes pH 7.3, 100 mM NaCl, and 0.02% NaN3 
6.1.2 Production and Purification of E16 Fab Fragments
Fab fragments of E16 (IgG2b) were generated from purified IgG after papain digestion, protein A affinity chromatography and gel filtration chromatography according to the protocol presented in Harlow and Lane, 1988, Antibodies, A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor. E16 is expressed by a hybridoma assigned ATCC Accession No. PTA-6050 and can be obtained from this hybridoma according to standard techniques. The purity of Fab fragments was confirmed by non-reducing SDS-PAGE and silver staining.
6.1.3 Preparation of WNV DIII-E16 Fab Complex Crystals
Fab fragments of E16 were mixed with WNV-DIII in an equimolar ratio for 2 hours at 4° C., and then isolated after gel filtration (Superdex 75) chromatography. Fab-WNV-DIII complexes were concentrated using a centricon-10 spin column to 10-15 mg/ml and used for crystallization experiments.
The E16 Fab-DIII complex was crystallized by hanging drop vapor diffusion at 20° C. from a 1.5 μl drop containing a mixture of 0.5 μl protein in 20 mM HEPES (pH 7.5), 0.01% NaN3, 0.5 μl of 0.1 M HEPES (pH 8.5), 19% PEG 4000, 0.1M glycine and 0.5 μl of deionized water over wells containing precipitant solution. The E16-DIII crystals were dragged through a drop that contained well solution and 20% ethylene glycol and was rapidly cooled in a gaseous stream of liquid nitrogen at 100K. The crystals were annealed by blocking the cryostream twice for five seconds each.
6.2 Analysis and Characterization of WNV DIII-E16 Fab Complex Crystals
This example describes the methods used to generate and collect diffraction data from WNV DIII-E16 Fab complex crystals and determine the structure of the WNV DIII-E16 Fab complex from such data.
6.2.1 Diffraction Data Collection
A dataset was collected at ALS beamline 4.2.2 (Lawrence Berkeley Laboratories, Berkeley, Calif.) by the oscillation method at a wavelength of 1.5498 Å with a large-area CCD X-ray detector. Data was processed, scaled and merged with d*trek. See Pflugrath, 1999, Acta Crystallogr. D. Biol. Crystallogr. 55:1718-25. The crystals were in the space group P212121 with unit cell dimensions of a=52.4 Å, b=83.3 Å and c=110.6 Å with one molecule of WNV-DIII, one light chain of E16 and one papain cleaved heavy chain of E16 in the asymmetric unit.
6.2.2 Structure Determination and Analyses
The structure of WNV DIII-E16 Fab complex was determined using molecular replacement in molrep of CCP4i (see Vagin & Teplyakov, 2000, Acta Crystallogr D Biol Crystallogr. 56:1622-4) using the variable domain of an IgG1 Fab′ (PDB ID 2IGF) as a model for the variable domains, then the constant domains of and IgG1 Fab′ (PDB ID 2IGF) as a model for the constant domains, and finally the averaged domain III of WNV (PDB ID 1S6N) for the DIII fragment. The initial Rcryst and correlation coefficient were 43.3 and 56.3, respectively. The models were iteratively altered to the correct sequence using the program InsightII (Accelrys). The models were built in O (see Jones et al., 1991, Acta Crystallogr. 47:753-70) and refined in CNS (see Brunger et al., 1998, Acta Crystallogr. D. Biol. Crystallogr. 54:905-21) against all F>0. The final model contained 788 amino acids (residues 1-212 of chain L, 1-228 of chain H and residues 300-400 of chain A), 255 water molecules and one sodium ion. The final Rcryst and Rfree were 20.5 and 28.1, respectively after restrained B factor and positional refinement. A summary of these data is shown in Table 5. The refinement statistics are shown in Table 6; the accuracy of the model is reflected by the Ramachandran plot (FIG. 8) which disallowed only a single amino acid residue (Ala 51L), belonging to the L2 hypervariable loop of the VL.







TABLE 5





Summary of Data Collection


Data Collection for WNV DIII-E16 complexa


















Space Group
P2(1)2(1)2(1)



Unit Cell (Å3)
a = 52.4 b = 83.3 c = 110.6







Data Set
Native







Wavelength(Å)
1.5498



X-ray Source
ALS



Resolution(Å) (outer shell)
50–2.50 (2.59–2.50)*



Observations/Unique
59923/16985d



Completeness (%)
97.6 (82.7)c



Redundancy
3.53 (1.99)



Rsymb (%)
 8.3 (30.6)c



I/σ
11.3 (2.7)c







aValues as defined in d * trek (Pflugrath, JW)



bRsym = 100 × ΣhΣi|Ii(h) − <I(h)>|/ΣhΣiIi(h)



cValue in parentheses is for the highest resolution shell.



dI(+h) and I(−h) processed as independent reflections. Anomalous scattering contributions were included.













TABLE 6





Refinement Statistics


















Reflections Rworka/Rfree
16752/817 



#Protein Atomsb/Solvent
4044/255 



Rwork overall(outer shell) (%)
20.8 (25.9)c



Rfree overall(outer shell) (%)
28.2 (36.7)c



Rmsd Bond lengths (Å)/angles(°)
0.007/1.5 



Rmsd Dihedral/Improper (°)
27.3/0.83



Ramachandran plot



Most Favored/Additional (%)
87.1/12.2



Generous/Disallowed (%)
0.4/0.2



No. Protein Atoms
4040



No. Water Atoms
256



Average B-values (protein)
32.7



Average B-values (water)
32.4



Est. Coordinate Error (Å)
0.44







aR-value = 100 × Σh||Fobs(h)| − |Fcalc(h)||/Σh|Fobs(h)| for reflections with Fobs > 2σ.



bFor bonded protein atoms.



cValue in parentheses is the free R-value (Brunger, 1992, “Free R value: a novel statistical quantity for assessing the accuracy of crystal structures,” Nature 355: 472–475) determined from 5% of the data






Table 7, following below, provides the atomic structure coordinates of WNV DIII-E16 Fab complex. In the Table, coordinates for one WNV DIII polypeptide and one E16 Fab fragment, comprising one E16 light chain and one papain cleaved heavy chain of E16, comprising the asymmetric unit are provided. The amino acid residue numbers coincide with those used in FIGS. 1-3. In the first molecule of the WNV DIII-E16 Fab complex, the residue number is preceded by a 1, i.e., residue number 464 of the WNV DIII molecule of the dimer is denoted by “1464;” residue number 464 of the E16 light chain is denoted by “2464;” and residue number 464 of the E16 papain-cleaved heavy chain is denoted by “3464.”
The following abbreviations are used in Table 7:
“Atom Type” refers to the element whose coordinates are provided. The first letter in the column defines the element.
“A.A.” refers to amino acid.
“X, Y and Z” provide the Cartesian coordinates of the element.
“B” is a thermal factor that measures movement of the atom around its atomic center.
“OCC” refers to occupancy, and represents the percentage of time the atom type occupies the particular coordinate. OCC values range from 0 to 1, with 1 being 100%.
“PRT1” or “PRT2” relate to occupancy, with PRTI designating the coordinates of the atom when in the first conformation and PRT2 designating the coordinates of the atom when in the second or alternate conformation.
Structures coordinates for WNV DIII-E16 Fab complex according to Table 7 may be modified by mathematical manipulation. Such manipulations include, but are not limited to, crystallographic permutations of the raw structure coordinates, fractionalization of the raw structure coordinates, integer additions or subtractions to sets of the raw structure coordinates, inversion of the raw structure coordinates and any combination of the above.
6.2.3 Summary of Conclusions Regarding the Structure of WNV DIII-E16 Fab Complex
The structure determined above revealed that WNV DIII adopted an Ig-like β-sandwich topology similar to that found in other flavivirus E proteins, while the E16 Fab adopts a typical quaternary assembly (FIG. 9). The binding interface had a high degree of shape complementarily (Sc=0.763)9 and occludes 1550 A2 of surface area, with VH accounting for 67% of the total antibody-combining site (FIG. 10). E16 contacted DIII with 18 residues spread along all 6 of its CDR loops in addition to three VH framework residues (Tables 1, 2, and 3). The interaction between E16 and DIII was dominated by hydrogen bonds, with 16 direct hydrogen bonds and numerous water mediated networks at the interface of the complex.
E16 engaged four discontinuous segments of DIII, including the N-terminal region (residues E302-E309) and three strand-connecting loops: BC (E330-E333), DE (E365-E368) and FG (E389-E391). E16 contacted a total of 16 DIII residues, which together formed a single convex surface patch. Strikingly, yeast surface display epitope mapping of DIII identified four residues at the core of this binding site that are critical for E16 recognition (FIG. 12). To perform these yeast display mapping studies, an error-prone PCR derived library of WNV E (residues 296-415) was on the surface of yeast as Aga2 fusion proteins. See Chao et al., 2004, J Mol Biol 342:539-50 and Oliphant et al., 2005, Nat Med 11:522-30 (2005). Yeasts were screened for selective loss of E16 binding relative to other DIII specific mAbs by multiple rounds of fluorescence-activated cell sorting. Isolated clones were recovered, sequenced and evaluated for binding to E16 and the non-neutralizing, WNV DIII-specific mAb E22.
Non-conservative substitution at SerE306, LysE307, ThrE330 or ThrE332 disrupted E16 binding but not that of a non-neutralizing DIII-specific mAb, E22. These four residues clustered at the center of the E16IDIII interface (FIG. 10). The decreased binding associated with mutation of SerE306, LysE307 and ThrE332 was most likely attributable to loss of hydrogen bonding potential with E16, while ThrE330 appeared to stabilize the DIII N-terminal strand conformation and provided numerous van der Waals contacts with the Fab (FIGS. 13 and 14). Collectively, these structural studies defined the E16 epitope as a large surface patch on DIII created by four distinct secondary-structure elements, and the previously-performed yeast mapping highlighted the critical contributions of four central residues.
Comparison of available WNV sequences revealed nearly complete conservation of the structurally defined E16 epitope (FIG. 15). Not surprisingly, E16 blocked infection of ten different lineage I and II WNV strains. See Oliphant et al., 2005, Nat Med 11:522-30 (2005). Notably, nine other neutralizing mAbs also lost the ability to recognize DIII after mutation of SerE306, LysE307, IhrE330 or ThrE332. Importantly, this epitope is also key in the humoral immune response of humans, as E16 Fabs effectively competed WNV convalescent antibodies for DIII bindings. Sequence analysis of other flaviviruses reveals a high degree of diversity in the four segments of the E16 epitope, with notable variation even between dengue serotypes (FIG. 15). Not surprisingly, E16 did not cross-neutralize dengue, Japanese or St. Louis encephalitis viruses. Interestingly, other flavivirus-specific neutralizing antibodies localize to an analogous DIII binding region. See Wu et al., 2003, J. Biol. Chem. 278:46007-13; Volk et al., 2004, J. Biol. Chem. 279:38755-61; and Hiramatsu et al., 1996, Virology 224:437-45. Thus, the coincident mapping of mAbs that neutralize WNV and other flaviviruses suggested that the structural epitope recognized by E16 has a dominant role in flavivirus neutralization.
6.3 Superimposition of WNV D16-E16 Fab Structures onto Viral Structures
To gain additional insight into the structural basis of E16-mediated neutralization, the structure of the WNV DIII-E16 Fab complex was superimposed onto the structures of the pre-fusion dengue E dimer (see Modis et al., 2003, Proc Natl Acad Sci USA 100:6986-91 and post-fusion trimer (see Modis et al., 2004, Nature 427:313-9) (FIGS. 17 and 18, respectively). The E16 epitope was unencumbered in either configuration, although intact E16 IgG is unlikely to bivalently recognize these isolated oligomers due to extensive CHI domain splaying15 (FIGS. 17 and 18). DIII undergoes an  ̃70° rotation towards DII in the dimer to trimer transition (see Bressanelli et al., 2004, EMBO J. 23:728-38 and Modis et al., 2004, Nature 427:313-9), and E16 ligationper se could serve to hinder this conformational change. Moreover, E16 bound part of the linker that connects DIII to DI, and the N-terminus of truncated WNV DIII fragment adopts a unique conformation that enables TyrE302 to make contact with the E16 VH domain (FIG. 19). In E trimers, the flavivirus invariant TyrE302 interacts with DI in a manner that would be disrupted if re-oriented as observed in the E16/DIII complex. Thus, without intending to be bound to any particular theory or mechanism of action, E16 binding could stabilize the mature state or alternatively, restrict transition to the post-fusion conformation.
To better understand how E16 recognizes DIII in the context of the mature virus, the structure of WNV DIII-E16 Fab was superimposed onto the cryo-electron microscopy derived pseudo-atomic model of the intact WNV virion. See Mukhopadhyay et al., 2003, Science 302:248 and Kuhn et al., 2002, Cell 108:717-25. With three E proteins in the asymmetric unit, there were three potential Fab binding environments (FIG. 20). Two binding modes were clearly allowed: one that closely circles the three-fold axis and a second disposed symmetrically about the icosahedral dyad that was permuted as an outer five-fold ring (FIGS. 21A and 21B). However, the DIII epitopes were too tightly clustered at the true five-fold axis to permit E16 engagement without steric overlap with adjacent DIII residues. Thus, without intending to be bound to any particular theory or mechanism of action, it is believed that at saturation no more than 120 Fabs can bind the 180 E proteins in the mature virion, with exclusion of Fab binding to DIII around the inner five-fold ring (FIG. 22). Additional exclusions may occur for intact antibody, although it is noted that E16 Fab alone neutralizes WNV.
6.4 Functional Characterization of E16
Next, the ability of E16 to block cellular attachment of WNV was assessed. Binding assays were performed with Vero cells, a cell line permissive for WNV infection. After a 4° C. incubation with WNV in the presence of control (anti-SARS ORF7a), non-neutralizing (E22), or neutralizing mAbs that map within (E16 or E24) or outside (E53 or E60) of DIII, cell-associated viral RNA was measured by fluorogenic RT-PCR as described in Diamond et al., 2003, J. Virol. 77: 2578-2586. Briefly, individual purified mAbs (50 μg/ml of anti-SARS ORF7a, E16, E22, E24, E53, or E60) or medium alone were incubated with 103 PFU of WNV for one hour at 4° C. These virus-antibody mixtures were then added to Vero cells in 12-well plates for one hour on ice. Unbound virus was removed after six washes with PBS at 4° C. Cells were lysed with guanidinium isothiocyanate, RNA was purified, and viral RNA was quantified by fluorogenic RT-PCR18.
Importantly, the non-binding and non-neutralizing mAbs did not inhibit virus binding. In contrast, E53 and E60 blocked virus attachment by 8 to 9 fold (P<0.001) whereas E16 and E24, which recognize the same dominant DIII epitope, only inhibited binding by 3.5-fold (P=0.003) (FIG. 23). The observation that E53 and E60 blocked virus binding more efficiently than E16 was not expected, as E53 and E60 were tenfold less potent in plaque reduction neutralization assays. See Oliphant et al., 2005, Nat. Med. 11:522-30.
Because E16 only partially blocked virus binding yet completely neutralized infection, whether E16 inhibits flavivirus infection by blocking a step after cellular attachment was next assessed. Using a previously described assay, E16 or E53 was incubated with WNV prior to, or after, mixing with a monolayer of Vero cells and infection was measured as described in Crill & Roehrig J, 2001, J. Virol. 75:7769-73 and Hung et al., 1999, Virology 257:156-67. Briefly, increasing concentrations of E16 or E53 were added prior to or after WNV (102 PFU) binding (one hour on ice) to Vero cells. In the post-adsorption assay, after washing away unbound virus, mAb was allowed to bind for an additional hour. All cells were washed and an agarose overlay was added. Three days later, plaques were scored after fixation and staining with crystal violet.
Pre-binding of WNV with either E16 or E53 significantly protected against infection (FIG. 24). In contrast, E16 but not E53 significantly inhibited infection when added after virus binding. Because E16-mediated protection was not appreciably affected by the time of addition, it is believed that it acts primarily after WNV cellular attachment.
To further define the mechanism of WNV neutralization, the ability of E16 or other mAbs to enhance infection in macrophages was evaluated. Antibody-dependent enhancement of infection occurs when antibody/virus complexes are preferentially internalized through Fcγ receptors on myeloid cells. Although the in vivo consequences remain uncertain, many mAbs efficiently enhance flavivirus infection of Fcγ receptor bearing cells even when inhibitory in fibroblast neutralization assays. See Halstead & O'Rourke, 1977, Nature 265:739-41. The ability of saturating concentrations of non-neutralizing (E5) or neutralizing (E16, E24, or E60) mAbs to enhance WNV infection was therefore assessed in macrophages. Briefly, WNV (5×102 PFU) was pre-incubated with media, individual mAbs (50 μg/ml of E5, E16, E24 or E60) or combinations of mAbs (E16+E5, E60+E5, or E16+E60, E24+E5, E24+E60) and then added to a monolayer (105) of J774.2 murine macrophages. After 6 hours, cells were washed extensively with PBS to remove unbound virus and rnAb. After an additional 24 hours, supernatants were harvested for a viral plaque assay on Vero cells.
While E5 and E60 augmented infection 270 and 3,000-fold respectively, E16 potently inhibited macrophage infection at the same concentration. Strikingly, when E16 is combined with E5 or E60, it completely blocks enhancement as judged by reduction of virus yield (FIG. 25) or viral RNA. E24, which maps to the E16 dominant epitope, also blocked E5 and E60-dependent enhancement. Finally, the blockade of enhancement was not due to epitope competition as E16 and E24 do not cross-compete E5 or E60 for WNV E binding (FIG. 26). To perform the competition experiments, yeast expressing the empty vector pY D 1 or the WNV E ectodomain (residues 1-415) were incubated with 2.5 μg unlabeled E16 or E60 antibody for one hour on ice. Unbound antibody was removed after three PBS washes containing 1 mg/ml BSA. E16 and E60 were conjugated using an Alexa Flour 647 Monoclonal Antibody Labeling Kit (Molecular Probes). Conjugated E16 or E60 (25 μg/ml) was then added to the cells for 30 minutes at 4° C. Yeasts were washed three times with PBS, fixed with 1% paraformaldehyde, and analyzed using flow cytometry.
Collectively, these virologic experiments strongly suggest E16 blocked WNV infection primarily after cellular attachment.
In summary, E16 binds a dominant neutralizing epitope on WNV E defined by four distinct secondary-structure elements that create a large surface patch on DIII, a region associated with pH-dependent conformational changes. Modeling studies suggest that E16 was excluded from five-fold clustered DIII epitopes on mature virions, potentially leaving them free to serve in receptor binding. Consistent with this, E16 inhibits WNV infection primarily at a step after virus attachment. Moreover, these data suggest a potential advantage for interfering with post-attachment events
The present invention is not to be limited in scope by the exemplified embodiments, which are intended as illustrations of single aspects of the invention. Indeed, various modifications of the invention in addition to those described herein will become apparent to those having skill in the art from the foregoing description and accompanying drawings. Such modifications are intended to fall with in the scope of the appended claims.
All documents referenced in this application, whether patents, published or unpublished patent applications, either U.S. or foreign, literature references, nucleotide or amino acid sequences identified by Accession No. or otherwise, are hereby incorporated by reference in their entireties for any and all purposes.


















TABLE 7







ATOM
1
CB
ASP
L
1
−35.475
10.622
12.124
1
30.78
L


ATOM
2
CG
ASP
L
1
−34.677
11.822
11.637
1
33.43
L


ATOM
3
OD1
ASP
L
1
−35.196
12.58
10.789
1
33.95
L


ATOM
4
OD2
ASP
L
1
−33.529
12.007
12.093
1
37.37
L


ATOM
5
C
ASP
L
1
−37.648
9.792
13.041
1
30.81
L


ATOM
6
O
ASP
L
1
−38.023
9.855
14.209
1
34.21
L


ATOM
7
N
ASP
L
1
−37.02
12.125
13.334
1
28.93
L


ATOM
8
CA
ASP
L
1
−36.938
10.97
12.402
1
30.33
L


ATOM
9
N
ILE
L
2
−37.837
8.713
12.292
1
28.58
L


ATOM
10
CA
ILE
L
2
−38.533
7.567
12.852
1
27.42
L


ATOM
11
CB
ILE
L
2
−39.122
6.655
11.743
1
28.5
L


ATOM
12
CG2
ILE
L
2
−39.914
5.507
12.373
1
23.83
L


ATOM
13
CG1
ILE
L
2
−40.05
7.483
10.843
1
28.97
L


ATOM
14
CD1
ILE
L
2
−40.839
6.681
9.836
1
31.74
L


ATOM
15
C
ILE
L
2
−37.625
6.764
13.769
1
26.09
L


ATOM
16
O
ILE
L
2
−36.557
6.313
13.377
1
24.34
L


ATOM
17
N
VAL
L
3
−38.073
6.603
15.007
1
26.39
L


ATOM
18
CA
VAL
L
3
−37.324
5.878
16.017
1
24.96
L


ATOM
19
CB
VAL
L
3
−37.489
6.563
17.393
1
24.46
L


ATOM
20
CG1
VAL
L
3
−36.764
5.776
18.476
1
26.77
L


ATOM
21
CG2
VAL
L
3
−36.933
7.966
17.321
1
22.59
L


ATOM
22
C
VAL
L
3
−37.774
4.426
16.096
1
23.93
L


ATOM
23
O
VAL
L
3
−38.955
4.139
16.27
1
24.37
L


ATOM
24
N
MET
L
4
−36.818
3.517
15.959
1
23.88
L


ATOM
25
CA
MET
L
4
−37.1
2.094
16.01
1
26.48
L


ATOM
26
CB
MET
L
4
−36.377
1.361
14.875
1
26.68
L


ATOM
27
CG
MET
L
4
−36.771
1.83
13.485
1
27.04
L


ATOM
28
SD
MET
L
4
−38.549
1.793
13.233
1
29.49
L


ATOM
29
CE
MET
L
4
−38.785
0.088
12.812
1
25.37
L


ATOM
30
C
MET
L
4
−36.666
1.516
17.344
1
28.26
L


ATOM
31
O
MET
L
4
−35.477
1.486
17.677
1
29.27
L


ATOM
32
N
THR
L
5
−37.639
1.043
18.105
1
28.06
L


ATOM
33
CA
THR
L
5
−37.343
0.47
19.4
1
30.04
L


ATOM
34
CB
THR
L
5
−38.063
1.247
20.512
1
30.06
L


ATOM
35
OG1
THR
L
5
−38.025
0.488
21.726
1
30.53
L


ATOM
36
CG2
THR
L
5
−39.503
1.512
20.125
1
32.13
L


ATOM
37
C
THR
L
5
−37.74
−0.996
19.454
1
27.75
L


ATOM
38
O
THR
L
5
−38.903
−1.341
19.262
1
26.81
L


ATOM
39
N
GLN
L
6
−36.755
−1.85
19.717
1
27.96
L


ATOM
40
CA
GLN
L
6
−36.967
−3.293
19.802
1
28.11
L


ATOM
41
CB
GLN
L
6
−35.754
−4.025
19.233
1
24.74
L


ATOM
42
CG
GLN
L
6
−35.445
−3.659
17.8
1
26.48
L


ATOM
43
CD
GLN
L
6
−34.369
−4.54
17.203
1
29.46
L


ATOM
44
OE1
GLN
L
6
−33.968
−5.54
17.802
1
30.77
L


ATOM
45
NE2
GLN
L
6
−33.902
−4.184
16.012
1
30.91
L


ATOM
46
C
GLN
L
6
−37.212
−3.767
21.238
1
28.49
L


ATOM
47
O
GLN
L
6
−36.892
−3.061
22.201
1
24.09
L


ATOM
48
N
SER
L
7
−37.785
−4.961
21.374
1
28.74
L


ATOM
49
CA
SER
L
7
−38.046
−5.521
22.696
1
35.25
L


ATOM
50
CB
SER
L
7
−38.811
−6.853
22.581
1
34.58
L


ATOM
51
OG
SER
L
7
−38.283
−7.686
21.559
1
37.01
L


ATOM
52
C
SER
L
7
−36.707
−5.716
23.417
1
36.8
L


ATOM
53
O
SER
L
7
−35.652
−5.705
22.776
1
38.22
L


ATOM
54
N
HIS
L
8
−36.753
−5.892
24.735
1
37.65
L


ATOM
55
CA
HIS
L
8
−35.539
−6.051
25.539
1
41.93
L


ATOM
56
CB
HIS
L
8
−35.853
−6.806
26.843
1
47.76
L


ATOM
57
CG
HIS
L
8
−36.614
−8.082
26.651
1
53.99
L


ATOM
58
CD2
HIS
L
8
−37.855
−8.317
26.163
1
56.53
L


ATOM
59
ND1
HIS
L
8
−36.111
−9.311
27.028
1
56.37
L


ATOM
60
CE1
HIS
L
8
−37.01
−10.247
26.781
1
56.96
L


ATOM
61
NE2
HIS
L
8
−38.077
−9.67
26.256
1
59.42
L


ATOM
62
C
HIS
L
8
−34.346
−6.688
24.821
1
40.37
L


ATOM
63
O
HIS
L
8
−34.409
−7.835
24.373
1
41.01
L


ATOM
64
N
LYS
L
9
−33.259
−5.918
24.738
1
37.22
L


ATOM
65
CA
LYS
L
9
−32.022
−6.315
24.065
1
37.72
L


ATOM
66
CB
LYS
L
9
−30.957
−5.237
24.266
1
36.46
L


ATOM
67
CG
LYS
L
9
−29.714
−5.454
23.428
1
37.92
L


ATOM
68
CD
LYS
L
9
−28.909
−4.178
23.31
1
41.81
L


ATOM
69
CE
LYS
L
9
−27.874
−4.282
22.196
1
43.88
L


ATOM
70
NZ
LYS
L
9
−27.162
−2.987
21.979
1
45.07
L


ATOM
71
C
LYS
L
9
−31.412
−7.671
24.418
1
38.69
L


ATOM
72
O
LYS
L
9
−30.833
−8.335
23.559
1
40.24
L


ATOM
73
N
PHE
L
10
−31.51
−8.083
25.675
1
38.73
L


ATOM
74
CA
PHE
L
10
−30.938
−9.364
26.059
1
38.54
L


ATOM
75
CB
PHE
L
10
−29.949
−9.183
27.208
1
35.83
L


ATOM
76
CG
PHE
L
10
−28.768
−8.339
26.847
1
31.3
L


ATOM
77
CD1
PHE
L
10
−28.872
−6.951
26.82
1
28.99
L


ATOM
78
CD2
PHE
L
10
−27.562
−8.935
26.485
1
29.58
L


ATOM
79
CE1
PHE
L
10
−27.791
−6.162
26.434
1
29.51
L


ATOM
80
CE2
PHE
L
10
−26.469
−8.159
26.096
1
29.5
L


ATOM
81
CZ
PHE
L
10
−26.584
−6.765
26.071
1
28.97
L


ATOM
82
C
PHE
L
10
−32.011
−10.363
26.436
1
41.36
L


ATOM
83
O
PHE
L
10
−32.318
−10.563
27.616
1
41.31
L


ATOM
84
N
MET
L
11
−32.582
−10.989
25.414
1
42.37
L


ATOM
85
CA
MET
L
11
−33.625
−11.971
25.625
1
42.65
L


ATOM
86
CB
MET
L
11
−34.709
−11.815
24.558
1
45.16
L


ATOM
87
CG
MET
L
11
−34.302
−12.238
23.161
1
48.29
L


ATOM
88
SD
MET
L
11
−34.879
−13.901
22.774
1
52.68
L


ATOM
89
CE
MET
L
11
−33.388
−14.833
22.912
1
51.29
L


ATOM
90
C
MET
L
11
−33.034
−13.373
25.589
1
41.43
L


ATOM
91
O
MET
L
11
−31.969
−13.595
25.01
1
41.95
L


ATOM
92
N
SER
L
12
−33.727
−14.314
26.221
1
39.4
L


ATOM
93
CA
SER
L
12
−33.272
−15.696
26.268
1
36.95
L


ATOM
94
CB
SER
L
12
−32.894
−16.08
27.696
1
35.26
L


ATOM
95
OG
SER
L
12
−31.886
−15.223
28.192
1
43.53
L


ATOM
96
C
SER
L
12
−34.344
−16.648
25.772
1
34.68
L


ATOM
97
O
SER
L
12
−35.517
−16.301
25.715
1
33.89
L


ATOM
98
N
THR
L
13
−33.927
−17.854
25.414
1
33.57
L


ATOM
99
CA
THR
L
13
−34.846
−18.874
24.944
1
34.81
L


ATOM
100
CB
THR
L
13
−35.316
−18.6
23.506
1
36.65
L


ATOM
101
OG1
THR
L
13
−36.301
−19.574
23.127
1
37.88
L


ATOM
102
CG2
THR
L
13
−34.139
−18.668
22.54
1
34.9
L


ATOM
103
C
THR
L
13
−34.104
−20.193
24.969
1
35.21
L


ATOM
104
O
THR
L
13
−32.891
−20.224
25.135
1
35.62
L


ATOM
105
N
SER
L
14
−34.835
−21.282
24.807
1
34.54
L


ATOM
106
CA
SER
L
14
−34.22
−22.593
24.804
1
36.72
L


ATOM
107
CB
SER
L
14
−34.931
−23.488
25.81
1
38.64
L


ATOM
108
OG
SER
L
14
−36.332
−23.371
25.666
1
43.39
L


ATOM
109
C
SER
L
14
−34.317
−23.181
23.405
1
37.15
L


ATOM
110
O
SER
L
14
−35.317
−22.996
22.715
1
38.61
L


ATOM
111
N
VAL
L
15
−33.272
−23.88
22.982
1
35.82
L


ATOM
112
CA
VAL
L
15
−33.264
−24.482
21.658
1
36.21
L


ATOM
113
CB
VAL
L
15
−32.116
−25.511
21.524
1
36.7
L


ATOM
114
CG1
VAL
L
15
−32.292
−26.344
20.259
1
34.02
L


ATOM
115
CG2
VAL
L
15
−30.778
−24.784
21.493
1
37.6
L


ATOM
116
C
VAL
L
15
−34.587
−25.176
21.363
1
35.63
L


ATOM
117
O
VAL
L
15
−35.11
−25.902
22.199
1
33.85
L


ATOM
118
N
GLY
L
16
−35.127
−24.935
20.172
1
36.61
L


ATOM
119
CA
GLY
L
16
−36.375
−25.564
19.779
1
37.2
L


ATOM
120
C
GLY
L
16
−37.617
−24.763
20.097
1
37.26
L


ATOM
121
O
GLY
L
16
−38.681
−25.004
19.53
1
38.61
L


ATOM
122
N
ASP
L
17
−37.495
−23.809
21.006
1
38.32
L


ATOM
123
CA
ASP
L
17
−38.638
−22.987
21.364
1
39.24
L


ATOM
124
CB
ASP
L
17
−38.421
−22.332
22.727
1
41.91
L


ATOM
125
CG
ASP
L
17
−39.453
−22.766
23.736
1
45.3
L


ATOM
126
OD1
ASP
L
17
−40.656
−22.541
23.485
1
46.3
L


ATOM
127
OD2
ASP
L
17
−39.068
−23.341
24.773
1
48.62
L


ATOM
128
C
ASP
L
17
−38.909
−21.913
20.321
1
38.09
L


ATOM
129
O
ASP
L
17
−38.175
−21.774
19.342
1
34.63
L


ATOM
130
N
ARG
L
18
−39.982
−21.165
20.54
1
38.88
L


ATOM
131
CA
ARG
L
18
−40.369
−20.091
19.644
1
40.46
L


ATOM
132
CB
ARG
L
18
−41.862
−20.17
19.325
1
43.7
L


ATOM
133
CG
ARG
L
18
−42.402
−21.568
19.113
1
49.69
L


ATOM
134
CD
ARG
L
18
−43.843
−21.509
18.635
1
55.22
L


ATOM
135
NE
ARG
L
18
−43.943
−20.903
17.306
1
60.22
L


ATOM
136
CZ
ARG
L
18
−45.075
−20.78
16.617
1
61.55
L


ATOM
137
NH1
ARG
L
18
−46.22
−21.217
17.128
1
62.33
L


ATOM
138
NH2
ARG
L
18
−45.061
−20.234
15.406
1
60.76
L


ATOM
139
C
ARG
L
18
−40.097
−18.78
20.366
1
39.91
L


ATOM
140
O
ARG
L
18
−40.417
−18.643
21.542
1
41.43
L


ATOM
141
N
VAL
L
19
−39.5
−17.82
19.675
1
37.42
L


ATOM
142
CA
VAL
L
19
−39.24
−16.526
20.289
1
35.66
L


ATOM
143
CB
VAL
L
19
−37.743
−16.284
20.528
1
35.42
L


ATOM
144
CG1
VAL
L
19
−37.179
−17.419
21.333
1
39.42
L


ATOM
145
CG2
VAL
L
19
−37.012
−16.136
19.202
1
32.76
L


ATOM
146
C
VAL
L
19
−39.751
−15.438
19.375
1
33.44
L


ATOM
147
O
VAL
L
19
−40.015
−15.682
18.195
1
33.86
L


ATOM
148
N
SER
L
20
−39.89
−14.239
19.927
1
30.78
L


ATOM
149
CA
SER
L
20
−40.351
−13.095
19.151
1
29.79
L


ATOM
150
CB
SER
L
20
−41.865
−12.903
19.315
1
27
L


ATOM
151
OG
SER
L
20
−42.587
−14.032
18.867
1
22.54
L


ATOM
152
C
SER
L
20
−39.637
−11.81
19.563
1
28.33
L


ATOM
153
O
SER
L
20
−39.775
−11.339
20.696
1
27.37
L


ATOM
154
N
ILE
L
21
−38.849
−11.256
18.651
1
27.04
L


ATOM
155
CA
ILE
L
21
−38.182
−9.999
18.942
1
26.07
L


ATOM
156
CB
ILE
L
21
−36.828
−9.839
18.228
1
23.33
L


ATOM
157
CG2
ILE
L
21
−36.203
−8.518
18.626
1
23.97
L


ATOM
158
CG1
ILE
L
21
−35.888
−10.982
18.59
1
23.16
L


ATOM
159
CD1
ILE
L
21
−36.186
−12.255
17.85
1
26.47
L


ATOM
160
C
ILE
L
21
−39.141
−9.012
18.325
1
25.95
L


ATOM
161
O
ILE
L
21
−39.53
−9.174
17.169
1
25.34
L


ATOM
162
N
THR
L
22
−39.551
−8.007
19.082
1
24.86
L


ATOM
163
CA
THR
L
22
−40.472
−7.043
18.52
1
25.78
L


ATOM
164
CB
THR
L
22
−41.627
−6.748
19.483
1
26.92
L


ATOM
165
OG1
THR
L
22
−41.108
−6.381
20.766
1
30.07
L


ATOM
166
CG2
THR
L
22
−42.51
−7.974
19.614
1
26.84
L


ATOM
167
C
THR
L
22
−39.773
−5.757
18.138
1
24.77
L


ATOM
168
O
THR
L
22
−38.652
−5.487
18.566
1
22.7
L


ATOM
169
N
CYS
L
23
−40.453
−4.969
17.321
1
25.79
L


ATOM
170
CA
CYS
L
23
−39.917
−3.713
16.843
1
28.58
L


ATOM
171
C
CYS
L
23
−41.051
−2.707
16.786
1
28.73
L


ATOM
172
O
CYS
L
23
−42.083
−2.962
16.17
1
28.61
L


ATOM
173
CB
CYS
L
23
−39.332
−3.933
15.448
1
31.5
L


ATOM
174
SG
CYS
L
23
−38.547
−2.535
14.577
1
35.43
L


ATOM
175
N
LYS
L
24
−40.864
−1.571
17.442
1
29.33
L


ATOM
176
CA
LYS
L
24
−41.872
−0.526
17.43
1
32.95
L


ATOM
177
CB
LYS
L
24
−42.305
−0.16
18.854
1
38.59
L


ATOM
178
CG
LYS
L
24
−43.375
0.933
18.893
1
45.28
L


ATOM
179
CD
LYS
L
24
−43.655
1.411
20.312
1
50.68
L


ATOM
180
CE
LYS
L
24
−44.83
2.387
20.349
1
52.5
L


ATOM
181
NZ
LYS
L
24
−46.127
1.738
19.996
1
52.23
L


ATOM
182
C
LYS
L
24
−41.309
0.713
16.756
1
30.25
L


ATOM
183
O
LYS
L
24
−40.172
1.099
17.011
1
32.95
L


ATOM
184
N
ALA
L
25
−42.105
1.329
15.891
1
29.22
L


ATOM
185
CA
ALA
L
25
−41.68
2.542
15.207
1
27.96
L


ATOM
186
CB
ALA
L
25
−41.877
2.405
13.708
1
26.41
L


ATOM
187
C
ALA
L
25
−42.524
3.683
15.753
1
26.47
L


ATOM
188
O
ALA
L
25
−43.712
3.513
15.999
1
21.99
L


ATOM
189
N
SER
L
26
−41.9
4.841
15.949
1
28.43
L


ATOM
190
CA
SER
L
26
−42.589
6.016
16.479
1
29.22
L


ATOM
191
CB
SER
L
26
−41.592
7.17
16.626
1
27.56
L


ATOM
192
OG
SER
L
26
−40.8
7.32
15.46
1
28.74
L


ATOM
193
C
SER
L
26
−43.819
6.452
15.657
1
29.46
L


ATOM
194
O
SER
L
26
−44.611
7.282
16.106
1
26.09
L


ATOM
195
N
GLN
L
27
−43.972
5.885
14.461
1
30.97
L


ATOM
196
CA
GLN
L
27
−45.116
6.179
13.6
1
31.29
L


ATOM
197
CB
GLN
L
27
−44.906
7.491
12.825
1
34.18
L


ATOM
198
CG
GLN
L
27
−43.578
7.639
12.109
1
36.72
L


ATOM
199
CD
GLN
L
27
−43.439
8.999
11.437
1
38.14
L


ATOM
200
OE1
GLN
L
27
−44.176
9.325
10.503
1
38.97
L


ATOM
201
NE2
GLN
L
27
−42.496
9.802
11.915
1
38.77
L


ATOM
202
C
GLN
L
27
−45.379
5.023
12.639
1
30.09
L


ATOM
203
O
GLN
L
27
−44.548
4.122
12.509
1
29.44
L


ATOM
204
N
ASP
L
28
−46.543
5.043
11.991
1
27.3
L


ATOM
205
CA
ASP
L
28
−46.932
3.994
11.05
1
25.1
L


ATOM
206
CB
ASP
L
28
−48.333
4.283
10.501
1
24.08
L


ATOM
207
CG
ASP
L
28
−48.934
3.099
9.75
1
29.29
L


ATOM
208
OD1
ASP
L
28
−48.175
2.208
9.312
1
32.01
L


ATOM
209
OD2
ASP
L
28
−50.176
3.062
9.583
1
28.91
L


ATOM
210
C
ASP
L
28
−45.926
3.96
9.901
1
26.55
L


ATOM
211
O
ASP
L
28
−45.831
4.912
9.128
1
27.44
L


ATOM
212
N
VAL
L
29
−45.161
2.877
9.797
1
25.47
L


ATOM
213
CA
VAL
L
29
−44.178
2.751
8.722
1
25.32
L


ATOM
214
CB
VAL
L
29
−42.846
2.171
9.234
1
26.06
L


ATOM
215
CG1
VAL
L
29
−42.018
3.288
9.867
1
23.41
L


ATOM
216
CG2
VAL
L
29
−43.116
1.056
10.244
1
21.47
L


ATOM
217
C
VAL
L
29
−44.726
1.866
7.612
1
24.8
L


ATOM
218
O
VAL
L
29
−43.99
1.18
6.906
1
24.35
L


ATOM
219
N
SER
L
30
−46.047
1.907
7.487
1
26.52
L


ATOM
220
CA
SER
L
30
−46.809
1.172
6.492
1
26
L


ATOM
221
CB
SER
L
30
−47.044
2.083
5.291
1
24.48
L


ATOM
222
OG
SER
L
30
−48.007
1.526
4.421
1
31.89
L


ATOM
223
C
SER
L
30
−46.255
−0.184
6.023
1
26.66
L


ATOM
224
O
SER
L
30
−46.06
−0.401
4.821
1
23.27
L


ATOM
225
N
THR
L
31
−46.003
−1.095
6.964
1
25.31
L


ATOM
226
CA
THR
L
31
−45.513
−2.423
6.599
1
25.99
L


ATOM
227
CB
THR
L
31
−46.478
−3.045
5.539
1
25.05
L


ATOM
228
OG1
THR
L
31
−47.553
−3.698
6.224
1
25.02
L


ATOM
229
CG2
THR
L
31
−45.775
−4.02
4.618
1
21.63
L


ATOM
230
C
THR
L
31
−44.068
−2.463
6.092
1
25.19
L


ATOM
231
O
THR
L
31
−43.49
−3.537
5.919
1
24.76
L


ATOM
232
N
ALA
L
32
−43.479
−1.292
5.871
1
24.51
L


ATOM
233
CA
ALA
L
32
−42.111
−1.214
5.371
1
23.42
L


ATOM
234
CB
ALA
L
32
−41.902
0.115
4.65
1
25.37
L


ATOM
235
C
ALA
L
32
−41.052
−1.392
6.459
1
20.82
L


ATOM
236
O
ALA
L
32
−40.422
−0.427
6.888
1
19.59
L


ATOM
237
N
VAL
L
33
−40.852
−2.631
6.897
1
20.92
L


ATOM
238
CA
VAL
L
33
−39.861
−2.927
7.927
1
19.82
L


ATOM
239
CB
VAL
L
33
−40.521
−3.096
9.336
1
20.11
L


ATOM
240
CG1
VAL
L
33
−39.538
−3.724
10.325
1
19.01
L


ATOM
241
CG2
VAL
L
33
−40.951
−1.755
9.865
1
18.53
L


ATOM
242
C
VAL
L
33
−39.1
−4.202
7.57
1
20.5
L


ATOM
243
O
VAL
L
33
−39.678
−5.186
7.091
1
17.51
L


ATOM
244
N
ALA
L
34
−37.795
−4.174
7.809
1
18.68
L


ATOM
245
CA
ALA
L
34
−36.944
−5.316
7.515
1
18.72
L


ATOM
246
CB
ALA
L
34
−35.957
−4.949
6.417
1
12.5
L


ATOM
247
C
ALA
L
34
−36.188
−5.795
8.757
1
20.05
L


ATOM
248
O
ALA
L
34
−36.031
−5.057
9.739
1
19.38
L


ATOM
249
N
TRP
L
35
−35.729
−7.039
8.712
1
17.67
L


ATOM
250
CA
TRP
L
35
−34.968
−7.586
9.815
1
17.47
L


ATOM
251
CB
TRP
L
35
−35.759
−8.67
10.533
1
17.5
L


ATOM
252
CG
TRP
L
35
−36.962
−8.18
11.266
1
21.69
L


ATOM
253
CD2
TRP
L
35
−37.034
−7.833
12.653
1
21.78
L


ATOM
254
CE2
TRP
L
35
−38.374
−7.476
12.924
1
22.24
L


ATOM
255
CE3
TRP
L
35
−36.094
−7.788
13.695
1
22.95
L


ATOM
256
CD1
TRP
L
35
−38.225
−8.018
10.763
1
20.24
L


ATOM
257
NE1
TRP
L
35
−39.076
−7.601
11.754
1
21.01
L


ATOM
258
CZ2
TRP
L
35
−38.803
−7.079
14.198
1
23.86
L


ATOM
259
CZ3
TRP
L
35
−36.517
−7.391
14.964
1
25.12
L


ATOM
260
CH2
TRP
L
35
−37.863
−7.041
15.203
1
25.78
L


ATOM
261
C
TRP
L
35
−33.651
−8.167
9.328
1
15.83
L


ATOM
262
O
TRP
L
35
−33.603
−8.86
8.323
1
15.93
L


ATOM
263
N
TYR
L
36
−32.577
−7.882
10.049
1
16.64
L


ATOM
264
CA
TYR
L
36
−31.271
−8.404
9.679
1
21.5
L


ATOM
265
CB
TYR
L
36
−30.285
−7.267
9.379
1
19.96
L


ATOM
266
CG
TYR
L
36
−30.714
−6.347
8.261
1
21.18
L


ATOM
267
CD1
TYR
L
36
−30.503
−6.688
6.925
1
20.53
L


ATOM
268
CE1
TYR
L
36
−30.89
−5.835
5.893
1
19.8
L


ATOM
269
CD2
TYR
L
36
−31.329
−5.126
8.543
1
22.81
L


ATOM
270
CE2
TYR
L
36
−31.726
−4.263
7.522
1
20.1
L


ATOM
271
CZ
TYR
L
36
−31.504
−4.621
6.201
1
21.52
L


ATOM
272
OH
TYR
L
36
−31.9
−3.764
5.198
1
18.17
L


ATOM
273
C
TYR
L
36
−30.739
−9.218
10.842
1
21.62
L


ATOM
274
O
TYR
L
36
−31.117
−8.993
11.992
1
21.98
L


ATOM
275
N
GLN
L
37
−29.862
−10.164
10.528
1
22.38
L


ATOM
276
CA
GLN
L
37
−29.23
−11.007
11.53
1
21.58
L


ATOM
277
CB
GLN
L
37
−29.485
−12.488
11.24
1
22.59
L


ATOM
278
CG
GLN
L
37
−28.769
−13.432
12.192
1
20.06
L


ATOM
279
CD
GLN
L
37
−28.824
−14.868
11.73
1
24.01
L


ATOM
280
OE1
GLN
L
37
−28.373
−15.194
10.63
1
24.4
L


ATOM
281
NE2
GLN
L
37
−29.377
−15.742
12.566
1
24.16
L


ATOM
282
C
GLN
L
37
−27.736
−10.739
11.453
1
22.35
L


ATOM
283
O
GLN
L
37
−27.155
−10.709
10.372
1
19.14
L


ATOM
284
N
GLN
L
38
−27.107
−10.52
12.597
1
26.93
L


ATOM
285
CA
GLN
L
38
−25.678
−10.286
12.582
1
28.65
L


ATOM
286
CB
GLN
L
38
−25.362
−8.817
12.815
1
28.44
L


ATOM
287
CG
GLN
L
38
−23.879
−8.555
12.826
1
30.45
L


ATOM
288
CD
GLN
L
38
−23.55
−7.093
12.869
1
31.99
L


ATOM
289
OE1
GLN
L
38
−24.099
−6.336
13.681
1
33.27
L


ATOM
290
NE2
GLN
L
38
−22.638
−6.677
11.999
1
32.4
L


ATOM
291
C
GLN
L
38
−24.953
−11.141
13.6
1
29.69
L


ATOM
292
O
GLN
L
38
−25.154
−11.008
14.806
1
28.23
L


ATOM
293
N
LYS
L
39
−24.116
−12.034
13.084
1
33.97
L


ATOM
294
CA
LYS
L
39
−23.324
−12.935
13.907
1
36.3
L


ATOM
295
CB
LYS
L
39
−23.229
−14.306
13.231
1
33.95
L


ATOM
296
CG
LYS
L
39
−24.566
−15.002
13.168
1
32.64
L


ATOM
297
CD
LYS
L
39
−24.509
−16.344
12.469
1
32.98
L


ATOM
298
CE
LYS
L
39
−25.863
−17.034
12.602
1
31.48
L


ATOM
299
NZ
LYS
L
39
−25.98
−18.286
11.819
1
35.86
L


ATOM
300
C
LYS
L
39
−21.938
−12.334
14.107
1
38.02
L


ATOM
301
O
LYS
L
39
−21.411
−11.656
13.223
1
37.92
L


ATOM
302
N
PRO
L
40
−21.336
−12.572
15.281
1
38.79
L


ATOM
303
CD
PRO
L
40
−21.857
−13.483
16.315
1
39.44
L


ATOM
304
CA
PRO
L
40
−20.009
−12.075
15.652
1
39.28
L


ATOM
305
CB
PRO
L
40
−19.593
−13.033
16.756
1
40.48
L


ATOM
306
CG
PRO
L
40
−20.881
−13.269
17.46
1
41.08
L


ATOM
307
C
PRO
L
40
−19.005
−12.036
14.502
1
38.95
L


ATOM
308
O
PRO
L
40
−18.731
−13.057
13.865
1
36.36
L


ATOM
309
N
GLY
L
41
−18.468
−10.844
14.247
1
38.48
L


ATOM
310
CA
GLY
L
41
−17.497
−10.67
13.182
1
38.13
L


ATOM
311
C
GLY
L
41
−18.038
−11.086
11.831
1
38.14
L


ATOM
312
O
GLY
L
41
−17.392
−11.831
11.098
1
39.38
L


ATOM
313
N
GLN
L
42
−19.235
−10.608
11.508
1
37.31
L


ATOM
314
CA
GLN
L
42
−19.876
−10.917
10.238
1
34.17
L


ATOM
315
CB
GLN
L
42
−20.751
−12.16
10.364
1
34.42
L


ATOM
316
CG
GLN
L
42
−19.988
−13.453
10.31
1
36.12
L


ATOM
317
CD
GLN
L
42
−20.896
−14.647
10.086
1
39.96
L


ATOM
318
OE1
GLN
L
42
−21.833
−14.594
9.275
1
37.78
L


ATOM
319
NE2
GLN
L
42
−20.617
−15.74
10.791
1
40.29
L


ATOM
320
C
GLN
L
42
−20.73
−9.751
9.781
1
34.27
L


ATOM
321
O
GLN
L
42
−21.189
−8.942
10.589
1
35.05
L


ATOM
322
N
SER
L
43
−20.94
−9.661
8.477
1
32.51
L


ATOM
323
CA
SER
L
43
−21.756
−8.59
7.938
1
30.71
L


ATOM
324
CB
SER
L
43
−21.585
−8.512
6.421
1
33.35
L


ATOM
325
OG
SER
L
43
−20.217
−8.381
6.074
1
40.08
L


ATOM
326
C
SER
L
43
−23.202
−8.923
8.265
1
28.28
L


ATOM
327
O
SER
L
43
−23.544
−10.085
8.471
1
28.57
L


ATOM
328
N
PRO
L
44
−24.068
−7.906
8.345
1
24.68
L


ATOM
329
CD
PRO
L
44
−23.796
−6.46
8.39
1
21.7
L


ATOM
330
CA
PRO
L
44
−25.47
−8.181
8.643
1
20.8
L


ATOM
331
CB
PRO
L
44
−26.089
−6.799
8.66
1
19.34
L


ATOM
332
CG
PRO
L
44
−24.985
−5.946
9.149
1
19.68
L


ATOM
333
C
PRO
L
44
−26.017
−9.025
7.503
1
24.4
L


ATOM
334
O
PRO
L
44
−25.49
−8.977
6.388
1
25.24
L


ATOM
335
N
LYS
L
45
−27.057
−9.808
7.78
1
27.66
L


ATOM
336
CA
LYS
L
45
−27.681
−10.644
6.752
1
27.45
L


ATOM
337
CB
LYS
L
45
−27.445
−12.132
7.025
1
28.38
L


ATOM
338
CG
LYS
L
45
−25.973
−12.507
7.118
1
35.81
L


ATOM
339
CD
LYS
L
45
−25.189
−11.936
5.939
1
40.78
L


ATOM
340
CE
LYS
L
45
−23.697
−12.201
6.061
1
41.63
L


ATOM
341
NZ
LYS
L
45
−23.408
−13.66
6.047
1
46.51
L


ATOM
342
C
LYS
L
45
−29.169
−10.354
6.731
1
26.01
L


ATOM
343
O
LYS
L
45
−29.827
−10.358
7.774
1
25.82
L


ATOM
344
N
LEU
L
46
−29.696
−10.09
5.543
1
22.57
L


ATOM
345
CA
LEU
L
46
−31.113
−9.788
5.404
1
24.84
L


ATOM
346
CB
LEU
L
46
−31.396
−9.282
3.992
1
24.51
L


ATOM
347
CG
LEU
L
46
−32.851
−8.956
3.692
1
23.39
L


ATOM
348
CD1
LEU
L
46
−33.306
−7.822
4.583
1
25.7
L


ATOM
349
CD2
LEU
L
46
−32.993
−8.581
2.228
1
25.19
L


ATOM
350
C
LEU
L
46
−31.984
−11.007
5.703
1
22.66
L


ATOM
351
O
LEU
L
46
−31.747
−12.088
5.179
1
26.72
L


ATOM
352
N
LEU
L
47
−32.994
−10.833
6.542
1
20.56
L


ATOM
353
CA
LEU
L
47
−33.885
−11.94
6.882
1
19.62
L


ATOM
354
CB
LEU
L
47
−33.985
−12.107
8.402
1
15.41
L


ATOM
355
CG
LEU
L
47
−32.715
−12.505
9.152
1
16.42
L


ATOM
356
CD1
LEU
L
47
−33.008
−12.532
10.644
1
14.57
L


ATOM
357
CD2
LEU
L
47
−32.208
−13.86
8.657
1
8.77
L


ATOM
358
C
LEU
L
47
−35.291
−11.747
6.339
1
19.36
L


ATOM
359
O
LEU
L
47
−35.845
−12.617
5.67
1
20.87
L


ATOM
360
N
ILE
L
48
−35.854
−10.588
6.641
1
20.51
L


ATOM
361
CA
ILE
L
48
−37.208
−10.25
6.25
1
22.66
L


ATOM
362
CB
ILE
L
48
−38.155
−10.304
7.475
1
23.74
L


ATOM
363
CG2
ILE
L
48
−39.56
−9.871
7.068
1
25.37
L


ATOM
364
CG1
ILE
L
48
−38.165
−11.711
8.076
1
23.46
L


ATOM
365
CD1
ILE
L
48
−38.789
−12.751
7.182
1
21.29
L


ATOM
366
C
ILE
L
48
−37.28
−8.835
5.706
1
22.85
L


ATOM
367
O
ILE
L
48
−36.61
−7.936
6.213
1
23.92
L


ATOM
368
N
SER
L
49
−38.095
−8.644
4.676
1
22.66
L


ATOM
369
CA
SER
L
49
−38.308
−7.32
4.104
1
23.07
L


ATOM
370
CB
SER
L
49
−37.718
−7.212
2.687
1
24.44
L


ATOM
371
OG
SER
L
49
−38.398
−8.042
1.763
1
24.39
L


ATOM
372
C
SER
L
49
−39.825
−7.128
4.069
1
21.99
L


ATOM
373
O
SER
L
49
−40.581
−8.092
3.93
1
20.34
L


ATOM
374
N
TRP
L
50
−40.27
−5.89
4.218
1
22.89
L


ATOM
375
CA
TRP
L
50
−41.699
−5.597
4.203
1
23.76
L


ATOM
376
CB
TRP
L
50
−42.278
−5.815
2.805
1
22.11
L


ATOM
377
CG
TRP
L
50
−41.931
−4.711
1.859
1
21.52
L


ATOM
378
CD2
TRP
L
50
−42.594
−3.441
1.746
1
18.16
L


ATOM
379
CE2
TRP
L
50
−41.928
−2.712
0.732
1
15.73
L


ATOM
380
CE3
TRP
L
50
−43.685
−2.851
2.403
1
13.3
L


ATOM
381
CD1
TRP
L
50
−40.922
−4.7
0.942
1
19.62
L


ATOM
382
NE1
TRP
L
50
−40.914
−3.503
0.262
1
19.88
L


ATOM
383
CZ2
TRP
L
50
−42.317
−1.418
0.354
1
13.28
L


ATOM
384
CZ3
TRP
L
50
−44.073
−1.56
2.028
1
13.71
L


ATOM
385
CH2
TRP
L
50
−43.387
−0.861
1.011
1
12.41
L


ATOM
386
C
TRP
L
50
−42.486
−6.416
5.213
1
20.91
L


ATOM
387
O
TRP
L
50
−43.497
−7.02
4.887
1
22.48
L


ATOM
388
N
ALA
L
51
−41.992
−6.433
6.441
1
21.79
L


ATOM
389
CA
ALA
L
51
−42.635
−7.129
7.543
1
24.98
L


ATOM
390
CB
ALA
L
51
−44.048
−6.565
7.751
1
23.42
L


ATOM
391
C
ALA
L
51
−42.704
−8.648
7.471
1
26.66
L


ATOM
392
O
ALA
L
51
−42.403
−9.317
8.456
1
24.85
L


ATOM
393
N
SER
L
52
−43.085
−9.197
6.321
1
26.55
L


ATOM
394
CA
SER
L
52
−43.24
−10.64
6.223
1
29.15
L


ATOM
395
CB
SER
L
52
−44.735
−10.979
6.17
1
27.5
L


ATOM
396
OG
SER
L
52
−45.347
−10.433
5.008
1
27.5
L


ATOM
397
C
SER
L
52
−42.536
−11.381
5.096
1
32.33
L


ATOM
398
O
SER
L
52
−42.484
−12.613
5.11
1
34.93
L


ATOM
399
N
THR
L
53
−41.999
−10.661
4.119
1
33.9
L


ATOM
400
CA
THR
L
53
−41.336
−11.333
3.008
1
33.22
L


ATOM
401
CB
THR
L
53
−41.073
−10.366
1.853
1
33.67
L


ATOM
402
OG1
THR
L
53
−42.295
−9.703
1.499
1
33.66
L


ATOM
403
CG2
THR
L
53
−40.548
−11.126
0.644
1
33.16
L


ATOM
404
C
THR
L
53
−40.018
−11.965
3.43
1
32.99
L


ATOM
405
O
THR
L
53
−39.082
−11.271
3.811
1
33.34
L


ATOM
406
N
ARG
L
54
−39.951
−13.291
3.364
1
33.79
L


ATOM
407
CA
ARG
L
54
−38.735
−13.999
3.746
1
35.92
L


ATOM
408
CB
ARG
L
54
−39.038
−15.424
4.197
1
32.75
L


ATOM
409
CG
ARG
L
54
−37.793
−16.192
4.587
1
31.53
L


ATOM
410
CD
ARG
L
54
−38.148
−17.568
5.079
1
33.63
L


ATOM
411
NE
ARG
L
54
−39.309
−17.503
5.953
1
37.86
L


ATOM
412
CZ
ARG
L
54
−40.467
−18.095
5.687
1
40.65
L


ATOM
413
NH1
ARG
L
54
−40.607
−18.8
4.574
1
42.43
L


ATOM
414
NH2
ARG
L
54
−41.49
−17.969
6.523
1
43.56
L


ATOM
415
C
ARG
L
54
−37.737
−14.059
2.608
1
37.11
L


ATOM
416
O
ARG
L
54
−38.039
−14.559
1.528
1
38.23
L


ATOM
417
N
HIS
L
55
−36.539
−13.558
2.867
1
38.89
L


ATOM
418
CA
HIS
L
55
−35.491
−13.554
1.87
1
39.76
L


ATOM
419
CB
HIS
L
55
−34.322
−12.715
2.366
1
44.09
L


ATOM
420
CG
HIS
L
55
−33.465
−12.176
1.27
1
49.71
L


ATOM
421
CD2
HIS
L
55
−33.733
−11.95
−0.039
1
52.46
L


ATOM
422
ND1
HIS
L
55
−32.158
−11.788
1.469
1
52.09
L


ATOM
423
CE1
HIS
L
55
−31.655
−11.346
0.329
1
54.31
L


ATOM
424
NE2
HIS
L
55
−32.59
−11.434
−0.601
1
54.67
L


ATOM
425
C
HIS
L
55
−35.033
−14.988
1.61
1
39.61
L


ATOM
426
O
HIS
L
55
−35.236
−15.871
2.442
1
39.28
L


ATOM
427
N
THR
L
56
−34.423
−15.219
0.451
1
41.6
L


ATOM
428
CA
THR
L
56
−33.934
−16.55
0.098
1
42.65
L


ATOM
429
CB
THR
L
56
−33.583
−16.654
−1.401
1
43.79
L


ATOM
430
OG1
THR
L
56
−32.671
−15.607
−1.752
1
45.15
L


ATOM
431
CG2
THR
L
56
−34.836
−16.548
−2.257
1
43.55
L


ATOM
432
C
THR
L
56
−32.684
−16.867
0.905
1
42.41
L


ATOM
433
O
THR
L
56
−31.952
−15.964
1.308
1
43.65
L


ATOM
434
N
GLY
L
57
−32.434
−18.151
1.133
1
41.88
L


ATOM
435
CA
GLY
L
57
−31.274
−18.53
1.912
1
40.66
L


ATOM
436
C
GLY
L
57
−31.583
−18.259
3.369
1
40.72
L


ATOM
437
O
GLY
L
57
−30.679
−18.096
4.196
1
43.22
L


ATOM
438
N
VAL
L
58
−32.879
−18.195
3.668
1
37.17
L


ATOM
439
CA
VAL
L
58
−33.373
−17.956
5.019
1
34.34
L


ATOM
440
CB
VAL
L
58
−34.042
−16.562
5.132
1
32.13
L


ATOM
441
CG1
VAL
L
58
−34.629
−16.363
6.519
1
30.74
L


ATOM
442
CG2
VAL
L
58
−33.025
−15.481
4.853
1
28.46
L


ATOM
443
C
VAL
L
58
−34.408
−19.039
5.315
1
34.51
L


ATOM
444
O
VAL
L
58
−35.437
−19.121
4.643
1
33.67
L


ATOM
445
N
PRO
L
59
−34.142
−19.891
6.319
1
34.54
L


ATOM
446
CD
PRO
L
59
−32.978
−19.84
7.221
1
35.91
L


ATOM
447
CA
PRO
L
59
−35.051
−20.979
6.703
1
35.62
L


ATOM
448
CB
PRO
L
59
−34.287
−21.693
7.819
1
35.06
L


ATOM
449
CG
PRO
L
59
−33.474
−20.611
8.423
1
35.87
L


ATOM
450
C
PRO
L
59
−36.434
−20.503
7.146
1
37.99
L


ATOM
451
O
PRO
L
59
−36.563
−19.528
7.89
1
38.44
L


ATOM
452
N
ASP
L
60
−37.467
−21.209
6.695
1
38.64
L


ATOM
453
CA
ASP
L
60
−38.835
−20.841
7.029
1
40.72
L


ATOM
454
CB
ASP
L
60
−39.833
−21.755
6.313
1
46.03
L


ATOM
455
CG
ASP
L
60
−39.453
−23.218
6.391
1
50.98
L


ATOM
456
OD1
ASP
L
60
−39.135
−23.703
7.507
1
53.57
L


ATOM
457
OD2
ASP
L
60
−39.484
−23.88
5.326
1
53.05
L


ATOM
458
C
ASP
L
60
−39.165
−20.805
8.509
1
38.96
L


ATOM
459
O
ASP
L
60
−40.285
−20.443
8.876
1
39.35
L


ATOM
460
N
ARG
L
61
−38.22
−21.179
9.366
1
36.43
L


ATOM
461
CA
ARG
L
61
−38.495
−21.126
10.797
1
34.89
L


ATOM
462
CB
ARG
L
61
−37.461
−21.917
11.606
1
32.74
L


ATOM
463
CG
ARG
L
61
−36.073
−21.325
11.651
1
31.93
L


ATOM
464
CD
ARG
L
61
−35.364
−21.796
12.909
1
33.12
L


ATOM
465
NE
ARG
L
61
−33.928
−21.579
12.843
1
32.95
L


ATOM
466
CZ
ARG
L
61
−33.155
−22.096
11.896
1
36.68
L


ATOM
467
NH1
ARG
L
61
−33.696
−22.852
10.949
1
40.54
L


ATOM
468
NH2
ARG
L
61
−31.848
−21.863
11.891
1
38.42
L


ATOM
469
C
ARG
L
61
−38.476
−19.655
11.195
1
34.36
L


ATOM
470
O
ARG
L
61
−38.859
−19.291
12.312
1
35.8
L


ATOM
471
N
PHE
L
62
−38.023
−18.817
10.264
1
32.14
L


ATOM
472
CA
PHE
L
62
−37.975
−17.374
10.475
1
32.9
L


ATOM
473
CB
PHE
L
62
−36.7
−16.745
9.888
1
32.92
L


ATOM
474
CG
PHE
L
62
−35.444
−17.057
10.648
1
36.82
L


ATOM
475
CD1
PHE
L
62
−34.86
−18.317
10.575
1
37.85
L


ATOM
476
CD2
PHE
L
62
−34.833
−16.081
11.428
1
37.67
L


ATOM
477
CE1
PHE
L
62
−33.685
−18.597
11.27
1
38.25
L


ATOM
478
CE2
PHE
L
62
−33.66
−16.351
12.126
1
36.8
L


ATOM
479
CZ
PHE
L
62
−33.085
−17.611
12.046
1
38.55
L


ATOM
480
C
PHE
L
62
−39.177
−16.723
9.781
1
32.54
L


ATOM
481
O
PHE
L
62
−39.28
−16.723
8.55
1
29.09
L


ATOM
482
N
THR
L
63
−40.09
−16.178
10.57
1
30.13
L


ATOM
483
CA
THR
L
63
−41.233
−15.503
9.993
1
32.36
L


ATOM
484
CB
THR
L
63
−42.554
−16.272
10.202
1
32.47
L


ATOM
485
OG1
THR
L
63
−42.751
−16.513
11.6
1
30.8
L


ATOM
486
CG2
THR
L
63
−42.536
−17.584
9.425
1
30.63
L


ATOM
487
C
THR
L
63
−41.368
−14.151
10.652
1
34.24
L


ATOM
488
O
THR
L
63
−41.119
−13.993
11.853
1
35.43
L


ATOM
489
N
GLY
L
64
−41.744
−13.17
9.848
1
33.55
L


ATOM
490
CA
GLY
L
64
−41.937
−11.835
10.364
1
31.62
L


ATOM
491
C
GLY
L
64
−43.395
−11.51
10.15
1
27.73
L


ATOM
492
O
GLY
L
64
−44.073
−12.167
9.364
1
26.61
L


ATOM
493
N
SER
L
65
−43.889
−10.515
10.865
1
25.86
L


ATOM
494
CA
SER
L
65
−45.275
−10.118
10.719
1
26.08
L


ATOM
495
CB
SER
L
65
−46.197
−11.079
11.474
1
24.82
L


ATOM
496
OG
SER
L
65
−46.05
−10.925
12.877
1
28.94
L


ATOM
497
C
SER
L
65
−45.362
−8.736
11.318
1
25.33
L


ATOM
498
O
SER
L
65
−44.575
−8.391
12.198
1
26.68
L


ATOM
499
N
GLY
L
66
−46.299
−7.934
10.834
1
23.56
L


ATOM
500
CA
GLY
L
66
−46.425
−6.601
11.377
1
22.21
L


ATOM
501
C
GLY
L
66
−47.062
−5.619
10.431
1
20.55
L


ATOM
502
O
GLY
L
66
−47.261
−5.902
9.251
1
22.08
L


ATOM
503
N
SER
L
67
−47.376
−4.448
10.961
1
20.16
L


ATOM
504
CA
SER
L
67
−48.007
−3.411
10.176
1
22.22
L


ATOM
505
CB
SER
L
67
−49.395
−3.866
9.727
1
21.86
L


ATOM
506
OG
SER
L
67
−49.981
−2.938
8.834
1
24
L


ATOM
507
C
SER
L
67
−48.132
−2.178
11.042
1
23.38
L


ATOM
508
O
SER
L
67
−47.716
−2.171
12.198
1
24.48
L


ATOM
509
N
GLY
L
68
−48.717
−1.134
10.475
1
25.95
L


ATOM
510
CA
GLY
L
68
−48.883
0.091
11.217
1
24.39
L


ATOM
511
C
GLY
L
68
−47.614
0.492
11.931
1
23.89
L


ATOM
512
O
GLY
L
68
−46.69
1.013
11.313
1
25.46
L


ATOM
513
N
THR
L
69
−47.558
0.208
13.227
1
23.66
L


ATOM
514
CA
THR
L
69
−46.426
0.591
14.051
1
24.04
L


ATOM
515
CB
THR
L
69
−46.909
1.585
15.112
1
26.03
L


ATOM
516
OG1
THR
L
69
−45.794
2.302
15.651
1
34.3
L


ATOM
517
CG2
THR
L
69
−47.629
0.843
16.238
1
24.28
L


ATOM
518
C
THR
L
69
−45.695
−0.565
14.754
1
25.45
L


ATOM
519
O
THR
L
69
−44.611
−0.372
15.309
1
23.36
L


ATOM
520
N
ASP
L
70
−46.281
−1.76
14.729
1
25.24
L


ATOM
521
CA
ASP
L
70
−45.686
−2.915
15.396
1
26.45
L


ATOM
522
CB
ASP
L
70
−46.654
−3.473
16.439
1
29.94
L


ATOM
523
CG
ASP
L
70
−46.812
−2.565
17.634
1
32.11
L


ATOM
524
OD1
ASP
L
70
−46.437
−1.377
17.535
1
33.82
L


ATOM
525
OD2
ASP
L
70
−47.323
−3.042
18.673
1
34.38
L


ATOM
526
C
ASP
L
70
−45.295
−4.034
14.447
1
27.14
L


ATOM
527
O
ASP
L
70
−46.069
−4.424
13.574
1
27.48
L


ATOM
528
N
TYR
L
71
−44.091
−4.562
14.648
1
27.09
L


ATOM
529
CA
TYR
L
71
−43.56
−5.636
13.82
1
25.67
L


ATOM
530
CB
TYR
L
71
−42.584
−5.041
12.789
1
22.93
L


ATOM
531
CG
TYR
L
71
−43.252
−3.965
11.963
1
19.63
L


ATOM
532
CD1
TYR
L
71
−43.514
−2.701
12.505
1
16.75
L


ATOM
533
CE1
TYR
L
71
−44.28
−1.758
11.812
1
16.18
L


ATOM
534
CD2
TYR
L
71
−43.758
−4.253
10.696
1
19.84
L


ATOM
535
CE2
TYR
L
71
−44.521
−3.319
9.994
1
22.27
L


ATOM
536
CZ
TYR
L
71
−44.783
−2.075
10.562
1
20.24
L


ATOM
537
OH
TYR
L
71
−45.583
−1.18
9.884
1
22.36
L


ATOM
538
C
TYR
L
71
−42.882
−6.704
14.68
1
26.55
L


ATOM
539
O
TYR
L
71
−42.192
−6.398
15.661
1
26.09
L


ATOM
540
N
THR
L
72
−43.094
−7.961
14.313
1
26.01
L


ATOM
541
CA
THR
L
72
−42.518
−9.067
15.056
1
27.48
L


ATOM
542
CB
THR
L
72
−43.602
−9.87
15.804
1
27.52
L


ATOM
543
OG1
THR
L
72
−44.351
−8.992
16.648
1
30.44
L


ATOM
544
CG2
THR
L
72
−42.962
−10.957
16.666
1
28.87
L


ATOM
545
C
THR
L
72
−41.752
−10.035
14.169
1
28.75
L


ATOM
546
O
THR
L
72
−42.17
−10.358
13.052
1
29.88
L


ATOM
547
N
LEU
L
73
−40.616
−10.483
14.685
1
28.5
L


ATOM
548
CA
LEU
L
73
−39.772
−11.443
13.999
1
30.08
L


ATOM
549
CB
LEU
L
73
−38.326
−10.93
13.925
1
30.95
L


ATOM
550
CG
LEU
L
73
−37.209
−11.719
13.22
1
32.74
L


ATOM
551
CD1
LEU
L
73
−36.737
−12.849
14.102
1
38.04
L


ATOM
552
CD2
LEU
L
73
−37.697
−12.254
11.886
1
32.53
L


ATOM
553
C
LEU
L
73
−39.871
−12.675
14.884
1
30.34
L


ATOM
554
O
LEU
L
73
−39.398
−12.672
16.019
1
31.95
L


ATOM
555
N
THR
L
74
−40.532
−13.711
14.384
1
28.68
L


ATOM
556
CA
THR
L
74
−40.669
−14.931
15.155
1
26.67
L


ATOM
557
CB
THR
L
74
−42.118
−15.483
15.124
1
27.43
L


ATOM
558
OG1
THR
L
74
−43.005
−14.586
15.81
1
24.66
L


ATOM
559
CG2
THR
L
74
−42.17
−16.854
15.793
1
26
L


ATOM
560
C
THR
L
74
−39.731
−15.999
14.608
1
25.39
L


ATOM
561
O
THR
L
74
−39.604
−16.177
13.396
1
21.17
L


ATOM
562
N
ILE
L
75
−39.051
−16.684
15.518
1
26.91
L


ATOM
563
CA
ILE
L
75
−38.154
−17.761
15.137
1
27.42
L


ATOM
564
CB
ILE
L
75
−36.715
−17.538
15.667
1
23.84
L


ATOM
565
CG2
ILE
L
75
−35.8
−18.7
15.225
1
24.53
L


ATOM
566
CG1
ILE
L
75
−36.164
−16.209
15.145
1
19.54
L


ATOM
567
CD1
ILE
L
75
−34.723
−15.917
15.577
1
18.05
L


ATOM
568
C
ILE
L
75
−38.731
−19.025
15.759
1
29.37
L


ATOM
569
O
ILE
L
75
−38.614
−19.244
16.963
1
27.99
L


ATOM
570
N
SER
L
76
−39.39
−19.837
14.94
1
32.4
L


ATOM
571
CA
SER
L
76
−39.972
−21.085
15.422
1
35.69
L


ATOM
572
CB
SER
L
76
−40.994
−21.619
14.425
1
34.27
L


ATOM
573
OG
SER
L
76
−42.058
−20.705
14.251
1
41.41
L


ATOM
574
C
SER
L
76
−38.854
−22.102
15.58
1
37.52
L


ATOM
575
O
SER
L
76
−37.901
−22.108
14.795
1
39.37
L


ATOM
576
N
SER
L
77
−38.963
−22.956
16.59
1
37.95
L


ATOM
577
CA
SER
L
77
−37.944
−23.971
16.817
1
39.62
L


ATOM
578
CB
SER
L
77
−38.088
−25.087
15.778
1
39.48
L


ATOM
579
OG
SER
L
77
−36.932
−25.903
15.738
1
43.84
L


ATOM
580
C
SER
L
77
−36.547
−23.36
16.732
1
38.3
L


ATOM
581
O
SER
L
77
−35.786
−23.654
15.814
1
41.65
L


ATOM
582
N
VAL
L
78
−36.217
−22.504
17.69
1
37.08
L


ATOM
583
CA
VAL
L
78
−34.914
−21.854
17.714
1
33.69
L


ATOM
584
CB
VAL
L
78
−34.782
−20.971
18.955
1
32.5
L


ATOM
585
CG1
VAL
L
78
−33.444
−20.258
18.951
1
32.68
L


ATOM
586
CG2
VAL
L
78
−35.917
−19.967
18.984
1
34.67
L


ATOM
587
C
VAL
L
78
−33.771
−22.865
17.7
1
34.31
L


ATOM
588
O
VAL
L
78
−33.877
−23.947
18.277
1
35.13
L


ATOM
589
N
GLN
L
79
−32.681
−22.506
17.027
1
34.75
L


ATOM
590
CA
GLN
L
79
−31.503
−23.361
16.933
1
33.83
L


ATOM
591
CB
GLN
L
79
−31.245
−23.758
15.48
1
35.05
L


ATOM
592
CG
GLN
L
79
−31.467
−25.229
15.195
1
40.46
L


ATOM
593
CD
GLN
L
79
−32.895
−25.669
15.454
1
44.87
L


ATOM
594
OE1
GLN
L
79
−33.814
−25.307
14.717
1
44.9
L


ATOM
595
NE2
GLN
L
79
−33.091
−26.456
16.513
1
46.97
L


ATOM
596
C
GLN
L
79
−30.283
−22.64
17.486
1
33.42
L


ATOM
597
O
GLN
L
79
−30.297
−21.424
17.68
1
32.99
L


ATOM
598
N
ALA
L
80
−29.221
−23.393
17.739
1
33.27
L


ATOM
599
CA
ALA
L
80
−28.003
−22.807
18.272
1
31.75
L


ATOM
600
CB
ALA
L
80
−27.053
−23.904
18.703
1
32.15
L


ATOM
601
C
ALA
L
80
−27.336
−21.896
17.243
1
32.69
L


ATOM
602
O
ALA
L
80
−26.637
−20.947
17.604
1
31.8
L


ATOM
603
N
GLU
L
81
−27.558
−22.182
15.963
1
32.33
L


ATOM
604
CA
GLU
L
81
−26.974
−21.382
14.892
1
32.85
L


ATOM
605
CB
GLU
L
81
−27.006
−22.153
13.566
1
36.21
L


ATOM
606
CG
GLU
L
81
−28.411
−22.437
13.046
1
44.3
L


ATOM
607
CD
GLU
L
81
−28.438
−23.334
11.81
1
48.2
L


ATOM
608
OE1
GLU
L
81
−27.753
−23.01
10.81
1
50.13
L


ATOM
609
OE2
GLU
L
81
−29.158
−24.36
11.839
1
49.51
L


ATOM
610
C
GLU
L
81
−27.73
−20.069
14.741
1
31.6
L


ATOM
611
O
GLU
L
81
−27.373
−19.233
13.914
1
32.12
L


ATOM
612
N
ASP
L
82
−28.78
−19.889
15.536
1
29.09
L


ATOM
613
CA
ASP
L
82
−29.555
−18.659
15.47
1
29.13
L


ATOM
614
CB
ASP
L
82
−31.044
−18.942
15.688
1
28.38
L


ATOM
615
CG
ASP
L
82
−31.611
−19.894
14.648
1
31.59
L


ATOM
616
OD1
ASP
L
82
−31.344
−19.691
13.439
1
30.2
L


ATOM
617
OD2
ASP
L
82
−32.329
−20.843
15.034
1
30.04
L


ATOM
618
C
ASP
L
82
−29.054
−17.636
16.481
1
28.07
L


ATOM
619
O
ASP
L
82
−29.578
−16.525
16.562
1
26.56
L


ATOM
620
N
LEU
L
83
−28.044
−18.014
17.26
1
27.17
L


ATOM
621
CA
LEU
L
83
−27.465
−17.086
18.226
1
26.08
L


ATOM
622
CB
LEU
L
83
−26.355
−17.755
19.053
1
25.04
L


ATOM
623
CG
LEU
L
83
−26.718
−18.882
20.031
1
24.92
L


ATOM
624
CD1
LEU
L
83
−25.444
−19.443
20.667
1
20.63
L


ATOM
625
CD2
LEU
L
83
−27.666
−18.361
21.094
1
20.79
L


ATOM
626
C
LEU
L
83
−26.87
−15.954
17.393
1
25.94
L


ATOM
627
O
LEU
L
83
−26.023
−16.184
16.526
1
24.91
L


ATOM
628
N
ALA
L
84
−27.325
−14.735
17.648
1
25.72
L


ATOM
629
CA
ALA
L
84
−26.845
−13.586
16.903
1
25.28
L


ATOM
630
CB
ALA
L
84
−27.113
−13.796
15.417
1
24.65
L


ATOM
631
C
ALA
L
84
−27.52
−12.3
17.374
1
26.49
L


ATOM
632
O
ALA
L
84
−28.344
−12.303
18.297
1
23.91
L


ATOM
633
N
LEU
L
85
−27.149
−11.198
16.733
1
24.66
L


ATOM
634
CA
LEU
L
85
−27.718
−9.902
17.042
1
23
L


ATOM
635
CB
LEU
L
85
−26.618
−8.846
17.01
1
22.46
L


ATOM
636
CG
LEU
L
85
−26.858
−7.554
17.791
1
26.05
L


ATOM
637
CD1
LEU
L
85
−27.387
−7.838
19.199
1
23.12
L


ATOM
638
CD2
LEU
L
85
−25.543
−6.8
17.854
1
26.91
L


ATOM
639
C
LEU
L
85
−28.761
−9.657
15.948
1
23.66
L


ATOM
640
O
LEU
L
85
−28.496
−9.895
14.769
1
22.22
L


ATOM
641
N
TYR
L
86
−29.954
−9.217
16.341
1
23.7
L


ATOM
642
CA
TYR
L
86
−31.028
−8.977
15.385
1
22.23
L


ATOM
643
CB
TYR
L
86
−32.208
−9.912
15.669
1
20.35
L


ATOM
644
CG
TYR
L
86
−31.893
−11.382
15.457
1
21.38
L


ATOM
645
CD1
TYR
L
86
−31.326
−12.154
16.471
1
20.46
L


ATOM
646
CE1
TYR
L
86
−30.983
−13.49
16.259
1
16.21
L


ATOM
647
CD2
TYR
L
86
−32.114
−11.987
14.22
1
21.25
L


ATOM
648
CE2
TYR
L
86
−31.772
−13.317
13.995
1
21.34
L


ATOM
649
CZ
TYR
L
86
−31.205
−14.061
15.019
1
19.18
L


ATOM
650
OH
TYR
L
86
−30.839
−15.365
14.781
1
20.03
L


ATOM
651
C
TYR
L
86
−31.511
−7.532
15.385
1
24.29
L


ATOM
652
O
TYR
L
86
−31.887
−6.983
16.428
1
24.91
L


ATOM
653
N
TYR
L
87
−31.489
−6.911
14.21
1
21.91
L


ATOM
654
CA
TYR
L
87
−31.937
−5.537
14.093
1
22.37
L


ATOM
655
CB
TYR
L
87
−30.843
−4.616
13.553
1
23.27
L


ATOM
656
CG
TYR
L
87
−29.551
−4.574
14.319
1
28.68
L


ATOM
657
CD1
TYR
L
87
−28.578
−5.553
14.129
1
28.71
L


ATOM
658
CE1
TYR
L
87
−27.352
−5.479
14.779
1
33.56
L


ATOM
659
CD2
TYR
L
87
−29.269
−3.518
15.188
1
29.31
L


ATOM
660
CE2
TYR
L
87
−28.043
−3.432
15.843
1
31.52
L


ATOM
661
CZ
TYR
L
87
−27.086
−4.415
15.634
1
33.34
L


ATOM
662
OH
TYR
L
87
−25.855
−4.336
16.26
1
35.51
L


ATOM
663
C
TYR
L
87
−33.096
−5.434
13.126
1
24
L


ATOM
664
O
TYR
L
87
−33.215
−6.214
12.178
1
20.82
L


ATOM
665
N
CYS
L
88
−33.954
−4.458
13.379
1
23.57
L


ATOM
666
CA
CYS
L
88
−35.062
−4.194
12.496
1
25.15
L


ATOM
667
C
CYS
L
88
−34.712
−2.841
11.912
1
23.75
L


ATOM
668
O
CYS
L
88
−33.978
−2.062
12.521
1
23.73
L


ATOM
669
CB
CYS
L
88
−36.389
−4.149
13.258
1
26.56
L


ATOM
670
SG
CYS
L
88
−36.559
−2.976
14.646
1
31.43
L


ATOM
671
N
GLN
L
89
−35.219
−2.565
10.724
1
24.24
L


ATOM
672
CA
GLN
L
89
−34.931
−1.307
10.062
1
22.23
L


ATOM
673
CB
GLN
L
89
−33.747
−1.492
9.111
1
19.89
L


ATOM
674
CG
GLN
L
89
−33.406
−0.283
8.27
1
21.51
L


ATOM
675
CD
GLN
L
89
−33.483
−0.584
6.782
1
21.16
L


ATOM
676
OE1
GLN
L
89
−32.872
−1.539
6.3
1
22.19
L


ATOM
677
NE2
GLN
L
89
−34.231
0.231
6.049
1
17.88
L


ATOM
678
C
GLN
L
89
−36.161
−0.907
9.287
1
21.86
L


ATOM
679
O
GLN
L
89
−36.757
−1.741
8.604
1
23.52
L


ATOM
680
N
GLN
L
90
−36.566
0.352
9.396
1
19.94
L


ATOM
681
CA
GLN
L
90
−37.732
0.773
8.641
1
20.67
L


ATOM
682
CB
GLN
L
90
−38.545
1.836
9.406
1
19.9
L


ATOM
683
CG
GLN
L
90
−37.805
3.097
9.807
1
20.58
L


ATOM
684
CD
GLN
L
90
−37.73
4.137
8.698
1
24.36
L


ATOM
685
OE1
GLN
L
90
−38.593
4.189
7.808
1
19.07
L


ATOM
686
NE2
GLN
L
90
−36.702
4.994
8.762
1
20.34
L


ATOM
687
C
GLN
L
90
−37.268
1.305
7.304
1
19.46
L


ATOM
688
O
GLN
L
90
−36.249
1.991
7.223
1
16.88
L


ATOM
689
N
HIS
L
91
−37.984
0.939
6.248
1
18.39
L


ATOM
690
CA
HIS
L
91
−37.648
1.428
4.921
1
22.42
L


ATOM
691
CB
HIS
L
91
−37.153
0.285
3.993
1
23.41
L


ATOM
692
CG
HIS
L
91
−37.957
−0.985
4.048
1
24.78
L


ATOM
693
CD2
HIS
L
91
−37.791
−2.111
4.788
1
20.04
L


ATOM
694
ND1
HIS
L
91
−39.017
−1.24
3.2
1
23.86
L


ATOM
695
CE1
HIS
L
91
−39.463
−2.468
3.411
1
21.59
L


ATOM
696
NE2
HIS
L
91
−38.736
−3.017
4.369
1
17.36
L


ATOM
697
C
HIS
L
91
−38.842
2.186
4.341
1
21.15
L


ATOM
698
O
HIS
L
91
−39.143
2.117
3.151
1
21.21
L


ATOM
699
N
TYR
L
92
−39.489
2.944
5.225
1
19.56
L


ATOM
700
CA
TYR
L
92
−40.666
3.748
4.908
1
19.53
L


ATOM
701
CB
TYR
L
92
−41.53
3.905
6.168
1
21.02
L


ATOM
702
CG
TYR
L
92
−42.787
4.731
6.001
1
18.63
L


ATOM
703
CD1
TYR
L
92
−43.81
4.316
5.138
1
21.58
L


ATOM
704
CE1
TYR
L
92
−44.983
5.062
4.989
1
21.19
L


ATOM
705
CD2
TYR
L
92
−42.965
5.92
6.714
1
18.34
L


ATOM
706
CE2
TYR
L
92
−44.137
6.681
6.575
1
19.57
L


ATOM
707
CZ
TYR
L
92
−45.136
6.242
5.708
1
23.05
L


ATOM
708
OH
TYR
L
92
−46.273
6.985
5.531
1
24.67
L


ATOM
709
C
TYR
L
92
−40.27
5.124
4.396
1
19.03
L


ATOM
710
O
TYR
L
92
−40.909
5.681
3.503
1
18.47
L


ATOM
711
N
THR
L
93
−39.214
5.68
4.967
1
19.88
L


ATOM
712
CA
THR
L
93
−38.775
6.996
4.541
1
20.71
L


ATOM
713
CB
THR
L
93
−39.764
8.069
5.016
1
20.02
L


ATOM
714
OG1
THR
L
93
−39.56
9.272
4.262
1
22.94
L


ATOM
715
CG2
THR
L
93
−39.574
8.344
6.507
1
12.63
L


ATOM
716
C
THR
L
93
−37.389
7.331
5.074
1
22.15
L


ATOM
717
O
THR
L
93
−36.919
6.727
6.038
1
25
L


ATOM
718
N
THR
L
94
−36.73
8.291
4.44
1
21.44
L


ATOM
719
CA
THR
L
94
−35.408
8.68
4.889
1
23.37
L


ATOM
720
CB
THR
L
94
−34.56
9.259
3.731
1
21.39
L


ATOM
721
OG1
THR
L
94
−35.141
10.479
3.267
1
21.96
L


ATOM
722
CG2
THR
L
94
−34.492
8.267
2.584
1
21.61
L


ATOM
723
C
THR
L
94
−35.566
9.724
5.99
1
22.48
L


ATOM
724
O
THR
L
94
−36.519
10.5
5.988
1
24.55
L


ATOM
725
N
PRO
L
95
−34.64
9.741
6.953
1
20.82
L


ATOM
726
CD
PRO
L
95
−34.57
10.684
8.084
1
19.76
L


ATOM
727
CA
PRO
L
95
−33.509
8.813
6.98
1
22.19
L


ATOM
728
CB
PRO
L
95
−32.566
9.466
7.984
1
20.89
L


ATOM
729
CG
PRO
L
95
−33.531
10.035
8.981
1
22.02
L


ATOM
730
C
PRO
L
95
−33.91
7.405
7.412
1
21.88
L


ATOM
731
O
PRO
L
95
−34.825
7.23
8.215
1
25.53
L


ATOM
732
N
LEU
L
96
−33.223
6.406
6.874
1
19.75
L


ATOM
733
CA
LEU
L
96
−33.485
5.025
7.242
1
19.84
L


ATOM
734
CB
LEU
L
96
−32.757
4.074
6.291
1
17.28
L


ATOM
735
CG
LEU
L
96
−33.179
4.145
4.826
1
15.85
L


ATOM
736
CD1
LEU
L
96
−32.438
3.078
4.015
1
11.63
L


ATOM
737
CD2
LEU
L
96
−34.688
3.951
4.739
1
15.01
L


ATOM
738
C
LEU
L
96
−32.969
4.821
8.665
1
21.55
L


ATOM
739
O
LEU
L
96
−31.857
5.244
8.992
1
22.96
L


ATOM
740
N
THR
L
97
−33.764
4.164
9.504
1
22.43
L


ATOM
741
CA
THR
L
97
−33.372
3.94
10.895
1
21.97
L


ATOM
742
CB
THR
L
97
−34.191
4.836
11.815
1
18.78
L


ATOM
743
OG1
THR
L
97
−35.572
4.781
11.43
1
17.77
L


ATOM
744
CG2
THR
L
97
−33.695
6.262
11.718
1
16.65
L


ATOM
745
C
THR
L
97
−33.47
2.496
11.39
1
22.79
L


ATOM
746
O
THR
L
97
−34.374
1.754
11.016
1
25.6
L


ATOM
747
N
PHE
L
98
−32.528
2.109
12.243
1
24.33
L


ATOM
748
CA
PHE
L
98
−32.492
0.754
12.786
1
25.34
L


ATOM
749
CB
PHE
L
98
−31.098
0.136
12.614
1
24.41
L


ATOM
750
CG
PHE
L
98
−30.609
0.111
11.207
1
24.08
L


ATOM
751
CD1
PHE
L
98
−30.178
1.276
10.585
1
23.59
L


ATOM
752
CD2
PHE
L
98
−30.58
−1.084
10.493
1
24.56
L


ATOM
753
CE1
PHE
L
98
−29.726
1.248
9.27
1
23.14
L


ATOM
754
CE2
PHE
L
98
−30.129
−1.119
9.175
1
23.11
L


ATOM
755
CZ
PHE
L
98
−29.703
0.046
8.565
1
19.74
L


ATOM
756
C
PHE
L
98
−32.819
0.701
14.267
1
27.37
L


ATOM
757
O
PHE
L
98
−32.667
1.689
14.988
1
26.55
L


ATOM
758
N
GLY
L
99
−33.262
−0.467
14.718
1
28.26
L


ATOM
759
CA
GLY
L
99
−33.537
−0.643
16.128
1
29.27
L


ATOM
760
C
GLY
L
99
−32.179
−0.92
16.76
1
29.59
L


ATOM
761
O
GLY
L
99
−31.223
−1.247
16.051
1
29.47
L


ATOM
762
N
ALA
L
100
−32.084
−0.801
18.08
1
29.37
L


ATOM
763
CA
ALA
L
100
−30.824
−1.029
18.778
1
29.08
L


ATOM
764
CB
ALA
L
100
−30.967
−0.623
20.241
1
29.66
L


ATOM
765
C
ALA
L
100
−30.347
−2.475
18.677
1
29.38
L


ATOM
766
O
ALA
L
100
−29.187
−2.779
18.954
1
30.49
L


ATOM
767
N
GLY
L
101
−31.244
−3.367
18.284
1
29.12
L


ATOM
768
CA
GLY
L
101
−30.871
−4.762
18.154
1
29.81
L


ATOM
769
C
GLY
L
101
−31.219
−5.594
19.375
1
29.76
L


ATOM
770
O
GLY
L
101
−31.258
−5.091
20.497
1
29.97
L


ATOM
771
N
THR
L
102
−31.491
−6.872
19.144
1
29.24
L


ATOM
772
CA
THR
L
102
−31.817
−7.8
20.215
1
29.33
L


ATOM
773
CB
THR
L
102
−33.287
−8.287
20.116
1
28.76
L


ATOM
774
OG1
THR
L
102
−34.174
−7.194
20.398
1
29.56
L


ATOM
775
CG2
THR
L
102
−33.554
−9.403
21.112
1
29.19
L


ATOM
776
C
THR
L
102
−30.86
−8.99
20.117
1
29.39
L


ATOM
777
O
THR
L
102
−30.742
−9.627
19.068
1
27.17
L


ATOM
778
N
LYS
L
103
−30.16
−9.266
21.21
1
29.91
L


ATOM
779
CA
LYS
L
103
−29.212
−10.365
21.243
1
33.61
L


ATOM
780
CB
LYS
L
103
−28.046
−10.015
22.172
1
33.62
L


ATOM
781
CG
LYS
L
103
−26.929
−11.053
22.205
1
36.67
L


ATOM
782
CD
LYS
L
103
−25.893
−10.716
23.282
1
38.03
L


ATOM
783
CE
LYS
L
103
−24.961
−11.895
23.533
1
41.32
L


ATOM
784
NZ
LYS
L
103
−24.031
−11.696
24.697
1
44.17
L


ATOM
785
C
LYS
L
103
−29.904
−11.651
21.709
1
34.54
L


ATOM
786
O
LYS
L
103
−30.346
−11.754
22.854
1
36.77
L


ATOM
787
N
LEU
L
104
−30.011
−12.62
20.805
1
34
L


ATOM
788
CA
LEU
L
104
−30.639
−13.888
21.129
1
32.56
L


ATOM
789
CB
LEU
L
104
−31.074
−14.624
19.861
1
33.44
L


ATOM
790
CG
LEU
L
104
−31.886
−15.91
20.066
1
32.97
L


ATOM
791
CD1
LEU
L
104
−32.632
−16.262
18.783
1
36.14
L


ATOM
792
CD2
LEU
L
104
−30.975
−17.047
20.484
1
32.63
L


ATOM
793
C
LEU
L
104
−29.655
−14.742
21.902
1
32.52
L


ATOM
794
O
LEU
L
104
−28.535
−14.985
21.456
1
27.38
L


ATOM
795
N
GLU
L
105
−30.095
−15.187
23.071
1
33.48
L


ATOM
796
CA
GLU
L
105
−29.289
−16.012
23.946
1
31.91
L


ATOM
797
CB
GLU
L
105
−28.951
−15.226
25.194
1
33.27
L


ATOM
798
CG
GLU
L
105
−28.427
−13.85
24.876
1
35.49
L


ATOM
799
CD
GLU
L
105
−28.069
−13.085
26.118
1
37.33
L


ATOM
800
OE1
GLU
L
105
−28.985
−12.763
26.907
1
36.87
L


ATOM
801
OE2
GLU
L
105
−26.865
−12.818
26.306
1
39.96
L


ATOM
802
C
GLU
L
105
−30.112
−17.231
24.298
1
30.52
L


ATOM
803
O
GLU
L
105
−31.333
−17.19
24.253
1
31.61
L


ATOM
804
N
LEU
L
106
−29.443
−18.316
24.654
1
30.57
L


ATOM
805
CA
LEU
L
106
−30.134
−19.546
24.984
1
29.34
L


ATOM
806
CB
LEU
L
106
−29.666
−20.666
24.057
1
28.9
L


ATOM
807
CG
LEU
L
106
−29.949
−20.475
22.567
1
28.82
L


ATOM
808
CD1
LEU
L
106
−29.112
−21.461
21.758
1
26.11
L


ATOM
809
CD2
LEU
L
106
−31.437
−20.66
22.303
1
26.85
L


ATOM
810
C
LEU
L
106
−29.979
−20.004
26.418
1
30.22
L


ATOM
811
O
LEU
L
106
−28.968
−19.761
27.073
1
29.39
L


ATOM
812
N
LYS
L
107
−31.018
−20.672
26.894
1
33.45
L


ATOM
813
CA
LYS
L
107
−31.042
−21.234
28.226
1
33.81
L


ATOM
814
CB
LYS
L
107
−32.449
−21.141
28.814
1
33.84
L


ATOM
815
CG
LYS
L
107
−32.842
−19.75
29.232
1
37.64
L


ATOM
816
CD
LYS
L
107
−31.984
−19.28
30.4
1
45.3
L


ATOM
817
CE
LYS
L
107
−32.175
−20.179
31.63
1
49.65
L


ATOM
818
NZ
LYS
L
107
−31.369
−19.742
32.81
1
51.56
L


ATOM
819
C
LYS
L
107
−30.672
−22.694
28.031
1
35.03
L


ATOM
820
O
LYS
L
107
−31.213
−23.357
27.143
1
35.03
L


ATOM
821
N
ARG
L
108
−29.737
−23.185
28.838
1
33.52
L


ATOM
822
CA
ARG
L
108
−29.327
−24.579
28.76
1
31.49
L


ATOM
823
CB
ARG
L
108
−28.151
−24.741
27.791
1
31.04
L


ATOM
824
CG
ARG
L
108
−26.917
−23.91
28.111
1
32.01
L


ATOM
825
CD
ARG
L
108
−25.676
−24.664
27.669
1
30.51
L


ATOM
826
NE
ARG
L
108
−25.535
−25.897
28.441
1
34.19
L


ATOM
827
CZ
ARG
L
108
−24.851
−26.966
28.046
1
34.38
L


ATOM
828
NH1
ARG
L
108
−24.233
−26.97
26.872
1
36.37
L


ATOM
829
NH2
ARG
L
108
−24.781
−28.032
28.834
1
35.47
L


ATOM
830
C
ARG
L
108
−28.959
−25.113
30.146
1
31.09
L


ATOM
831
O
ARG
L
108
−29.008
−24.384
31.139
1
32.47
L


ATOM
832
N
ALA
L
109
−28.613
−26.391
30.219
1
29.57
L


ATOM
833
CA
ALA
L
109
−28.247
−26.992
31.495
1
28.76
L


ATOM
834
CB
ALA
L
109
−28.254
−28.516
31.384
1
28.18
L


ATOM
835
C
ALA
L
109
−26.866
−26.506
31.902
1
28.65
L


ATOM
836
O
ALA
L
109
−26.028
−26.21
31.048
1
28.71
L


ATOM
837
N
ASP
L
110
−26.631
−26.418
33.207
1
27.68
L


ATOM
838
CA
ASP
L
110
−25.337
−25.98
33.697
1
26.42
L


ATOM
839
CB
ASP
L
110
−25.295
−26.021
35.219
1
26.12
L


ATOM
840
CG
ASP
L
110
−26.256
−25.055
35.843
1
26.37
L


ATOM
841
OD1
ASP
L
110
−26.691
−24.127
35.129
1
22.25
L


ATOM
842
OD2
ASP
L
110
−26.568
−25.216
37.044
1
29.32
L


ATOM
843
C
ASP
L
110
−24.265
−26.893
33.144
1
25.99
L


ATOM
844
O
ASP
L
110
−24.548
−28.024
32.758
1
25.07
L


ATOM
845
N
ALA
L
111
−23.036
−26.395
33.106
1
24.91
L


ATOM
846
CA
ALA
L
111
−21.914
−27.175
32.609
1
24.41
L


ATOM
847
CB
ALA
L
111
−21.857
−27.115
31.093
1
21.51
L


ATOM
848
C
ALA
L
111
−20.644
−26.605
33.206
1
24.54
L


ATOM
849
O
ALA
L
111
−20.408
−25.398
33.146
1
24.68
L


ATOM
850
N
ALA
L
112
−19.833
−27.482
33.788
1
26.58
L


ATOM
851
CA
ALA
L
112
−18.574
−27.085
34.415
1
28.57
L


ATOM
852
CB
ALA
L
112
−18.067
−28.207
35.325
1
27.25
L


ATOM
853
C
ALA
L
112
−17.51
−26.737
33.379
1
27.17
L


ATOM
854
O
ALA
L
112
−17.285
−27.485
32.43
1
28.07
L


ATOM
855
N
PRO
L
113
−16.83
−25.599
33.561
1
24.92
L


ATOM
856
CD
PRO
L
113
−16.9
−24.689
34.719
1
26.19
L


ATOM
857
CA
PRO
L
113
−15.788
−25.168
32.63
1
24.99
L


ATOM
858
CB
PRO
L
113
−15.409
−23.789
33.155
1
23.49
L


ATOM
859
CG
PRO
L
113
−15.571
−23.953
34.625
1
22.47
L


ATOM
860
C
PRO
L
113
−14.588
−26.092
32.59
1
24.43
L


ATOM
861
O
PRO
L
113
−14.271
−26.761
33.564
1
26.16
L


ATOM
862
N
THR
L
114
−13.932
−26.138
31.443
1
24.86
L


ATOM
863
CA
THR
L
114
−12.733
−26.934
31.301
1
25.05
L


ATOM
864
CB
THR
L
114
−12.706
−27.712
29.972
1
27.63
L


ATOM
865
OG1
THR
L
114
−13.745
−28.702
29.976
1
32.5
L


ATOM
866
CG2
THR
L
114
−11.363
−28.411
29.792
1
28.26
L


ATOM
867
C
THR
L
114
−11.648
−25.869
31.322
1
23.47
L


ATOM
868
O
THR
L
114
−11.55
−25.048
30.411
1
27.13
L


ATOM
869
N
VAL
L
115
−10.86
−25.876
32.387
1
19.01
L


ATOM
870
CA
VAL
L
115
−9.805
−24.905
32.586
1
16.43
L


ATOM
871
CB
VAL
L
115
−9.637
−24.628
34.085
1
15.97
L


ATOM
872
CG1
VAL
L
115
−8.551
−23.585
34.311
1
10.09
L


ATOM
873
CG2
VAL
L
115
−10.983
−24.208
34.681
1
10.52
L


ATOM
874
C
VAL
L
115
−8.458
−25.322
32.014
1
19.89
L


ATOM
875
O
VAL
L
115
−7.977
−26.423
32.265
1
21.12
L


ATOM
876
N
SER
L
116
−7.849
−24.423
31.251
1
21.56
L


ATOM
877
CA
SER
L
116
−6.55
−24.674
30.653
1
23.75
L


ATOM
878
CB
SER
L
116
−6.656
−24.72
29.126
1
24.69
L


ATOM
879
OG
SER
L
116
−7.572
−25.708
28.697
1
27.61
L


ATOM
880
C
SER
L
116
−5.647
−23.519
31.05
1
24.78
L


ATOM
881
O
SER
L
116
−6.069
−22.365
31.026
1
25.92
L


ATOM
882
N
ILE
L
117
−4.406
−23.819
31.413
1
25.28
L


ATOM
883
CA
ILE
L
117
−3.482
−22.759
31.797
1
23.6
L


ATOM
884
CB
ILE
L
117
−3.084
−22.875
33.284
1
23.08
L


ATOM
885
CG2
ILE
L
117
−2.101
−24.022
33.479
1
17.48
L


ATOM
886
CG1
ILE
L
117
−2.522
−21.532
33.762
1
23.72
L


ATOM
887
CD1
ILE
L
117
−2.149
−21.5
35.228
1
22.35
L


ATOM
888
C
ILE
L
117
−2.252
−22.825
30.904
1
20.29
L


ATOM
889
O
ILE
L
117
−1.785
−23.899
30.56
1
20.43
L


ATOM
890
N
PHE
L
118
−1.737
−21.664
30.53
1
20.61
L


ATOM
891
CA
PHE
L
118
−0.598
−21.589
29.633
1
21.15
L


ATOM
892
CB
PHE
L
118
−1.065
−21.101
28.268
1
18.27
L


ATOM
893
CG
PHE
L
118
−2.134
−21.947
27.671
1
19.43
L


ATOM
894
CD1
PHE
L
118
−1.809
−23.074
26.93
1
17.8
L


ATOM
895
CD2
PHE
L
118
−3.472
−21.642
27.884
1
19.5
L


ATOM
896
CE1
PHE
L
118
−2.791
−23.883
26.411
1
18.23
L


ATOM
897
CE2
PHE
L
118
−4.465
−22.444
27.371
1
21.64
L


ATOM
898
CZ
PHE
L
118
−4.128
−23.571
26.628
1
21.36
L


ATOM
899
C
PHE
L
118
0.499
−20.669
30.106
1
21.75
L


ATOM
900
O
PHE
L
118
0.237
−19.567
30.584
1
23.16
L


ATOM
901
N
PRO
L
119
1.753
−21.112
29.975
1
22.12
L


ATOM
902
CD
PRO
L
119
2.198
−22.482
29.669
1
21.31
L


ATOM
903
CA
PRO
L
119
2.885
−20.288
30.391
1
23.47
L


ATOM
904
CB
PRO
L
119
3.998
−21.313
30.587
1
24.07
L


ATOM
905
CG
PRO
L
119
3.701
−22.301
29.525
1
22.51
L


ATOM
906
C
PRO
L
119
3.214
−19.314
29.27
1
23.79
L


ATOM
907
O
PRO
L
119
2.684
−19.421
28.156
1
20.05
L


ATOM
908
N
PRO
L
120
4.087
−18.34
29.553
1
26.14
L


ATOM
909
CD
PRO
L
120
4.635
−17.97
30.869
1
25.53
L


ATOM
910
CA
PRO
L
120
4.473
−17.362
28.532
1
25.8
L


ATOM
911
CB
PRO
L
120
5.414
−16.428
29.287
1
25.5
L


ATOM
912
CG
PRO
L
120
4.899
−16.495
30.691
1
27.71
L


ATOM
913
C
PRO
L
120
5.189
−18.077
27.384
1
25.45
L


ATOM
914
O
PRO
L
120
5.877
−19.085
27.586
1
26.64
L


ATOM
915
N
SER
L
121
5.024
−17.566
26.177
1
24.25
L


ATOM
916
CA
SER
L
121
5.686
−18.17
25.038
1
23.53
L


ATOM
917
CB
SER
L
121
5.119
−17.606
23.735
1
21.46
L


ATOM
918
OG
SER
L
121
5.214
−16.19
23.711
1
24.99
L


ATOM
919
C
SER
L
121
7.185
−17.886
25.098
1
23.69
L


ATOM
920
O
SER
L
121
7.625
−16.888
25.665
1
21.3
L


ATOM
921
N
SER
L
122
7.964
−18.789
24.524
1
26
L


ATOM
922
CA
SER
L
122
9.399
−18.622
24.456
1
25.49
L


ATOM
923
CB
SER
L
122
9.995
−19.783
23.644
1
27.93
L


ATOM
924
OG
SER
L
122
11.416
−19.771
23.635
1
34.44
L


ATOM
925
C
SER
L
122
9.593
−17.281
23.727
1
24.57
L


ATOM
926
O
SER
L
122
10.448
−16.472
24.091
1
23.46
L


ATOM
927
N
GLU
L
123
8.758
−17.052
22.715
1
24.73
L


ATOM
928
CA
GLU
L
123
8.8
−15.839
21.903
1
26.2
L


ATOM
929
CB
GLU
L
123
7.687
−15.849
20.849
1
26.46
L


ATOM
930
CG
GLU
L
123
7.91
−16.791
19.692
1
26.81
L


ATOM
931
CD
GLU
L
123
7.329
−18.158
19.941
1
29.3
L


ATOM
932
OE1
GLU
L
123
7.65
−18.768
20.985
1
32.39
L


ATOM
933
OE2
GLU
L
123
6.549
−18.627
19.088
1
31.33
L


ATOM
934
C
GLU
L
123
8.69
−14.538
22.683
1
27.76
L


ATOM
935
O
GLU
L
123
9.424
−13.579
22.414
1
30
L


ATOM
936
N
GLN
L
124
7.765
−14.495
23.637
1
27.59
L


ATOM
937
CA
GLN
L
124
7.551
−13.287
24.426
1
26.21
L


ATOM
938
CB
GLN
L
124
6.161
−13.325
25.061
1
25.73
L


ATOM
939
CG
GLN
L
124
5.878
−12.157
25.978
1
25.41
L


ATOM
940
CD
GLN
L
124
4.564
−12.31
26.7
1
28.18
L


ATOM
941
OE1
GLN
L
124
4.194
−13.414
27.104
1
30.55
L


ATOM
942
NE2
GLN
L
124
3.853
−11.202
26.883
1
29.49
L


ATOM
943
C
GLN
L
124
8.612
−13.057
25.498
1
24.05
L


ATOM
944
O
GLN
L
124
9.081
−11.937
25.68
1
20.76
L


ATOM
945
N
LEU
L
125
8.975
−14.117
26.21
1
25.83
L


ATOM
946
CA
LEU
L
125
9.985
−14.039
27.259
1
27.51
L


ATOM
947
CB
LEU
L
125
10.244
−15.429
27.829
1
26.43
L


ATOM
948
CG
LEU
L
125
9.11
−15.942
28.714
1
27.18
L


ATOM
949
CD1
LEU
L
125
9.232
−17.439
28.926
1
24.94
L


ATOM
950
CD2
LEU
L
125
9.144
−15.179
30.037
1
28.78
L


ATOM
951
C
LEU
L
125
11.265
−13.471
26.684
1
29.33
L


ATOM
952
O
LEU
L
125
11.957
−12.69
27.32
1
27.27
L


ATOM
953
N
THR
L
126
11.565
−13.883
25.462
1
34.71
L


ATOM
954
CA
THR
L
126
12.744
−13.422
24.747
1
38.58
L


ATOM
955
CB
THR
L
126
12.898
−14.217
23.418
1
38.73
L


ATOM
956
OG1
THR
L
126
13.508
−15.485
23.694
1
41.44
L


ATOM
957
CG2
THR
L
126
13.747
−13.456
22.415
1
42.63
L


ATOM
958
C
THR
L
126
12.649
−11.918
24.454
1
37.67
L


ATOM
959
O
THR
L
126
13.661
−11.244
24.277
1
38.38
L


ATOM
960
N
SER
L
127
11.424
−11.404
24.424
1
38.31
L


ATOM
961
CA
SER
L
127
11.174
−9.999
24.126
1
38.16
L


ATOM
962
CB
SER
L
127
9.895
−9.873
23.287
1
39.09
L


ATOM
963
OG
SER
L
127
9.902
−10.772
22.184
1
37.74
L


ATOM
964
C
SER
L
127
11.053
−9.124
25.373
1
38.2
L


ATOM
965
O
SER
L
127
10.884
−7.912
25.267
1
39.47
L


ATOM
966
N
GLY
L
128
11.131
−9.736
26.55
1
38.8
L


ATOM
967
CA
GLY
L
128
11.028
−8.974
27.784
1
36.79
L


ATOM
968
C
GLY
L
128
9.684
−9.089
28.483
1
36.17
L


ATOM
969
O
GLY
L
128
9.528
−8.655
29.628
1
37.33
L


ATOM
970
N
GLY
L
129
8.704
−9.676
27.807
1
33.66
L


ATOM
971
CA
GLY
L
129
7.396
−9.811
28.415
1
32.96
L


ATOM
972
C
GLY
L
129
7.086
−11.202
28.933
1
33.4
L


ATOM
973
O
GLY
L
129
7.796
−12.165
28.639
1
35.68
L


ATOM
974
N
ALA
L
130
6.021
−11.305
29.72
1
32.57
L


ATOM
975
CA
ALA
L
130
5.59
−12.585
30.271
1
29.83
L


ATOM
976
CB
ALA
L
130
6.379
−12.917
31.528
1
25.9
L


ATOM
977
C
ALA
L
130
4.095
−12.524
30.579
1
30.21
L


ATOM
978
O
ALA
L
130
3.649
−11.822
31.491
1
29.97
L


ATOM
979
N
SER
L
131
3.319
−13.253
29.794
1
28.89
L


ATOM
980
CA
SER
L
131
1.885
−13.283
29.991
1
28.16
L


ATOM
981
CB
SER
L
131
1.16
−12.762
28.748
1
27.57
L


ATOM
982
OG
SER
L
131
1.43
−11.384
28.549
1
28.65
L


ATOM
983
C
SER
L
131
1.462
−14.7
30.259
1
26.48
L


ATOM
984
O
SER
L
131
1.884
−15.622
29.571
1
29.19
L


ATOM
985
N
VAL
L
132
0.651
−14.886
31.284
1
25.94
L


ATOM
986
CA
VAL
L
132
0.159
−16.217
31.579
1
26.65
L


ATOM
987
CB
VAL
L
132
0.316
−16.551
33.066
1
26.59
L


ATOM
988
CG1
VAL
L
132
−0.139
−17.966
33.322
1
28.59
L


ATOM
989
CG2
VAL
L
132
1.771
−16.413
33.46
1
28.88
L


ATOM
990
C
VAL
L
132
−1.312
−16.234
31.159
1
25.12
L


ATOM
991
O
VAL
L
132
−2.047
−15.268
31.38
1
22.4
L


ATOM
992
N
VAL
L
133
−1.737
−17.324
30.534
1
23.9
L


ATOM
993
CA
VAL
L
133
−3.106
−17.4
30.058
1
23.63
L


ATOM
994
CB
VAL
L
133
−3.149
−17.498
28.531
1
18.35
L


ATOM
995
CG1
VAL
L
133
−4.592
−17.507
28.056
1
16.02
L


ATOM
996
CG2
VAL
L
133
−2.379
−16.339
27.922
1
16.76
L


ATOM
997
C
VAL
L
133
−3.952
−18.528
30.609
1
26.12
L


ATOM
998
O
VAL
L
133
−3.487
−19.65
30.816
1
27.97
L


ATOM
999
N
CYS
L
134
−5.223
−18.216
30.823
1
27.09
L


ATOM
1000
CA
CYS
L
134
−6.155
−19.197
31.326
1
24.04
L


ATOM
1001
C
CYS
L
134
−7.442
−19.179
30.513
1
20.92
L


ATOM
1002
O
CYS
L
134
−8.063
−18.133
30.339
1
17.8
L


ATOM
1003
CB
CYS
L
134
−6.463
−18.91
32.786
1
27.07
L


ATOM
1004
SG
CYS
L
134
−7.308
−20.273
33.647
1
32.1
L


ATOM
1005
N
PHE
L
135
−7.809
−20.334
29.977
1
18.32
L


ATOM
1006
CA
PHE
L
135
−9.05
−20.447
29.238
1
22.12
L


ATOM
1007
CB
PHE
L
135
−8.859
−21.182
27.905
1
23.68
L


ATOM
1008
CG
PHE
L
135
−8.09
−20.412
26.875
1
24.88
L


ATOM
1009
CD1
PHE
L
135
−8.396
−19.085
26.598
1
24.99
L


ATOM
1010
CD2
PHE
L
135
−7.055
−21.023
26.168
1
25.75
L


ATOM
1011
CE1
PHE
L
135
−7.675
−18.376
25.632
1
26.43
L


ATOM
1012
CE2
PHE
L
135
−6.328
−20.326
25.201
1
23.36
L


ATOM
1013
CZ
PHE
L
135
−6.636
−19.003
24.932
1
24.32
L


ATOM
1014
C
PHE
L
135
−10.025
−21.259
30.093
1
24.84
L


ATOM
1015
O
PHE
L
135
−9.694
−22.349
30.569
1
23.35
L


ATOM
1016
N
LEU
L
136
−11.211
−20.707
30.319
1
25.26
L


ATOM
1017
CA
LEU
L
136
−12.259
−21.407
31.056
1
27.46
L


ATOM
1018
CB
LEU
L
136
−12.831
−20.515
32.156
1
28.39
L


ATOM
1019
CG
LEU
L
136
−11.758
−19.815
32.994
1
27.24
L


ATOM
1020
CD1
LEU
L
136
−12.352
−19.383
34.319
1
29.28
L


ATOM
1021
CD2
LEU
L
136
−10.598
−20.749
33.234
1
30.57
L


ATOM
1022
C
LEU
L
136
−13.255
−21.606
29.926
1
28.23
L


ATOM
1023
O
LEU
L
136
−13.909
−20.657
29.487
1
28.24
L


ATOM
1024
N
ASN
L
137
−13.356
−22.836
29.436
1
28.97
L


ATOM
1025
CA
ASN
L
137
−14.207
−23.09
28.285
1
29.74
L


ATOM
1026
CB
ASN
L
137
−13.336
−23.608
27.155
1
28.72
L


ATOM
1027
CG
ASN
L
137
−12.318
−22.595
26.72
1
28.86
L


ATOM
1028
OD1
ASN
L
137
−12.602
−21.716
25.909
1
26.84
L


ATOM
1029
ND2
ASN
L
137
−11.125
−22.69
27.281
1
31.61
L


ATOM
1030
C
ASN
L
137
−15.425
−23.972
28.406
1
30.68
L


ATOM
1031
O
ASN
L
137
−15.478
−24.895
29.219
1
34.35
L


ATOM
1032
N
ASN
L
138
−16.396
−23.672
27.551
1
27.83
L


ATOM
1033
CA
ASN
L
138
−17.651
−24.399
27.48
1
27.16
L


ATOM
1034
CB
ASN
L
138
−17.44
−25.751
26.788
1
26.26
L


ATOM
1035
CG
ASN
L
138
−16.718
−25.615
25.455
1
25.93
L


ATOM
1036
OD1
ASN
L
138
−15.494
−25.5
25.41
1
26.96
L


ATOM
1037
ND2
ASN
L
138
−17.476
−25.609
24.365
1
24.51
L


ATOM
1038
C
ASN
L
138
−18.297
−24.602
28.839
1
26.32
L


ATOM
1039
O
ASN
L
138
−18.376
−25.717
29.348
1
26.78
L


ATOM
1040
N
PHE
L
139
−18.759
−23.509
29.43
1
25.31
L


ATOM
1041
CA
PHE
L
139
−19.416
−23.585
30.722
1
24.07
L


ATOM
1042
CB
PHE
L
139
−18.509
−23.036
31.818
1
19.54
L


ATOM
1043
CG
PHE
L
139
−18.23
−21.566
31.7
1
20.07
L


ATOM
1044
CD1
PHE
L
139
−19.147
−20.627
32.162
1
18.64
L


ATOM
1045
CD2
PHE
L
139
−17.039
−21.118
31.137
1
20.02
L


ATOM
1046
CE1
PHE
L
139
−18.879
−19.267
32.071
1
17.09
L


ATOM
1047
CE2
PHE
L
139
−16.764
−19.763
31.042
1
19.25
L


ATOM
1048
CZ
PHE
L
139
−17.687
−18.834
31.512
1
19.5
L


ATOM
1049
C
PHE
L
139
−20.705
−22.787
30.676
1
23.85
L


ATOM
1050
O
PHE
L
139
−20.896
−21.94
29.804
1
21.84
L


ATOM
1051
N
TYR
L
140
−21.592
−23.076
31.615
1
23.69
L


ATOM
1052
CA
TYR
L
140
−22.854
−22.37
31.699
1
25.1
L


ATOM
1053
CB
TYR
L
140
−23.862
−22.977
30.724
1
23.87
L


ATOM
1054
CG
TYR
L
140
−25.156
−22.205
30.639
1
26.42
L


ATOM
1055
CD1
TYR
L
140
−26.198
−22.452
31.534
1
25.3
L


ATOM
1056
CE1
TYR
L
140
−27.381
−21.728
31.477
1
24.72
L


ATOM
1057
CD2
TYR
L
140
−25.334
−21.206
29.68
1
26.48
L


ATOM
1058
CE2
TYR
L
140
−26.516
−20.474
29.615
1
25.77
L


ATOM
1059
CZ
TYR
L
140
−27.535
−20.743
30.517
1
27.81
L


ATOM
1060
OH
TYR
L
140
−28.715
−20.036
30.457
1
33.77
L


ATOM
1061
C
TYR
L
140
−23.343
−22.482
33.137
1
26.07
L


ATOM
1062
O
TYR
L
140
−23.196
−23.538
33.77
1
25.34
L


ATOM
1063
N
PRO
L
141
−23.918
−21.393
33.681
1
25.3
L


ATOM
1064
CD
PRO
L
141
−24.375
−21.396
35.085
1
25.57
L


ATOM
1065
CA
PRO
L
141
−24.154
−20.076
33.07
1
25.39
L


ATOM
1066
CB
PRO
L
141
−25.164
−19.447
34.018
1
24.54
L


ATOM
1067
CG
PRO
L
141
−24.663
−19.922
35.344
1
24.38
L


ATOM
1068
C
PRO
L
141
−22.908
−19.199
32.897
1
26.25
L


ATOM
1069
O
PRO
L
141
−21.828
−19.53
33.387
1
26.01
L


ATOM
1070
N
LYS
L
142
−23.087
−18.072
32.206
1
27.27
L


ATOM
1071
CA
LYS
L
142
−22.017
−17.117
31.914
1
27.69
L


ATOM
1072
CB
LYS
L
142
−22.593
−15.898
31.191
1
30.96
L


ATOM
1073
CG
LYS
L
142
−21.564
−14.897
30.677
1
33.13
L


ATOM
1074
CD
LYS
L
142
−22.264
−13.639
30.169
1
38.48
L


ATOM
1075
CE
LYS
L
142
−21.328
−12.707
29.408
1
41.44
L


ATOM
1076
NZ
LYS
L
142
−21.046
−13.194
28.018
1
45.59
L


ATOM
1077
C
LYS
L
142
−21.293
−16.647
33.161
1
28.47
L


ATOM
1078
O
LYS
L
142
−20.081
−16.423
33.15
1
31.54
L


ATOM
1079
N
ASP
L
143
−22.038
−16.485
34.24
1
28.45
L


ATOM
1080
CA
ASP
L
143
−21.446
−16.029
35.482
1
29.7
L


ATOM
1081
CB
ASP
L
143
−22.511
−15.959
36.569
1
33.15
L


ATOM
1082
CG
ASP
L
143
−21.982
−15.368
37.852
1
40.2
L


ATOM
1083
OD1
ASP
L
143
−21.041
−15.944
38.444
1
46.49
L


ATOM
1084
OD2
ASP
L
143
−22.505
−14.319
38.271
1
46.81
L


ATOM
1085
C
ASP
L
143
−20.304
−16.935
35.936
1
28.58
L


ATOM
1086
O
ASP
L
143
−20.479
−18.139
36.116
1
28.93
L


ATOM
1087
N
ILE
L
144
−19.129
−16.349
36.12
1
27.19
L


ATOM
1088
CA
ILE
L
144
−17.973
−17.107
36.571
1
25.59
L


ATOM
1089
CB
ILE
L
144
−17.359
−17.946
35.426
1
24.34
L


ATOM
1090
CG2
ILE
L
144
−16.577
−17.053
34.467
1
20.3
L


ATOM
1091
CG1
ILE
L
144
−16.447
−19.02
36.016
1
22.26
L


ATOM
1092
CD1
ILE
L
144
−15.87
−19.955
34.987
1
20.56
L


ATOM
1093
C
ILE
L
144
−16.944
−16.125
37.098
1
26.4
L


ATOM
1094
O
ILE
L
144
−16.891
−14.976
36.668
1
25.51
L


ATOM
1095
N
ASN
L
145
−16.13
−16.571
38.037
1
26.05
L


ATOM
1096
CA
ASN
L
145
−15.144
−15.69
38.612
1
30
L


ATOM
1097
CB
ASN
L
145
−15.607
−15.247
39.996
1
34.32
L


ATOM
1098
CG
ASN
L
145
−14.724
−14.174
40.582
1
43.24
L


ATOM
1099
OD1
ASN
L
145
−13.553
−14.421
40.894
1
48.53
L


ATOM
1100
ND2
ASN
L
145
−15.271
−12.965
40.728
1
41.97
L


ATOM
1101
C
ASN
L
145
−13.82
−16.411
38.693
1
30.84
L


ATOM
1102
O
ASN
L
145
−13.729
−17.499
39.249
1
34.14
L


ATOM
1103
N
VAL
L
146
−12.785
−15.807
38.133
1
30.13
L


ATOM
1104
CA
VAL
L
146
−11.481
−16.443
38.15
1
30.11
L


ATOM
1105
CB
VAL
L
146
−10.989
−16.699
36.716
1
29.97
L


ATOM
1106
CG1
VAL
L
146
−10.938
−15.4
35.956
1
29.52
L


ATOM
1107
CG2
VAL
L
146
−9.623
−17.351
36.743
1
27.67
L


ATOM
1108
C
VAL
L
146
−10.429
−15.624
38.882
1
29.94
L


ATOM
1109
O
VAL
L
146
−10.411
−14.398
38.793
1
29.16
L


ATOM
1110
N
LYS
L
147
−9.562
−16.302
39.624
1
28.51
L


ATOM
1111
CA
LYS
L
147
−8.498
−15.604
40.318
1
29.5
L


ATOM
1112
CB
LYS
L
147
−8.765
−15.514
41.824
1
31.78
L


ATOM
1113
CG
LYS
L
147
−8.653
−16.816
42.597
1
37.8
L


ATOM
1114
CD
LYS
L
147
−8.961
−16.584
44.078
1
42.9
L


ATOM
1115
CE
LYS
L
147
−10.364
−15.973
44.267
1
45.51
L


ATOM
1116
NZ
LYS
L
147
−10.704
−15.665
45.69
1
44.32
L


ATOM
1117
C
LYS
L
147
−7.191
−16.323
40.056
1
29.15
L


ATOM
1118
O
LYS
L
147
−7.163
−17.54
39.845
1
29.01
L


ATOM
1119
N
TRP
L
148
−6.111
−15.555
40.034
1
27.93
L


ATOM
1120
CA
TRP
L
148
−4.791
−16.11
39.798
1
25.36
L


ATOM
1121
CB
TRP
L
148
−3.979
−15.218
38.855
1
23.96
L


ATOM
1122
CG
TRP
L
148
−4.46
−15.205
37.44
1
25.11
L


ATOM
1123
CD2
TRP
L
148
−3.937
−15.979
36.357
1
24.69
L


ATOM
1124
CE2
TRP
L
148
−4.7
−15.665
35.209
1
25.65
L


ATOM
1125
CE3
TRP
L
148
−2.898
−16.908
36.244
1
20.89
L


ATOM
1126
CD1
TRP
L
148
−5.489
−14.472
36.924
1
25.85
L


ATOM
1127
NE1
TRP
L
148
−5.64
−14.743
35.583
1
27.1
L


ATOM
1128
CZ2
TRP
L
148
−4.455
−16.249
33.961
1
27.55
L


ATOM
1129
CZ3
TRP
L
148
−2.654
−17.487
35.008
1
24.29
L


ATOM
1130
CH2
TRP
L
148
−3.43
−17.156
33.879
1
23.39
L


ATOM
1131
C
TRP
L
148
−4.062
−16.198
41.119
1
25.07
L


ATOM
1132
O
TRP
L
148
−4.217
−15.337
41.982
1
27.06
L


ATOM
1133
N
LYS
L
149
−3.28
−17.248
41.289
1
21.62
L


ATOM
1134
CA
LYS
L
149
−2.512
−17.386
42.5
1
23.23
L


ATOM
1135
CB
LYS
L
149
−3.093
−18.473
43.398
1
25.26
L


ATOM
1136
CG
LYS
L
149
−4.325
−18.05
44.176
1
29.81
L


ATOM
1137
CD
LYS
L
149
−4.696
−19.134
45.173
1
33.68
L


ATOM
1138
CE
LYS
L
149
−5.822
−18.71
46.084
1
32.5
L


ATOM
1139
NZ
LYS
L
149
−6.016
−19.734
47.149
1
37.48
L


ATOM
1140
C
LYS
L
149
−1.102
−17.74
42.09
1
22.55
L


ATOM
1141
O
LYS
L
149
−0.89
−18.682
41.331
1
24.56
L


ATOM
1142
N
ILE
L
150
−0.141
−16.96
42.564
1
21.76
L


ATOM
1143
CA
ILE
L
150
1.257
−17.214
42.256
1
21.17
L


ATOM
1144
CB
ILE
L
150
1.943
−15.949
41.716
1
19.92
L


ATOM
1145
CG2
ILE
L
150
3.425
−16.201
41.534
1
23.07
L


ATOM
1146
CG1
ILE
L
150
1.287
−15.548
40.388
1
17.1
L


ATOM
1147
CD1
ILE
L
150
1.818
−14.283
39.765
1
15.75
L


ATOM
1148
C
ILE
L
150
1.89
−17.659
43.555
1
22.72
L


ATOM
1149
O
ILE
L
150
1.857
−16.94
44.55
1
22.25
L


ATOM
1150
N
ASP
L
151
2.449
−18.863
43.547
1
26.06
L


ATOM
1151
CA
ASP
L
151
3.046
−19.426
44.745
1
27.39
L


ATOM
1152
CB
ASP
L
151
4.395
−18.778
45.042
1
28.35
L


ATOM
1153
CG
ASP
L
151
5.475
−19.246
44.09
1
30.41
L


ATOM
1154
OD1
ASP
L
151
5.454
−20.438
43.718
1
31.21
L


ATOM
1155
OD2
ASP
L
151
6.349
−18.438
43.717
1
34.1
L


ATOM
1156
C
ASP
L
151
2.096
−19.236
45.907
1
29.88
L


ATOM
1157
O
ASP
L
151
2.453
−18.65
46.928
1
31.68
L


ATOM
1158
N
GLY
L
152
0.867
−19.714
45.713
1
34
L


ATOM
1159
CA
GLY
L
152
−0.163
−19.644
46.737
1
37.59
L


ATOM
1160
C
GLY
L
152
−0.932
−18.353
46.977
1
39.86
L


ATOM
1161
O
GLY
L
152
−2.115
−18.402
47.347
1
41.89
L


ATOM
1162
N
SER
L
153
−0.284
−17.206
46.779
1
38.11
L


ATOM
1163
CA
SER
L
153
−0.93
−15.92
47.019
1
37.57
L


ATOM
1164
CB
SER
L
153
0.083
−14.952
47.64
1
41.4
L


ATOM
1165
OG
SER
L
153
1.233
−14.8
46.821
1
47.97
L


ATOM
1166
C
SER
L
153
−1.602
−15.283
45.795
1
36.54
L


ATOM
1167
O
SER
L
153
−1.01
−15.166
44.719
1
33.76
L


ATOM
1168
N
GLU
L
154
−2.847
−14.857
45.986
1
36.02
L


ATOM
1169
CA
GLU
L
154
−3.638
−14.249
44.922
1
36.77
L


ATOM
1170
CB
GLU
L
154
−5.065
−13.974
45.405
1
37.67
L


ATOM
1171
CG
GLU
L
154
−5.894
−13.178
44.399
1
43.6
L


ATOM
1172
CD
GLU
L
154
−7.238
−12.738
44.944
1
47.14
L


ATOM
1173
OE1
GLU
L
154
−7.272
−12.138
46.04
1
52.56
L


ATOM
1174
OE2
GLU
L
154
−8.262
−12.979
44.273
1
50.9
L


ATOM
1175
C
GLU
L
154
−3.072
−12.96
44.353
1
34.65
L


ATOM
1176
O
GLU
L
154
−2.561
−12.121
45.084
1
36.38
L


ATOM
1177
N
ARG
L
155
−3.189
−12.811
43.038
1
33.44
L


ATOM
1178
CA
ARG
L
155
−2.733
−11.618
42.343
1
32.29
L


ATOM
1179
CB
ARG
L
155
−1.44
−11.88
41.583
1
29.56
L


ATOM
1180
CG
ARG
L
155
−1.085
−10.751
40.643
1
27.75
L


ATOM
1181
CD
ARG
L
155
0.387
−10.718
40.331
1
28.16
L


ATOM
1182
NE
ARG
L
155
0.7
−9.6
39.452
1
29.97
L


ATOM
1183
CZ
ARG
L
155
1.915
−9.085
39.307
1
32.61
L


ATOM
1184
NH1
ARG
L
155
2.942
−9.59
39.99
1
28.73
L


ATOM
1185
NH2
ARG
L
155
2.101
−8.06
38.483
1
32.32
L


ATOM
1186
C
ARG
L
155
−3.798
−11.163
41.361
1
33.97
L


ATOM
1187
O
ARG
L
155
−4.191
−11.914
40.464
1
34.48
L


ATOM
1188
N
GLN
L
156
−4.262
−9.93
41.527
1
35.27
L


ATOM
1189
CA
GLN
L
156
−5.289
−9.388
40.648
1
35.74
L


ATOM
1190
CB
GLN
L
156
−6.416
−8.752
41.469
1
34.74
L


ATOM
1191
CG
GLN
L
156
−6.968
−9.643
42.56
1
35.69
L


ATOM
1192
CD
GLN
L
156
−8.263
−9.112
43.152
1
39.08
L


ATOM
1193
OE1
GLN
L
156
−8.803
−9.684
44.105
1
38.61
L


ATOM
1194
NE2
GLN
L
156
−8.773
−8.018
42.586
1
37.2
L


ATOM
1195
C
GLN
L
156
−4.689
−8.343
39.722
1
36.18
L


ATOM
1196
O
GLN
L
156
−5.228
−8.067
38.65
1
37.15
L


ATOM
1197
N
ASN
L
157
−3.568
−7.767
40.14
1
35.69
L


ATOM
1198
CA
ASN
L
157
−2.904
−6.737
39.354
1
36.05
L


ATOM
1199
CB
ASN
L
157
−1.721
−6.159
40.138
1
35.1
L


ATOM
1200
CG
ASN
L
157
−1.028
−5.022
39.4
1
35.71
L


ATOM
1201
OD1
ASN
L
157
−1.67
−4.069
38.954
1
36.03
L


ATOM
1202
ND2
ASN
L
157
0.289
−5.114
39.279
1
35.37
L


ATOM
1203
C
ASN
L
157
−2.422
−7.265
38.016
1
35.99
L


ATOM
1204
O
ASN
L
157
−1.729
−8.277
37.956
1
39.57
L


ATOM
1205
N
GLY
L
158
−2.801
−6.579
36.942
1
36.3
L


ATOM
1206
CA
GLY
L
158
−2.381
−6.983
35.612
1
35.15
L


ATOM
1207
C
GLY
L
158
−3.187
−8.1
34.965
1
35.9
L


ATOM
1208
O
GLY
L
158
−2.68
−8.797
34.082
1
37.31
L


ATOM
1209
N
VAL
L
159
−4.44
−8.269
35.388
1
34.93
L


ATOM
1210
CA
VAL
L
159
−5.307
−9.302
34.834
1
31.19
L


ATOM
1211
CB
VAL
L
159
−6.04
−10.096
35.949
1
29.22
L


ATOM
1212
CG1
VAL
L
159
−6.974
−11.124
35.331
1
30.23
L


ATOM
1213
CG2
VAL
L
159
−5.04
−10.796
36.846
1
30.04
L


ATOM
1214
C
VAL
L
159
−6.358
−8.705
33.902
1
31.34
L


ATOM
1215
O
VAL
L
159
−6.862
−7.606
34.136
1
30.61
L


ATOM
1216
N
LEU
L
160
−6.67
−9.442
32.84
1
29.96
L


ATOM
1217
CA
LEU
L
160
−7.671
−9.036
31.863
1
31.26
L


ATOM
1218
CB
LEU
L
160
−7.01
−8.526
30.581
1
33.29
L


ATOM
1219
CG
LEU
L
160
−7.101
−7.018
30.349
1
36.88
L


ATOM
1220
CD1
LEU
L
160
−6.227
−6.301
31.361
1
37.95
L


ATOM
1221
CD2
LEU
L
160
−6.662
−6.678
28.933
1
41.51
L


ATOM
1222
C
LEU
L
160
−8.548
−10.237
31.543
1
29.31
L


ATOM
1223
O
LEU
L
160
−8.056
−11.312
31.202
1
31.42
L


ATOM
1224
N
ASN
L
161
−9.853
−10.049
31.645
1
27.25
L


ATOM
1225
CA
ASN
L
161
−10.783
−11.138
31.397
1
25.18
L


ATOM
1226
CB
ASN
L
161
−11.621
−11.375
32.648
1
23.62
L


ATOM
1227
CG
ASN
L
161
−10.765
−11.708
33.86
1
21.66
L


ATOM
1228
OD1
ASN
L
161
−11.118
−11.391
34.987
1
22.63
L


ATOM
1229
ND2
ASN
L
161
−9.642
−12.366
33.625
1
20.25
L


ATOM
1230
C
ASN
L
161
−11.672
−10.794
30.23
1
26.21
L


ATOM
1231
O
ASN
L
161
−12.17
−9.676
30.131
1
27.59
L


ATOM
1232
N
SER
L
162
−11.868
−11.761
29.344
1
25.37
L


ATOM
1233
CA
SER
L
162
−12.701
−11.555
28.177
1
25.42
L


ATOM
1234
CB
SER
L
162
−11.835
−11.356
26.932
1
23.65
L


ATOM
1235
OG
SER
L
162
−12.626
−10.895
25.852
1
25.84
L


ATOM
1236
C
SER
L
162
−13.628
−12.741
27.969
1
25.48
L


ATOM
1237
O
SER
L
162
−13.174
−13.884
27.879
1
25.86
L


ATOM
1238
N
TRP
L
163
−14.925
−12.458
27.9
1
25.51
L


ATOM
1239
CA
TRP
L
163
−15.942
−13.481
27.695
1
25.99
L


ATOM
1240
CB
TRP
L
163
−17.227
−13.107
28.43
1
27.7
L


ATOM
1241
CG
TRP
L
163
−17.326
−13.609
29.825
1
29.4
L


ATOM
1242
CD2
TRP
L
163
−16.764
−13.006
30.995
1
29.19
L


ATOM
1243
CE2
TRP
L
163
−17.132
−13.814
32.1
1
28.6
L


ATOM
1244
CE3
TRP
L
163
−15.985
−11.862
31.218
1
28.24
L


ATOM
1245
CD1
TRP
L
163
−17.991
−14.728
30.251
1
29.98
L


ATOM
1246
NE1
TRP
L
163
−17.88
−14.855
31.616
1
29.35
L


ATOM
1247
CZ2
TRP
L
163
−16.749
−13.515
33.407
1
26.04
L


ATOM
1248
CZ3
TRP
L
163
−15.602
−11.562
32.519
1
31.38
L


ATOM
1249
CH2
TRP
L
163
−15.987
−12.39
33.601
1
31.7
L


ATOM
1250
C
TRP
L
163
−16.272
−13.594
26.221
1
28.83
L


ATOM
1251
O
TRP
L
163
−16.162
−12.623
25.476
1
32.11
L


ATOM
1252
N
THR
L
164
−16.681
−14.782
25.806
1
27.08
L


ATOM
1253
CA
THR
L
164
−17.078
−15.006
24.434
1
27.43
L


ATOM
1254
CB
THR
L
164
−16.682
−16.405
23.983
1
28.7
L


ATOM
1255
OG1
THR
L
164
−17.458
−17.372
24.705
1
28.81
L


ATOM
1256
CG2
THR
L
164
−15.222
−16.654
24.269
1
31.49
L


ATOM
1257
C
THR
L
164
−18.602
−14.94
24.459
1
29.01
L


ATOM
1258
O
THR
L
164
−19.203
−14.85
25.526
1
28.21
L


ATOM
1259
N
ASP
L
165
−19.226
−14.965
23.289
1
31.53
L


ATOM
1260
CA
ASP
L
165
−20.681
−14.982
23.207
1
33.5
L


ATOM
1261
CB
ASP
L
165
−21.163
−14.457
21.85
1
41.28
L


ATOM
1262
CG
ASP
L
165
−21.484
−12.973
21.872
1
46.64
L


ATOM
1263
OD1
ASP
L
165
−22.521
−12.597
22.455
1
45.9
L


ATOM
1264
OD2
ASP
L
165
−20.698
−12.182
21.305
1
51.89
L


ATOM
1265
C
ASP
L
165
−21.006
−16.464
23.309
1
33.28
L


ATOM
1266
O
ASP
L
165
−20.099
−17.306
23.336
1
28.73
L


ATOM
1267
N
GLN
L
166
−22.285
−16.804
23.348
1
32.73
L


ATOM
1268
CA
GLN
L
166
−22.624
−18.211
23.435
1
35.31
L


ATOM
1269
CB
GLN
L
166
−24.115
−18.393
23.67
1
36.18
L


ATOM
1270
CG
GLN
L
166
−24.44
−18.734
25.094
1
37.34
L


ATOM
1271
CD
GLN
L
166
−25.92
−18.774
25.338
1
38.66
L


ATOM
1272
OE1
GLN
L
166
−26.635
−17.825
25.012
1
39.76
L


ATOM
1273
NE2
GLN
L
166
−26.397
−19.87
25.919
1
37.06
L


ATOM
1274
C
GLN
L
166
−22.194
−18.962
22.186
1
36.1
L


ATOM
1275
O
GLN
L
166
−22.494
−18.554
21.065
1
37.39
L


ATOM
1276
N
ASP
L
167
−21.486
−20.066
22.392
1
37.21
L


ATOM
1277
CA
ASP
L
167
−21.003
−20.877
21.29
1
37.98
L


ATOM
1278
CB
ASP
L
167
−19.97
−21.88
21.803
1
35.08
L


ATOM
1279
CG
ASP
L
167
−19.118
−22.442
20.692
1
32.96
L


ATOM
1280
OD1
ASP
L
167
−19.565
−23.384
20.013
1
32.35
L


ATOM
1281
OD2
ASP
L
167
−18.004
−21.921
20.487
1
33.42
L


ATOM
1282
C
ASP
L
167
−22.147
−21.6
20.572
1
39.75
L


ATOM
1283
O
ASP
L
167
−23.015
−22.209
21.198
1
37.53
L


ATOM
1284
N
SER
L
168
−22.128
−21.532
19.248
1
41
L


ATOM
1285
CA
SER
L
168
−23.165
−22.144
18.429
1
42.55
L


ATOM
1286
CB
SER
L
168
−23.013
−21.678
16.982
1
42.6
L


ATOM
1287
OG
SER
L
168
−23.892
−22.39
16.133
1
44.47
L


ATOM
1288
C
SER
L
168
−23.235
−23.669
18.445
1
43.61
L


ATOM
1289
O
SER
L
168
−24.193
−24.244
17.928
1
43.79
L


ATOM
1290
N
LYS
L
169
−22.238
−24.333
19.023
1
43.24
L


ATOM
1291
CA
LYS
L
169
−22.25
−25.792
19.04
1
42.34
L


ATOM
1292
CB
LYS
L
169
−20.822
−26.342
18.94
1
44.46
L


ATOM
1293
CG
LYS
L
169
−20.758
−27.822
18.57
1
47.38
L


ATOM
1294
CD
LYS
L
169
−19.368
−28.244
18.088
1
48.53
L


ATOM
1295
CE
LYS
L
169
−19.399
−29.643
17.459
1
50.07
L


ATOM
1296
NZ
LYS
L
169
−18.129
−30.007
16.751
1
47.84
L


ATOM
1297
C
LYS
L
169
−22.947
−26.372
20.265
1
41.98
L


ATOM
1298
O
LYS
L
169
−23.792
−27.256
20.132
1
45.03
L


ATOM
1299
N
ASP
L
170
−22.601
−25.879
21.451
1
38.81
L


ATOM
1300
CA
ASP
L
170
−23.208
−26.369
22.684
1
34.26
L


ATOM
1301
CB
ASP
L
170
−22.166
−27.092
23.532
1
34.62
L


ATOM
1302
CG
ASP
L
170
−20.945
−26.238
23.806
1
33.54
L


ATOM
1303
OD1
ASP
L
170
−21.023
−25.004
23.622
1
33.09
L


ATOM
1304
OD2
ASP
L
170
−19.911
−26.803
24.218
1
31.32
L


ATOM
1305
C
ASP
L
170
−23.829
−25.244
23.505
1
32.56
L


ATOM
1306
O
ASP
L
170
−24.205
−25.438
24.66
1
29.27
L


ATOM
1307
N
SER
L
171
−23.939
−24.072
22.895
1
30.66
L


ATOM
1308
CA
SER
L
171
−24.504
−22.898
23.551
1
32.6
L


ATOM
1309
CB
SER
L
171
−26.047
−22.996
23.596
1
34.23
L


ATOM
1310
OG
SER
L
171
−26.513
−24.277
23.983
1
38.27
L


ATOM
1311
C
SER
L
171
−23.929
−22.596
24.945
1
30.66
L


ATOM
1312
O
SER
L
171
−24.647
−22.188
25.861
1
28.18
L


ATOM
1313
N
THR
L
172
−22.619
−22.778
25.089
1
29.01
L


ATOM
1314
CA
THR
L
172
−21.947
−22.5
26.353
1
25.48
L


ATOM
1315
CB
THR
L
172
−20.963
−23.606
26.751
1
26.78
L


ATOM
1316
OG1
THR
L
172
−19.894
−23.643
25.799
1
27.48
L


ATOM
1317
CG2
THR
L
172
−21.653
−24.957
26.804
1
27.4
L


ATOM
1318
C
THR
L
172
−21.117
−21.248
26.183
1
23.04
L


ATOM
1319
O
THR
L
172
−20.908
−20.778
25.072
1
20.92
L


ATOM
1320
N
TYR
L
173
−20.636
−20.715
27.296
1
20.97
L


ATOM
1321
CA
TYR
L
173
−19.803
−19.541
27.25
1
22.45
L


ATOM
1322
CB
TYR
L
173
−20.231
−18.537
28.316
1
22.19
L


ATOM
1323
CG
TYR
L
173
−21.587
−17.901
28.079
1
26.18
L


ATOM
1324
CD1
TYR
L
173
−21.743
−16.838
27.186
1
21.86
L


ATOM
1325
CE1
TYR
L
173
−22.993
−16.256
26.971
1
22.12
L


ATOM
1326
CD2
TYR
L
173
−22.717
−18.368
28.752
1
24.39
L


ATOM
1327
CE2
TYR
L
173
−23.962
−17.799
28.546
1
24.98
L


ATOM
1328
CZ
TYR
L
173
−24.1
−16.744
27.659
1
25.53
L


ATOM
1329
OH
TYR
L
173
−25.351
−16.187
27.484
1
27.31
L


ATOM
1330
C
TYR
L
173
−18.385
−19.996
27.536
1
25.87
L


ATOM
1331
O
TYR
L
173
−18.164
−21.097
28.046
1
28.06
L


ATOM
1332
N
SER
L
174
−17.429
−19.145
27.191
1
24.94
L


ATOM
1333
CA
SER
L
174
−16.021
−19.402
27.441
1
23.7
L


ATOM
1334
CB
SER
L
174
−15.339
−19.973
26.194
1
24.79
L


ATOM
1335
OG
SER
L
174
−15.497
−21.378
26.126
1
22.74
L


ATOM
1336
C
SER
L
174
−15.401
−18.062
27.809
1
23.62
L


ATOM
1337
O
SER
L
174
−15.907
−17.005
27.429
1
21.15
L


ATOM
1338
N
MET
L
175
−14.324
−18.088
28.579
1
26.28
L


ATOM
1339
CA
MET
L
175
−13.676
−16.844
28.947
1
28.1
L


ATOM
1340
CB
MET
L
175
−14.152
−16.37
30.325
1
28.69
L


ATOM
1341
CG
MET
L
175
−13.07
−16.359
31.386
1
36.54
L


ATOM
1342
SD
MET
L
175
−13.289
−15.026
32.567
1
40.45
L


ATOM
1343
CE
MET
L
175
−11.621
−14.589
32.87
1
41.95
L


ATOM
1344
C
MET
L
175
−12.158
−16.974
28.915
1
27.05
L


ATOM
1345
O
MET
L
175
−11.599
−18.048
29.131
1
25.75
L


ATOM
1346
N
SER
L
176
−11.501
−15.862
28.63
1
27.02
L


ATOM
1347
CA
SER
L
176
−10.053
−15.826
28.556
1
27.71
L


ATOM
1348
CB
SER
L
176
−9.612
−15.335
27.175
1
25.91
L


ATOM
1349
OG
SER
L
176
−8.207
−15.348
27.048
1
28.02
L


ATOM
1350
C
SER
L
176
−9.542
−14.88
29.619
1
28.26
L


ATOM
1351
O
SER
L
176
−9.999
−13.746
29.72
1
28.42
L


ATOM
1352
N
SER
L
177
−8.594
−15.352
30.418
1
30.11
L


ATOM
1353
CA
SER
L
177
−8.012
−14.534
31.471
1
29.35
L


ATOM
1354
CB
SER
L
177
−8.288
−15.167
32.838
1
27.49
L


ATOM
1355
OG
SER
L
177
−7.717
−14.4
33.878
1
28.99
L


ATOM
1356
C
SER
L
177
−6.507
−14.432
31.231
1
31.17
L


ATOM
1357
O
SER
L
177
−5.808
−15.45
31.17
1
31.02
L


ATOM
1358
N
THR
L
178
−6.01
−13.208
31.079
1
31.47
L


ATOM
1359
CA
THR
L
178
−4.585
−12.998
30.845
1
31.49
L


ATOM
1360
CB
THR
L
178
−4.312
−12.274
29.518
1
29.61
L


ATOM
1361
OG1
THR
L
178
−4.819
−13.049
28.434
1
30.85
L


ATOM
1362
CG2
THR
L
178
−2.822
−12.1
29.316
1
30.54
L


ATOM
1363
C
THR
L
178
−3.949
−12.17
31.941
1
32
L


ATOM
1364
O
THR
L
178
−4.424
−11.086
32.263
1
32.95
L


ATOM
1365
N
LEU
L
179
−2.867
−12.7
32.503
1
33.31
L


ATOM
1366
CA
LEU
L
179
−2.111
−12.034
33.554
1
32.35
L


ATOM
1367
CB
LEU
L
179
−1.936
−12.973
34.739
1
30.55
L


ATOM
1368
CG
LEU
L
179
−0.966
−12.495
35.812
1
31.83
L


ATOM
1369
CD1
LEU
L
179
−1.346
−11.11
36.283
1
33.11
L


ATOM
1370
CD2
LEU
L
179
−0.987
−13.476
36.958
1
33.87
L


ATOM
1371
C
LEU
L
179
−0.744
−11.654
32.985
1
34.38
L


ATOM
1372
O
LEU
L
179
0.067
−12.523
32.649
1
33.53
L


ATOM
1373
N
THR
L
180
−0.49
−10.354
32.872
1
35.39
L


ATOM
1374
CA
THR
L
180
0.77
−9.881
32.319
1
36.62
L


ATOM
1375
CB
THR
L
180
0.533
−8.674
31.397
1
39.73
L


ATOM
1376
OG1
THR
L
180
−0.598
−8.937
30.555
1
43.88
L


ATOM
1377
CG2
THR
L
180
1.757
−8.423
30.519
1
38.15
L


ATOM
1378
C
THR
L
180
1.769
−9.495
33.403
1
36.52
L


ATOM
1379
O
THR
L
180
1.461
−8.72
34.306
1
35.29
L


ATOM
1380
N
LEU
L
181
2.972
−10.045
33.307
1
37.48
L


ATOM
1381
CA
LEU
L
181
4.013
−9.756
34.28
1
38.06
L


ATOM
1382
CB
LEU
L
181
4.33
−11.002
35.103
1
41.2
L


ATOM
1383
CG
LEU
L
181
3.182
−11.9
35.56
1
43.89
L


ATOM
1384
CD1
LEU
L
181
3.772
−13.128
36.213
1
45.68
L


ATOM
1385
CD2
LEU
L
181
2.265
−11.163
36.526
1
46.83
L


ATOM
1386
C
LEU
L
181
5.269
−9.343
33.536
1
38.17
L


ATOM
1387
O
LEU
L
181
5.323
−9.397
32.306
1
39.52
L


ATOM
1388
N
THR
L
182
6.279
−8.92
34.285
1
37.35
L


ATOM
1389
CA
THR
L
182
7.548
−8.552
33.681
1
35.94
L


ATOM
1390
CB
THR
L
182
8.337
−7.573
34.553
1
34.46
L


ATOM
1391
OG1
THR
L
182
8.89
−8.283
35.665
1
32.62
L


ATOM
1392
CG2
THR
L
182
7.43
−6.458
35.062
1
31.83
L


ATOM
1393
C
THR
L
182
8.314
−9.866
33.655
1
36.08
L


ATOM
1394
O
THR
L
182
8.057
−10.746
34.481
1
34.23
L


ATOM
1395
N
LYS
L
183
9.246
−10.004
32.716
1
35.5
L


ATOM
1396
CA
LYS
L
183
10.034
−11.225
32.623
1
34.49
L


ATOM
1397
CB
LYS
L
183
11.15
−11.055
31.595
1
33.98
L


ATOM
1398
CG
LYS
L
183
11.953
−12.317
31.341
1
31.95
L


ATOM
1399
CD
LYS
L
183
12.894
−12.128
30.166
1
32.27
L


ATOM
1400
CE
LYS
L
183
13.615
−13.415
29.817
1
33.24
L


ATOM
1401
NZ
LYS
L
183
14.518
−13.856
30.91
1
34.93
L


ATOM
1402
C
LYS
L
183
10.637
−11.575
33.982
1
35.2
L


ATOM
1403
O
LYS
L
183
10.611
−12.733
34.403
1
35.19
L


ATOM
1404
N
ASP
L
184
11.166
−10.568
34.669
1
33.82
L


ATOM
1405
CA
ASP
L
184
11.777
−10.778
35.972
1
34.87
L


ATOM
1406
CB
ASP
L
184
12.337
−9.467
36.512
1
39.39
L


ATOM
1407
CG
ASP
L
184
13.617
−9.066
35.828
1
44.5
L


ATOM
1408
OD1
ASP
L
184
14.481
−9.953
35.637
1
46.62
L


ATOM
1409
OD2
ASP
L
184
13.76
−7.871
35.492
1
47.09
L


ATOM
1410
C
ASP
L
184
10.842
−11.382
37.004
1
35.74
L


ATOM
1411
O
ASP
L
184
11.188
−12.381
37.644
1
35.61
L


ATOM
1412
N
GLU
L
185
9.672
−10.766
37.178
1
34.16
L


ATOM
1413
CA
GLU
L
185
8.682
−11.244
38.137
1
34.81
L


ATOM
1414
CB
GLU
L
185
7.383
−10.453
38.027
1
37.64
L


ATOM
1415
CG
GLU
L
185
7.409
−9.078
38.631
1
40.47
L


ATOM
1416
CD
GLU
L
185
6.124
−8.331
38.355
1
46.02
L


ATOM
1417
OE1
GLU
L
185
5.865
−8.011
37.168
1
46.14
L


ATOM
1418
OE2
GLU
L
185
5.367
−8.076
39.319
1
48.58
L


ATOM
1419
C
GLU
L
185
8.358
−12.698
37.881
1
34.43
L


ATOM
1420
O
GLU
L
185
8.452
−13.539
38.777
1
34.75
L


ATOM
1421
N
TYR
L
186
7.96
−12.981
36.647
1
32.69
L


ATOM
1422
CA
TYR
L
186
7.601
−14.332
36.254
1
30.01
L


ATOM
1423
CB
TYR
L
186
7.285
−14.375
34.757
1
28.03
L


ATOM
1424
CG
TYR
L
186
7.114
−15.773
34.206
1
23.69
L


ATOM
1425
CD1
TYR
L
186
6.049
−16.577
34.604
1
20.38
L


ATOM
1426
CE1
TYR
L
186
5.902
−17.866
34.115
1
21.84
L


ATOM
1427
CD2
TYR
L
186
8.031
−16.295
33.299
1
21.65
L


ATOM
1428
CE2
TYR
L
186
7.893
−17.577
32.801
1
23.26
L


ATOM
1429
CZ
TYR
L
186
6.826
−18.361
33.21
1
23.63
L


ATOM
1430
OH
TYR
L
186
6.685
−19.629
32.693
1
25.99
L


ATOM
1431
C
TYR
L
186
8.69
−15.347
36.578
1
29.28
L


ATOM
1432
O
TYR
L
186
8.389
−16.464
36.983
1
28.24
L


ATOM
1433
N
GLU
L
187
9.951
−14.962
36.421
1
29.32
L


ATOM
1434
CA
GLU
L
187
11.042
−15.889
36.687
1
31.9
L


ATOM
1435
CB
GLU
L
187
12.259
−15.514
35.843
1
35.89
L


ATOM
1436
CG
GLU
L
187
11.983
−15.468
34.352
1
41.42
L


ATOM
1437
CD
GLU
L
187
13.249
−15.31
33.531
1
46.71
L


ATOM
1438
OE1
GLU
L
187
14.167
−14.586
33.985
1
49.02
L


ATOM
1439
OE2
GLU
L
187
13.322
−15.896
32.426
1
47.81
L


ATOM
1440
C
GLU
L
187
11.466
−16.045
38.146
1
30.53
L


ATOM
1441
O
GLU
L
187
12.341
−16.844
38.448
1
30.22
L


ATOM
1442
N
ARG
L
188
10.858
−15.291
39.051
1
30.76
L


ATOM
1443
CA
ARG
L
188
11.216
−15.385
40.459
1
32.11
L


ATOM
1444
CB
ARG
L
188
11.141
−14.004
41.118
1
34.69
L


ATOM
1445
CG
ARG
L
188
12.121
−12.992
40.546
1
40.37
L


ATOM
1446
CD
ARG
L
188
13.537
−13.11
41.129
1
43.37
L


ATOM
1447
NE
ARG
L
188
14.184
−14.411
40.938
1
47.83
L


ATOM
1448
CZ
ARG
L
188
14.139
−15.416
41.815
1
47.96
L


ATOM
1449
NH1
ARG
L
188
13.473
−15.286
42.956
1
45.99
L


ATOM
1450
NH2
ARG
L
188
14.774
−16.551
41.557
1
47.26
L


ATOM
1451
C
ARG
L
188
10.289
−16.34
41.189
1
32.53
L


ATOM
1452
O
ARG
L
188
10.504
−16.664
42.356
1
33.78
L


ATOM
1453
N
HIS
L
189
9.266
−16.807
40.486
1
33.05
L


ATOM
1454
CA
HIS
L
189
8.282
−17.69
41.086
1
31.8
L


ATOM
1455
CB
HIS
L
189
6.939
−16.973
41.069
1
30.04
L


ATOM
1456
CG
HIS
L
189
6.96
−15.68
41.821
1
28.38
L


ATOM
1457
CD2
HIS
L
189
7.114
−14.403
41.395
1
27.92
L


ATOM
1458
ND1
HIS
L
189
6.897
−15.621
43.198
1
25.92
L


ATOM
1459
CE1
HIS
L
189
7.012
−14.363
43.586
1
29.08
L


ATOM
1460
NE2
HIS
L
189
7.146
−13.604
42.512
1
27.83
L


ATOM
1461
C
HIS
L
189
8.183
−19.083
40.467
1
33.49
L


ATOM
1462
O
HIS
L
189
8.781
−19.368
39.434
1
33.11
L


ATOM
1463
N
ASN
L
190
7.418
−19.951
41.113
1
33.72
L


ATOM
1464
CA
ASN
L
190
7.292
−21.312
40.647
1
34.37
L


ATOM
1465
CB
ASN
L
190
7.859
−22.254
41.707
1
39.62
L


ATOM
1466
CG
ASN
L
190
7.431
−23.686
41.488
1
45.15
L


ATOM
1467
OD1
ASN
L
190
7.872
−24.336
40.537
1
47.16
L


ATOM
1468
ND2
ASN
L
190
6.561
−24.188
42.364
1
44.99
L


ATOM
1469
C
ASN
L
190
5.886
−21.779
40.261
1
32.92
L


ATOM
1470
O
ASN
L
190
5.669
−22.211
39.126
1
32.41
L


ATOM
1471
N
SER
L
191
4.942
−21.726
41.2
1
29.03
L


ATOM
1472
CA
SER
L
191
3.583
−22.178
40.91
1
26.27
L


ATOM
1473
CB
SER
L
191
2.982
−22.937
42.091
1
27.46
L


ATOM
1474
OG
SER
L
191
1.622
−23.25
41.826
1
27.07
L


ATOM
1475
C
SER
L
191
2.601
−21.101
40.495
1
24.38
L


ATOM
1476
O
SER
L
191
2.327
−20.154
41.237
1
23.92
L


ATOM
1477
N
TYR
L
192
2.08
−21.268
39.287
1
20.25
L


ATOM
1478
CA
TYR
L
192
1.096
−20.367
38.734
1
20.14
L


ATOM
1479
CB
TYR
L
192
1.518
−19.919
37.342
1
19.68
L


ATOM
1480
CG
TYR
L
192
2.707
−19.005
37.396
1
19.45
L


ATOM
1481
CD1
TYR
L
192
3.999
−19.511
37.535
1
17.45
L


ATOM
1482
CE1
TYR
L
192
5.096
−18.654
37.658
1
21.02
L


ATOM
1483
CD2
TYR
L
192
2.536
−17.623
37.377
1
19.72
L


ATOM
1484
CE2
TYR
L
192
3.617
−16.761
37.497
1
21.17
L


ATOM
1485
CZ
TYR
L
192
4.892
−17.278
37.636
1
21.38
L


ATOM
1486
OH
TYR
L
192
5.951
−16.405
37.735
1
24.43
L


ATOM
1487
C
TYR
L
192
−0.23
−21.114
38.693
1
21.58
L


ATOM
1488
O
TYR
L
192
−0.367
−22.145
38.032
1
21.82
L


ATOM
1489
N
THR
L
193
−1.209
−20.588
39.409
1
20.4
L


ATOM
1490
CA
THR
L
193
−2.491
−21.243
39.485
1
24.31
L


ATOM
1491
CB
THR
L
193
−2.757
−21.705
40.925
1
26.79
L


ATOM
1492
OG1
THR
L
193
−1.554
−22.253
41.48
1
31.26
L


ATOM
1493
CG2
THR
L
193
−3.828
−22.76
40.954
1
25.88
L


ATOM
1494
C
THR
L
193
−3.628
−20.347
39.048
1
25.3
L


ATOM
1495
O
THR
L
193
−3.688
−19.173
39.397
1
26.69
L


ATOM
1496
N
CYS
L
194
−4.535
−20.923
38.279
1
25.93
L


ATOM
1497
CA
CYS
L
194
−5.703
−20.209
37.8
1
28.68
L


ATOM
1498
C
CYS
L
194
−6.9
−20.9
38.446
1
26.29
L


ATOM
1499
O
CYS
L
194
−7.132
−22.088
38.219
1
23.6
L


ATOM
1500
CB
CYS
L
194
−5.776
−20.302
36.275
1
30.58
L


ATOM
1501
SG
CYS
L
194
−7.316
−19.641
35.58
1
37.8
L


ATOM
1502
N
GLU
L
195
−7.645
−20.167
39.267
1
25.68
L


ATOM
1503
CA
GLU
L
195
−8.787
−20.757
39.951
1
24.58
L


ATOM
1504
CB
GLU
L
195
−8.678
−20.557
41.46
1
30.25
L


ATOM
1505
CG
GLU
L
195
−7.31
−20.86
42.031
1
33.55
L


ATOM
1506
CD
GLU
L
195
−7.349
−21.025
43.526
1
38.55
L


ATOM
1507
OE1
GLU
L
195
−8.034
−20.212
44.191
1
41.68
L


ATOM
1508
OE2
GLU
L
195
−6.695
−21.961
44.035
1
40.72
L


ATOM
1509
C
GLU
L
195
−10.101
−20.19
39.468
1
23.83
L


ATOM
1510
O
GLU
L
195
−10.289
−18.972
39.395
1
19.93
L


ATOM
1511
N
ALA
L
196
−11.012
−21.101
39.15
1
23.99
L


ATOM
1512
CA
ALA
L
196
−12.324
−20.738
38.654
1
23.37
L


ATOM
1513
CB
ALA
L
196
−12.597
−21.475
37.353
1
20.82
L


ATOM
1514
C
ALA
L
196
−13.408
−21.064
39.662
1
23.23
L


ATOM
1515
O
ALA
L
196
−13.475
−22.174
40.185
1
25.37
L


ATOM
1516
N
THR
L
197
−14.25
−20.084
39.946
1
24.13
L


ATOM
1517
CA
THR
L
197
−15.367
−20.288
40.851
1
22.79
L


ATOM
1518
CB
THR
L
197
−15.396
−19.231
41.964
1
23.35
L


ATOM
1519
OG1
THR
L
197
−14.312
−19.483
42.862
1
28.46
L


ATOM
1520
CG2
THR
L
197
−16.705
−19.293
42.746
1
24.14
L


ATOM
1521
C
THR
L
197
−16.599
−20.187
39.968
1
23.18
L


ATOM
1522
O
THR
L
197
−16.814
−19.182
39.288
1
20.44
L


ATOM
1523
N
HIS
L
198
−17.385
−21.256
39.962
1
24.07
L


ATOM
1524
CA
HIS
L
198
−18.579
−21.34
39.143
1
24.32
L


ATOM
1525
CB
HIS
L
198
−18.202
−21.968
37.803
1
25.38
L


ATOM
1526
CG
HIS
L
198
−19.321
−22.008
36.815
1
29.02
L


ATOM
1527
CD2
HIS
L
198
−19.631
−21.182
35.788
1
28.67
L


ATOM
1528
ND1
HIS
L
198
−20.294
−22.983
36.833
1
29.44
L


ATOM
1529
CE1
HIS
L
198
−21.157
−22.756
35.858
1
31.04
L


ATOM
1530
NE2
HIS
L
198
−20.778
−21.669
35.21
1
30.62
L


ATOM
1531
C
HIS
L
198
−19.603
−22.189
39.895
1
26.72
L


ATOM
1532
O
HIS
L
198
−19.236
−23.036
40.711
1
25.66
L


ATOM
1533
N
LYS
L
199
−20.884
−21.97
39.622
1
25.17
L


ATOM
1534
CA
LYS
L
199
−21.927
−22.696
40.332
1
25.25
L


ATOM
1535
CB
LYS
L
199
−23.293
−22.121
39.98
1
25.87
L


ATOM
1536
CG
LYS
L
199
−23.775
−22.463
38.59
1
25.86
L


ATOM
1537
CD
LYS
L
199
−25.151
−21.89
38.392
1
28.54
L


ATOM
1538
CE
LYS
L
199
−26.118
−22.43
39.436
1
30.74
L


ATOM
1539
NZ
LYS
L
199
−27.463
−21.792
39.348
1
33.3
L


ATOM
1540
C
LYS
L
199
−21.956
−24.204
40.127
1
26.62
L


ATOM
1541
O
LYS
L
199
−22.658
−24.907
40.85
1
28.56
L


ATOM
1542
N
THR
L
200
−21.205
−24.704
39.151
1
27.16
L


ATOM
1543
CA
THR
L
200
−21.176
−26.138
38.876
1
26.19
L


ATOM
1544
CB
THR
L
200
−20.77
−26.402
37.422
1
25.98
L


ATOM
1545
OG1
THR
L
200
−19.635
−25.595
37.094
1
26.58
L


ATOM
1546
CG2
THR
L
200
−21.919
−26.069
36.477
1
26.9
L


ATOM
1547
C
THR
L
200
−20.221
−26.893
39.8
1
27.16
L


ATOM
1548
O
THR
L
200
−20.068
−28.119
39.711
1
27.77
L


ATOM
1549
N
SER
L
201
−19.577
−26.153
40.691
1
25.31
L


ATOM
1550
CA
SER
L
201
−18.645
−26.747
41.63
1
24.1
L


ATOM
1551
CB
SER
L
201
−17.229
−26.682
41.078
1
20.07
L


ATOM
1552
OG
SER
L
201
−16.317
−27.119
42.061
1
18.73
L


ATOM
1553
C
SER
L
201
−18.689
−26.041
42.978
1
25.8
L


ATOM
1554
O
SER
L
201
−18.703
−24.813
43.053
1
23.37
L


ATOM
1555
N
THR
L
202
−18.717
−26.826
44.046
1
28.38
L


ATOM
1556
CA
THR
L
202
−18.74
−26.261
45.385
1
29.58
L


ATOM
1557
CB
THR
L
202
−19.066
−27.341
46.424
1
30.37
L


ATOM
1558
OG1
THR
L
202
−20.407
−27.803
46.217
1
33.81
L


ATOM
1559
CG2
THR
L
202
−18.931
−26.788
47.826
1
30.43
L


ATOM
1560
C
THR
L
202
−17.375
−25.647
45.685
1
28.79
L


ATOM
1561
O
THR
L
202
−17.274
−24.594
46.316
1
26.39
L


ATOM
1562
N
SER
L
203
−16.326
−26.303
45.209
1
28.25
L


ATOM
1563
CA
SER
L
203
−14.973
−25.823
45.426
1
31.48
L


ATOM
1564
CB
SER
L
203
−14.04
−26.985
45.75
1
30.8
L


ATOM
1565
OG
SER
L
203
−13.848
−27.801
44.603
1
35.58
L


ATOM
1566
C
SER
L
203
−14.467
−25.138
44.173
1
32.55
L


ATOM
1567
O
SER
L
203
−15.096
−25.197
43.121
1
34.09
L


ATOM
1568
N
PRO
L
204
−13.311
−24.476
44.271
1
32.91
L


ATOM
1569
CD
PRO
L
204
−12.652
−24.015
45.503
1
34.19
L


ATOM
1570
CA
PRO
L
204
−12.757
−23.796
43.103
1
29.91
L


ATOM
1571
CB
PRO
L
204
−11.681
−22.896
43.705
1
32.01
L


ATOM
1572
CG
PRO
L
204
−12.184
−22.64
45.097
1
33.94
L


ATOM
1573
C
PRO
L
204
−12.163
−24.812
42.144
1
29.65
L


ATOM
1574
O
PRO
L
204
−11.408
−25.7
42.558
1
28.69
L


ATOM
1575
N
ILE
L
205
−12.526
−24.688
40.872
1
27.52
L


ATOM
1576
CA
ILE
L
205
−12.014
−25.561
39.823
1
25.93
L


ATOM
1577
CB
ILE
L
205
−12.931
−25.553
38.608
1
24.84
L


ATOM
1578
CG2
ILE
L
205
−12.352
−26.442
37.528
1
22.4
L


ATOM
1579
CG1
ILE
L
205
−14.333
−25.996
39.027
1
22.64
L


ATOM
1580
CD1
ILE
L
205
−15.31
−26.084
37.893
1
27.42
L


ATOM
1581
C
ILE
L
205
−10.689
−24.931
39.442
1
27.85
L


ATOM
1582
O
ILE
L
205
−10.663
−23.817
38.908
1
27.35
L


ATOM
1583
N
VAL
L
206
−9.592
−25.641
39.693
1
29.18
L


ATOM
1584
CA
VAL
L
206
−8.279
−25.071
39.438
1
29.6
L


ATOM
1585
CB
VAL
L
206
−7.581
−24.788
40.783
1
31.17
L


ATOM
1586
CG1
VAL
L
206
−6.77
−26.007
41.228
1
28.47
L


ATOM
1587
CG2
VAL
L
206
−6.725
−23.558
40.663
1
32.18
L


ATOM
1588
C
VAL
L
206
−7.317
−25.858
38.556
1
29.44
L


ATOM
1589
O
VAL
L
206
−7.346
−27.087
38.523
1
33.05
L


ATOM
1590
N
LYS
L
207
−6.443
−25.135
37.862
1
26.67
L


ATOM
1591
CA
LYS
L
207
−5.449
−25.747
36.989
1
26.89
L


ATOM
1592
CB
LYS
L
207
−5.934
−25.682
35.546
1
30.37
L


ATOM
1593
CG
LYS
L
207
−5.217
−26.621
34.596
1
33.58
L


ATOM
1594
CD
LYS
L
207
−5.748
−28.03
34.722
1
35.21
L


ATOM
1595
CE
LYS
L
207
−5.121
−28.948
33.679
1
40.24
L


ATOM
1596
NZ
LYS
L
207
−3.641
−29.062
33.831
1
44.1
L


ATOM
1597
C
LYS
L
207
−4.124
−24.986
37.131
1
26.74
L


ATOM
1598
O
LYS
L
207
−4.121
−23.767
37.258
1
26.03
L


ATOM
1599
N
SER
L
208
−3
−25.694
37.096
1
28.03
L


ATOM
1600
CA
SER
L
208
−1.705
−25.034
37.239
1
30.63
L


ATOM
1601
CB
SER
L
208
−1.218
−25.113
38.701
1
32.54
L


ATOM
1602
OG
SER
L
208
−2.183
−24.625
39.623
1
35.32
L


ATOM
1603
C
SER
L
208
−0.611
−25.616
36.347
1
29.96
L


ATOM
1604
O
SER
L
208
−0.747
−26.708
35.793
1
29.65
L


ATOM
1605
N
PHE
L
209
0.461
−24.848
36.185
1
31.99
L


ATOM
1606
CA
PHE
L
209
1.641
−25.286
35.441
1
33.23
L


ATOM
1607
CB
PHE
L
209
1.782
−24.561
34.087
1
28.82
L


ATOM
1608
CG
PHE
L
209
2.181
−23.123
34.169
1
27.52
L


ATOM
1609
CD1
PHE
L
209
3.47
−22.758
34.552
1
26.04
L


ATOM
1610
CD2
PHE
L
209
1.297
−22.13
33.752
1
27.83
L


ATOM
1611
CE1
PHE
L
209
3.876
−21.42
34.509
1
25.58
L


ATOM
1612
CE2
PHE
L
209
1.692
−20.791
33.704
1
26.94
L


ATOM
1613
CZ
PHE
L
209
2.985
−20.436
34.081
1
25.94
L


ATOM
1614
C
PHE
L
209
2.699
−24.947
36.477
1
35.23
L


ATOM
1615
O
PHE
L
209
2.397
−24.216
37.418
1
33.97
L


ATOM
1616
N
ASN
L
210
3.928
−25.414
36.339
1
40.46
L


ATOM
1617
CA
ASN
L
210
4.809
−25.173
37.461
1
45.08
L


ATOM
1618
CB
ASN
L
210
4.696
−26.398
38.364
1
46.07
L


ATOM
1619
CG
ASN
L
210
4.953
−26.079
39.798
1
47.59
L


ATOM
1620
OD1
ASN
L
210
6.076
−26.213
40.281
1
52.21
L


ATOM
1621
ND2
ASN
L
210
3.912
−25.655
40.503
1
48.79
L


ATOM
1622
C
ASN
L
210
6.264
−24.76
37.397
1
48.6
L


ATOM
1623
O
ASN
L
210
7.071
−25.309
38.136
1
52.51
L


ATOM
1624
N
ARG
L
211
6.63
−23.81
36.553
1
50.38
L


ATOM
1625
CA
ARG
L
211
8.027
−23.381
36.531
1
55.12
L


ATOM
1626
CB
ARG
L
211
8.854
−24.314
35.647
1
56.21
L


ATOM
1627
CG
ARG
L
211
10.013
−25.016
36.385
1
54.65
L


ATOM
1628
CD
ARG
L
211
9.524
−26.033
37.398
1
56.33
L


ATOM
1629
NE
ARG
L
211
9.655
−27.429
36.963
1
58.85
L


ATOM
1630
CZ
ARG
L
211
9.003
−27.997
35.945
1
59.6
L


ATOM
1631
NH1
ARG
L
211
8.138
−27.309
35.206
1
60.35
L


ATOM
1632
NH2
ARG
L
211
9.228
−29.273
35.659
1
59.17
L


ATOM
1633
C
ARG
L
211
8.086
−21.948
36.016
1
59.17
L


ATOM
1634
O
ARG
L
211
7.1
−21.448
35.472
1
63.18
L


ATOM
1635
N
ASN
L
212
9.206
−21.263
36.227
1
62.81
L


ATOM
1636
CA
ASN
L
212
9.343
−19.884
35.751
1
64.64
L


ATOM
1637
CB
ASN
L
212
9.893
−18.979
36.856
1
64.51
L


ATOM
1638
CG
ASN
L
212
11.232
−19.443
37.385
1
65.7
L


ATOM
1639
OD1
ASN
L
212
11.303
−20.364
38.197
1
69.15
L


ATOM
1640
ND2
ASN
L
212
12.305
−18.809
36.927
1
66.81
L


ATOM
1641
C
ASN
L
212
10.256
−19.809
34.527
1
68.47
L


ATOM
1642
O
ASN
L
212
11.336
−19.187
34.621
1
70.6
L


ATOM
1643
OXT
ASN
L
212
9.884
−20.387
33.481
1
70.56
L


ATOM
1644
CB
GLN
H
1
−22.086
−14.508
−7.075
1
26.29
H


ATOM
1645
CG
GLN
H
1
−20.652
−14.421
−7.602
1
32.57
H


ATOM
1646
CD
GLN
H
1
−20.557
−14.579
−9.118
1
34.89
H


ATOM
1647
OE1
GLN
H
1
−21.038
−15.562
−9.679
1
37.29
H


ATOM
1648
NE2
GLN
H
1
−19.934
−13.606
−9.784
1
36.04
H


ATOM
1649
C
GLN
H
1
−22.835
−12.119
−7.279
1
21.84
H


ATOM
1650
O
GLN
H
1
−21.754
−11.559
−7.446
1
22.55
H


ATOM
1651
N
GLN
H
1
−22.88
−13.61
−9.234
1
25.61
H


ATOM
1652
CA
GLN
H
1
−23.062
−13.544
−7.756
1
24.49
H


ATOM
1653
N
VAL
H
2
−23.868
−11.52
−6.71
1
21.4
H


ATOM
1654
CA
VAL
H
2
−23.753
−10.155
−6.218
1
25.31
H


ATOM
1655
CB
VAL
H
2
−25.111
−9.613
−5.698
1
24.56
H


ATOM
1656
CG1
VAL
H
2
−24.905
−8.286
−4.997
1
22.03
H


ATOM
1657
CG2
VAL
H
2
−26.08
−9.436
−6.85
1
25.35
H


ATOM
1658
C
VAL
H
2
−22.765
−10.104
−5.065
1
27.12
H


ATOM
1659
O
VAL
H
2
−22.928
−10.809
−4.076
1
26.63
H


ATOM
1660
N
GLN
H
3
−21.735
−9.28
−5.198
1
31.38
H


ATOM
1661
CA
GLN
H
3
−20.761
−9.13
−4.13
1
35.8
H


ATOM
1662
CB
GLN
H
3
−19.685
−10.216
−4.223
1
37.26
H


ATOM
1663
CG
GLN
H
3
−19.196
−10.531
−5.609
1
42.81
H


ATOM
1664
CD
GLN
H
3
−18.323
−11.772
−5.617
1
46.01
H


ATOM
1665
OE1
GLN
H
3
−18.748
−12.846
−5.186
1
45.7
H


ATOM
1666
NE2
GLN
H
3
−17.096
−11.631
−6.101
1
47.01
H


ATOM
1667
C
GLN
H
3
−20.134
−7.736
−4.069
1
36.25
H


ATOM
1668
O
GLN
H
3
−19.987
−7.053
−5.086
1
34.29
H


ATOM
1669
N
LEU
H
4
−19.777
−7.328
−2.855
1
36.84
H


ATOM
1670
CA
LEU
H
4
−19.192
−6.016
−2.615
1
38.19
H


ATOM
1671
CB
LEU
H
4
−20.171
−5.176
−1.792
1
37.01
H


ATOM
1672
CG
LEU
H
4
−21.599
−5.129
−2.335
1
35.4
H


ATOM
1673
CD1
LEU
H
4
−22.525
−4.511
−1.302
1
32.9
H


ATOM
1674
CD2
LEU
H
4
−21.615
−4.341
−3.639
1
35.39
H


ATOM
1675
C
LEU
H
4
−17.846
−6.096
−1.887
1
38.77
H


ATOM
1676
O
LEU
H
4
−17.777
−6.489
−0.718
1
39.1
H


ATOM
1677
N
GLN
H
5
−16.78
−5.708
−2.579
1
37.65
H


ATOM
1678
CA
GLN
H
5
−15.457
−5.74
−1.986
1
38.61
H


ATOM
1679
CB
GLN
H
5
−14.459
−6.348
−2.975
1
41.32
H


ATOM
1680
CG
GLN
H
5
−13.209
−6.874
−2.299
1
51.07
H


ATOM
1681
CD
GLN
H
5
−13.536
−7.799
−1.128
1
56.81
H


ATOM
1682
OE1
GLN
H
5
−13.909
−8.962
−1.316
1
59.1
H


ATOM
1683
NE2
GLN
H
5
−13.413
−7.275
0.089
1
59.64
H


ATOM
1684
C
GLN
H
5
−15.035
−4.326
−1.574
1
36.03
H


ATOM
1685
O
GLN
H
5
−14.778
−3.474
−2.42
1
36.13
H


ATOM
1686
N
GLN
H
6
−14.974
−4.086
−0.267
1
33.5
H


ATOM
1687
CA
GLN
H
6
−14.607
−2.776
0.273
1
33.36
H


ATOM
1688
CB
GLN
H
6
−15.299
−2.551
1.619
1
30.65
H


ATOM
1689
CG
GLN
H
6
−16.799
−2.751
1.588
1
30.36
H


ATOM
1690
CD
GLN
H
6
−17.459
−2.383
2.905
1
30.69
H


ATOM
1691
OE1
GLN
H
6
−18.563
−2.844
3.211
1
29.76
H


ATOM
1692
NE2
GLN
H
6
−16.794
−1.539
3.685
1
26.25
H


ATOM
1693
C
GLN
H
6
−13.106
−2.609
0.471
1
33.62
H


ATOM
1694
O
GLN
H
6
−12.36
−3.589
0.46
1
35
H


ATOM
1695
N
SER
H
7
−12.663
−1.367
0.66
1
32.15
H


ATOM
1696
CA
SER
H
7
−11.247
−1.104
0.885
1
31.54
H


ATOM
1697
CB
SER
H
7
−10.939
0.39
0.754
1
33.62
H


ATOM
1698
OG
SER
H
7
−11.584
1.152
1.76
1
36.36
H


ATOM
1699
C
SER
H
7
−10.908
−1.593
2.288
1
31.21
H


ATOM
1700
O
SER
H
7
−11.802
−1.786
3.112
1
31.96
H


ATOM
1701
N
GLY
H
8
−9.625
−1.801
2.561
1
30.61
H


ATOM
1702
CA
GLY
H
8
−9.23
−2.289
3.872
1
29.48
H


ATOM
1703
C
GLY
H
8
−9.337
−1.294
5.017
1
30.07
H


ATOM
1704
O
GLY
H
8
−9.365
−0.08
4.799
1
26.82
H


ATOM
1705
N
SER
H
9
−9.398
−1.817
6.242
1
30.18
H


ATOM
1706
CA
SER
H
9
−9.484
−0.983
7.435
1
31.66
H


ATOM
1707
CB
SER
H
9
−9.394
−1.837
8.699
1
29.38
H


ATOM
1708
OG
SER
H
9
−10.459
−2.766
8.763
1
32.73
H


ATOM
1709
C
SER
H
9
−8.336
0.015
7.417
1
33.59
H


ATOM
1710
O
SER
H
9
−7.322
−0.209
6.753
1
32.55
H


ATOM
1711
N
GLU
H
10
−8.492
1.115
8.145
1
35.94
H


ATOM
1712
CA
GLU
H
10
−7.454
2.135
8.181
1
37.9
H


ATOM
1713
CB
GLU
H
10
−7.563
3.046
6.955
1
39.79
H


ATOM
1714
CG
GLU
H
10
−6.331
3.024
6.061
1
45.36
H


ATOM
1715
CD
GLU
H
10
−6.374
4.079
4.967
1
49.51
H


ATOM
1716
OE1
GLU
H
10
−7.365
4.12
4.197
1
51.22
H


ATOM
1717
OE2
GLU
H
10
−5.407
4.868
4.876
1
50.77
H


ATOM
1718
C
GLU
H
10
−7.483
2.995
9.433
1
37.37
H


ATOM
1719
O
GLU
H
10
−8.529
3.202
10.038
1
37.12
H


ATOM
1720
N
LEU
H
11
−6.31
3.488
9.812
1
38.36
H


ATOM
1721
CA
LEU
H
11
−6.162
4.358
10.969
1
38.49
H


ATOM
1722
CB
LEU
H
11
−4.981
3.899
11.828
1
39.9
H


ATOM
1723
CG
LEU
H
11
−4.625
4.687
13.093
1
42.26
H


ATOM
1724
CD1
LEU
H
11
−5.844
4.869
13.986
1
41.04
H


ATOM
1725
CD2
LEU
H
11
−3.533
3.937
13.837
1
42.34
H


ATOM
1726
C
LEU
H
11
−5.884
5.725
10.367
1
38.46
H


ATOM
1727
O
LEU
H
11
−4.973
5.866
9.554
1
37.95
H


ATOM
1728
N
MET
H
12
−6.681
6.72
10.742
1
39.26
H


ATOM
1729
CA
MET
H
12
−6.517
8.067
10.208
1
40.3
H


ATOM
1730
CB
MET
H
12
−7.723
8.439
9.338
1
42.96
H


ATOM
1731
CG
MET
H
12
−7.926
7.559
8.111
1
46.74
H


ATOM
1732
SD
MET
H
12
−6.952
8.06
6.679
1
50.76
H


ATOM
1733
CE
MET
H
12
−5.458
7.117
6.92
1
52.67
H


ATOM
1734
C
MET
H
12
−6.373
9.09
11.324
1
40.83
H


ATOM
1735
O
MET
H
12
−6.936
8.922
12.409
1
38.81
H


ATOM
1736
N
LYS
H
13
−5.609
10.146
11.054
1
41.42
H


ATOM
1737
CA
LYS
H
13
−5.409
11.205
12.031
1
41.93
H


ATOM
1738
CB
LYS
H
13
−4.309
12.173
11.576
1
45.64
H


ATOM
1739
CG
LYS
H
13
−2.934
11.546
11.363
1
50.85
H


ATOM
1740
CD
LYS
H
13
−2.304
11.066
12.671
1
53.64
H


ATOM
1741
CE
LYS
H
13
−0.976
10.356
12.411
1
55.04
H


ATOM
1742
NZ
LYS
H
13
−0.323
9.871
13.663
1
57
H


ATOM
1743
C
LYS
H
13
−6.717
11.961
12.1
1
40.27
H


ATOM
1744
O
LYS
H
13
−7.425
12.07
11.102
1
38.48
H


ATOM
1745
N
PRO
H
14
−7.078
12.469
13.282
1
40.78
H


ATOM
1746
CD
PRO
H
14
−6.492
12.295
14.621
1
40.41
H


ATOM
1747
CA
PRO
H
14
−8.337
13.212
13.343
1
41.09
H


ATOM
1748
CB
PRO
H
14
−8.432
13.608
14.814
1
40.82
H


ATOM
1749
CG
PRO
H
14
−7.688
12.5
15.51
1
41.75
H


ATOM
1750
C
PRO
H
14
−8.174
14.428
12.425
1
41.45
H


ATOM
1751
O
PRO
H
14
−7.063
14.948
12.276
1
40.34
H


ATOM
1752
N
GLY
H
15
−9.262
14.866
11.798
1
40.4
H


ATOM
1753
CA
GLY
H
15
−9.181
16.017
10.914
1
38.46
H


ATOM
1754
C
GLY
H
15
−8.88
15.635
9.479
1
38.1
H


ATOM
1755
O
GLY
H
15
−9.232
16.36
8.543
1
39.47
H


ATOM
1756
N
ALA
H
16
−8.226
14.49
9.306
1
36.33
H


ATOM
1757
CA
ALA
H
16
−7.87
13.988
7.984
1
33.38
H


ATOM
1758
CB
ALA
H
16
−6.938
12.79
8.114
1
31.78
H


ATOM
1759
C
ALA
H
16
−9.113
13.593
7.202
1
33.67
H


ATOM
1760
O
ALA
H
16
−10.23
13.612
7.724
1
31.81
H


ATOM
1761
N
SER
H
17
−8.902
13.222
5.945
1
33.93
H


ATOM
1762
CA
SER
H
17
−9.987
12.828
5.062
1
32.25
H


ATOM
1763
CB
SER
H
17
−10.14
13.876
3.96
1
33.54
H


ATOM
1764
OG
SER
H
17
−11.336
13.698
3.228
1
40.78
H


ATOM
1765
C
SER
H
17
−9.651
11.464
4.466
1
31.37
H


ATOM
1766
O
SER
H
17
−8.5
11.196
4.128
1
31.74
H


ATOM
1767
N
VAL
H
18
−10.648
10.597
4.345
1
31.07
H


ATOM
1768
CA
VAL
H
18
−10.413
9.263
3.798
1
29.32
H


ATOM
1769
CB
VAL
H
18
−10.425
8.194
4.902
1
29.78
H


ATOM
1770
CG1
VAL
H
18
−11.861
7.978
5.403
1
24.72
H


ATOM
1771
CG2
VAL
H
18
−9.826
6.903
4.375
1
26.97
H


ATOM
1772
C
VAL
H
18
−11.452
8.857
2.766
1
29.69
H


ATOM
1773
O
VAL
H
18
−12.55
9.409
2.71
1
29.79
H


ATOM
1774
N
GLN
H
19
−11.105
7.867
1.96
1
30.13
H


ATOM
1775
CA
GLN
H
19
−12.021
7.396
0.946
1
31.78
H


ATOM
1776
CB
GLN
H
19
−11.49
7.752
−0.441
1
35.28
H


ATOM
1777
CG
GLN
H
19
−12.442
7.443
−1.573
1
41.49
H


ATOM
1778
CD
GLN
H
19
−11.772
7.499
−2.934
1
45.43
H


ATOM
1779
OE1
GLN
H
19
−10.8
6.78
−3.189
1
48.52
H


ATOM
1780
NE2
GLN
H
19
−12.291
8.347
−3.82
1
44.84
H


ATOM
1781
C
GLN
H
19
−12.18
5.892
1.047
1
31.93
H


ATOM
1782
O
GLN
H
19
−11.205
5.162
0.901
1
33.61
H


ATOM
1783
N
ILE
H
20
−13.398
5.43
1.326
1
29.39
H


ATOM
1784
CA
ILE
H
20
−13.659
3.994
1.368
1
26.84
H


ATOM
1785
CB
ILE
H
20
−14.719
3.591
2.42
1
26.42
H


ATOM
1786
CG2
ILE
H
20
−15.287
2.215
2.079
1
24.44
H


ATOM
1787
CG1
ILE
H
20
−14.098
3.52
3.815
1
29.02
H


ATOM
1788
CD1
ILE
H
20
−13.655
4.838
4.367
1
32.54
H


ATOM
1789
C
ILE
H
20
−14.205
3.62
−0.01
1
24.86
H


ATOM
1790
O
ILE
H
20
−15.044
4.322
−0.569
1
19.67
H


ATOM
1791
N
SER
H
21
−13.718
2.519
−0.562
1
25
H


ATOM
1792
CA
SER
H
21
−14.185
2.086
−1.866
1
27.35
H


ATOM
1793
CB
SER
H
21
−13.008
1.709
−2.766
1
24.52
H


ATOM
1794
OG
SER
H
21
−12.455
0.475
−2.353
1
30.13
H


ATOM
1795
C
SER
H
21
−15.099
0.883
−1.708
1
26.54
H


ATOM
1796
O
SER
H
21
−15.047
0.165
−0.703
1
25.88
H


ATOM
1797
N
CYS
H
22
−15.948
0.685
−2.705
1
25.26
H


ATOM
1798
CA
CYS
H
22
−16.865
−0.435
−2.713
1
26.51
H


ATOM
1799
C
CYS
H
22
−17.05
−0.897
−4.15
1
26.06
H


ATOM
1800
O
CYS
H
22
−17.814
−0.297
−4.916
1
24.2
H


ATOM
1801
CB
CYS
H
22
−18.213
−0.041
−2.125
1
30.22
H


ATOM
1802
SG
CYS
H
22
−19.285
−1.495
−1.9
1
33.16
H


ATOM
1803
N
LYS
H
23
−16.336
−1.961
−4.507
1
25.29
H


ATOM
1804
CA
LYS
H
23
−16.4
−2.519
−5.851
1
27.21
H


ATOM
1805
CB
LYS
H
23
−15.073
−3.193
−6.222
1
30.38
H


ATOM
1806
CG
LYS
H
23
−15.027
−3.666
−7.671
1
35.19
H


ATOM
1807
CD
LYS
H
23
−13.64
−4.122
−8.083
1
37.69
H


ATOM
1808
CE
LYS
H
23
−13.617
−4.582
−9.54
1
36.88
H


ATOM
1809
NZ
LYS
H
23
−14.043
−3.5
−10.468
1
35.83
H


ATOM
1810
C
LYS
H
23
−17.531
−3.527
−5.983
1
24.02
H


ATOM
1811
O
LYS
H
23
−17.544
−4.552
−5.313
1
22.08
H


ATOM
1812
N
ALA
H
24
−18.476
−3.221
−6.862
1
25.47
H


ATOM
1813
CA
ALA
H
24
−19.627
−4.084
−7.095
1
24.88
H


ATOM
1814
CB
ALA
H
24
−20.881
−3.235
−7.253
1
20.38
H


ATOM
1815
C
ALA
H
24
−19.446
−4.965
−8.328
1
26.09
H


ATOM
1816
O
ALA
H
24
−18.975
−4.515
−9.378
1
26.88
H


ATOM
1817
N
THR
H
25
−19.81
−6.231
−8.191
1
24.75
H


ATOM
1818
CA
THR
H
25
−19.73
−7.156
−9.304
1
23.77
H


ATOM
1819
CB
THR
H
25
−18.397
−7.921
−9.315
1
22.97
H


ATOM
1820
OG1
THR
H
25
−18.238
−8.626
−8.081
1
23.52
H


ATOM
1821
CG2
THR
H
25
−17.239
−6.962
−9.505
1
21.95
H


ATOM
1822
C
THR
H
25
−20.877
−8.138
−9.154
1
23.54
H


ATOM
1823
O
THR
H
25
−21.458
−8.263
−8.076
1
21.65
H


ATOM
1824
N
GLY
H
26
−21.218
−8.819
−10.24
1
23.86
H


ATOM
1825
CA
GLY
H
26
−22.294
−9.792
−10.181
1
24.4
H


ATOM
1826
C
GLY
H
26
−23.692
−9.253
−10.44
1
24.22
H


ATOM
1827
O
GLY
H
26
−24.669
−9.997
−10.324
1
23.31
H


ATOM
1828
N
TYR
H
27
−23.797
−7.971
−10.786
1
21.92
H


ATOM
1829
CA
TYR
H
27
−25.097
−7.369
−11.064
1
21.3
H


ATOM
1830
CB
TYR
H
27
−25.862
−7.149
−9.753
1
22.54
H


ATOM
1831
CG
TYR
H
27
−25.364
−5.986
−8.929
1
20.9
H


ATOM
1832
CD1
TYR
H
27
−25.857
−4.696
−9.138
1
18.51
H


ATOM
1833
CE1
TYR
H
27
−25.414
−3.621
−8.375
1
16.6
H


ATOM
1834
CD2
TYR
H
27
−24.405
−6.173
−7.935
1
22.7
H


ATOM
1835
CE2
TYR
H
27
−23.95
−5.101
−7.165
1
21.37
H


ATOM
1836
CZ
TYR
H
27
−24.464
−3.831
−7.388
1
19.5
H


ATOM
1837
OH
TYR
H
27
−24.047
−2.785
−6.599
1
18.29
H


ATOM
1838
C
TYR
H
27
−24.948
−6.044
−11.8
1
19.83
H


ATOM
1839
O
TYR
H
27
−23.852
−5.491
−11.883
1
19.39
H


ATOM
1840
N
THR
H
28
−26.057
−5.536
−12.329
1
19.67
H


ATOM
1841
CA
THR
H
28
−26.049
−4.266
−13.047
1
17.55
H


ATOM
1842
CB
THR
H
28
−27.348
−4.073
−13.82
1
17.4
H


ATOM
1843
OG1
THR
H
28
−27.634
−5.271
−14.56
1
20.26
H


ATOM
1844
CG2
THR
H
28
−27.222
−2.898
−14.787
1
14.93
H


ATOM
1845
C
THR
H
28
−25.86
−3.117
−12.061
1
19.8
H


ATOM
1846
O
THR
H
28
−26.816
−2.619
−11.45
1
19.37
H


ATOM
1847
N
PHE
H
29
−24.6
−2.719
−11.914
1
21.18
H


ATOM
1848
CA
PHE
H
29
−24.18
−1.657
−11.009
1
22.53
H


ATOM
1849
CB
PHE
H
29
−22.772
−1.18
−11.396
1
21.17
H


ATOM
1850
CG
PHE
H
29
−22.197
−0.141
−10.469
1
18.49
H


ATOM
1851
CD1
PHE
H
29
−21.855
−0.466
−9.161
1
19.16
H


ATOM
1852
CD2
PHE
H
29
−21.993
1.162
−10.907
1
19.6
H


ATOM
1853
CE1
PHE
H
29
−21.316
0.493
−8.3
1
18.58
H


ATOM
1854
CE2
PHE
H
29
−21.452
2.132
−10.051
1
21.63
H


ATOM
1855
CZ
PHE
H
29
−21.115
1.792
−8.743
1
19.01
H


ATOM
1856
C
PHE
H
29
−25.121
−0.462
−10.98
1
24.39
H


ATOM
1857
O
PHE
H
29
−25.622
−0.085
−9.925
1
24.92
H


ATOM
1858
N
SER
H
30
−25.361
0.121
−12.149
1
24.35
H


ATOM
1859
CA
SER
H
30
−26.197
1.307
−12.265
1
24.36
H


ATOM
1860
CB
SER
H
30
−26.094
1.864
−13.685
1
22.4
H


ATOM
1861
OG
SER
H
30
−26.714
0.985
−14.607
1
24.52
H


ATOM
1862
C
SER
H
30
−27.678
1.194
−11.887
1
25.6
H


ATOM
1863
O
SER
H
30
−28.312
2.216
−11.62
1
26.55
H


ATOM
1864
N
ASP
H
31
−28.246
−0.009
−11.85
1
24.12
H


ATOM
1865
CA
ASP
H
31
−29.672
−0.104
−11.518
1
23.11
H


ATOM
1866
CB
ASP
H
31
−30.311
−1.354
−12.126
1
20.9
H


ATOM
1867
CG
ASP
H
31
−30.311
−1.343
−13.64
1
21.07
H


ATOM
1868
OD1
ASP
H
31
−30.177
−0.26
−14.251
1
21.33
H


ATOM
1869
OD2
ASP
H
31
−30.464
−2.432
−14.221
1
21.23
H


ATOM
1870
C
ASP
H
31
−29.985
−0.102
−10.034
1
24.52
H


ATOM
1871
O
ASP
H
31
−31.148
−0.233
−9.648
1
26.44
H


ATOM
1872
N
TYR
H
32
−28.97
0.057
−9.195
1
23.91
H


ATOM
1873
CA
TYR
H
32
−29.218
0.024
−7.766
1
21.89
H


ATOM
1874
CB
TYR
H
32
−28.696
−1.283
−7.17
1
22.5
H


ATOM
1875
CG
TYR
H
32
−29.387
−2.527
−7.674
1
22.19
H


ATOM
1876
CD1
TYR
H
32
−29.099
−3.046
−8.933
1
23.07
H


ATOM
1877
CE1
TYR
H
32
−29.722
−4.205
−9.396
1
23.2
H


ATOM
1878
CD2
TYR
H
32
−30.323
−3.193
−6.885
1
21.22
H


ATOM
1879
CE2
TYR
H
32
−30.952
−4.347
−7.337
1
20.23
H


ATOM
1880
CZ
TYR
H
32
−30.644
−4.849
−8.594
1
22.35
H


ATOM
1881
OH
TYR
H
32
−31.255
−5.997
−9.048
1
22.85
H


ATOM
1882
C
TYR
H
32
−28.655
1.166
−6.955
1
23.86
H


ATOM
1883
O
TYR
H
32
−27.876
1.99
−7.436
1
23.76
H


ATOM
1884
N
TRP
H
33
−29.077
1.195
−5.698
1
22.85
H


ATOM
1885
CA
TRP
H
33
−28.623
2.19
−4.753
1
22.55
H


ATOM
1886
CB
TRP
H
33
−29.77
2.686
−3.869
1
21.08
H


ATOM
1887
CG
TRP
H
33
−30.734
3.631
−4.506
1
20.86
H


ATOM
1888
CD2
TRP
H
33
−30.787
5.045
−4.307
1
20.3
H


ATOM
1889
CE2
TRP
H
33
−31.902
5.529
−5.035
1
19.41
H


ATOM
1890
CE3
TRP
H
33
−30.003
5.953
−3.581
1
20.24
H


ATOM
1891
CD1
TRP
H
33
−31.782
3.319
−5.332
1
19.75
H


ATOM
1892
NE1
TRP
H
33
−32.49
4.455
−5.649
1
16.56
H


ATOM
1893
CZ2
TRP
H
33
−32.254
6.885
−5.056
1
20.07
H


ATOM
1894
CZ3
TRP
H
33
−30.354
7.307
−3.601
1
23.51
H


ATOM
1895
CH2
TRP
H
33
−31.472
7.757
−4.335
1
20.45
H


ATOM
1896
C
TRP
H
33
−27.635
1.478
−3.854
1
23.23
H


ATOM
1897
O
TRP
H
33
−27.898
0.361
−3.406
1
23.43
H


ATOM
1898
N
ILE
H
34
−26.497
2.11
−3.601
1
21.7
H


ATOM
1899
CA
ILE
H
34
−25.519
1.541
−2.687
1
19.36
H


ATOM
1900
CB
ILE
H
34
−24.066
1.743
−3.187
1
19.9
H


ATOM
1901
CG2
ILE
H
34
−23.086
1.483
−2.056
1
19.58
H


ATOM
1902
CG1
ILE
H
34
−23.774
0.797
−4.357
1
18.89
H


ATOM
1903
CD1
ILE
H
34
−23.809
−0.669
−3.979
1
21.63
H


ATOM
1904
C
ILE
H
34
−25.736
2.309
−1.384
1
18.33
H


ATOM
1905
O
ILE
H
34
−25.782
3.542
−1.379
1
16.4
H


ATOM
1906
N
GLU
H
35
−25.905
1.58
−0.287
1
18.42
H


ATOM
1907
CA
GLU
H
35
−26.121
2.205
1.01
1
18.46
H


ATOM
1908
CB
GLU
H
35
−27.126
1.412
1.851
1
18.97
H


ATOM
1909
CG
GLU
H
35
−28.435
1.135
1.17
1
21.62
H


ATOM
1910
CD
GLU
H
35
−29.15
2.396
0.78
1
19.89
H


ATOM
1911
OE1
GLU
H
35
−29.586
3.135
1.684
1
20.57
H


ATOM
1912
OE2
GLU
H
35
−29.269
2.648
−0.435
1
26.49
H


ATOM
1913
C
GLU
H
35
−24.817
2.23
1.766
1
19.64
H


ATOM
1914
O
GLU
H
35
−23.964
1.365
1.575
1
20.3
H


ATOM
1915
N
TRP
H
36
−24.669
3.227
2.626
1
18.16
H


ATOM
1916
CA
TRP
H
36
−23.495
3.333
3.454
1
18.95
H


ATOM
1917
CB
TRP
H
36
−22.718
4.608
3.13
1
18.6
H


ATOM
1918
CG
TRP
H
36
−22.005
4.51
1.814
1
21.47
H


ATOM
1919
CD2
TRP
H
36
−20.77
3.832
1.563
1
18.88
H


ATOM
1920
CE2
TRP
H
36
−20.5
3.953
0.185
1
21.86
H


ATOM
1921
CE3
TRP
H
36
−19.866
3.131
2.371
1
20.92
H


ATOM
1922
CD1
TRP
H
36
−22.426
5.001
0.606
1
21.64
H


ATOM
1923
NE1
TRP
H
36
−21.526
4.67
−0.377
1
22.42
H


ATOM
1924
CZ2
TRP
H
36
−19.354
3.398
−0.407
1
24.84
H


ATOM
1925
CZ3
TRP
H
36
−18.729
2.578
1.785
1
22.34
H


ATOM
1926
CH2
TRP
H
36
−18.484
2.716
0.408
1
22.84
H


ATOM
1927
C
TRP
H
36
−24.001
3.329
4.888
1
22.08
H


ATOM
1928
O
TRP
H
36
−24.678
4.261
5.331
1
22.48
H


ATOM
1929
N
VAL
H
37
−23.695
2.255
5.605
1
23.94
H


ATOM
1930
CA
VAL
H
37
−24.143
2.127
6.979
1
26.78
H


ATOM
1931
CB
VAL
H
37
−24.917
0.814
7.204
1
27.47
H


ATOM
1932
CG1
VAL
H
37
−25.416
0.746
8.65
1
24.44
H


ATOM
1933
CG2
VAL
H
37
−26.084
0.729
6.235
1
26.91
H


ATOM
1934
C
VAL
H
37
−22.99
2.172
7.956
1
28.44
H


ATOM
1935
O
VAL
H
37
−21.958
1.52
7.764
1
26.59
H


ATOM
1936
N
LYS
H
38
−23.186
2.939
9.018
1
28.56
H


ATOM
1937
CA
LYS
H
38
−22.167
3.087
10.034
1
30.4
H


ATOM
1938
CB
LYS
H
38
−22.016
4.572
10.376
1
32.38
H


ATOM
1939
CG
LYS
H
38
−21.131
4.892
11.563
1
31.19
H


ATOM
1940
CD
LYS
H
38
−21.092
6.39
11.783
1
33.36
H


ATOM
1941
CE
LYS
H
38
−20.284
6.748
13.015
1
39.61
H


ATOM
1942
NZ
LYS
H
38
−20.029
8.21
13.122
1
38.99
H


ATOM
1943
C
LYS
H
38
−22.535
2.297
11.28
1
31.25
H


ATOM
1944
O
LYS
H
38
−23.713
2.107
11.581
1
27.61
H


ATOM
1945
N
GLN
H
39
−21.515
1.818
11.983
1
32.31
H


ATOM
1946
CA
GLN
H
39
−21.714
1.092
13.227
1
34.45
H


ATOM
1947
CB
GLN
H
39
−21.982
−0.396
12.99
1
35.26
H


ATOM
1948
CG
GLN
H
39
−22.102
−1.191
14.304
1
37.54
H


ATOM
1949
CD
GLN
H
39
−22.299
−2.685
14.096
1
37.74
H


ATOM
1950
OE1
GLN
H
39
−21.676
−3.292
13.219
1
39.37
H


ATOM
1951
NE2
GLN
H
39
−23.152
−3.288
14.918
1
36.99
H


ATOM
1952
C
GLN
H
39
−20.497
1.247
14.126
1
35.83
H


ATOM
1953
O
GLN
H
39
−19.479
0.577
13.938
1
31.33
H


ATOM
1954
N
ARG
H
40
−20.602
2.161
15.085
1
40.48
H


ATOM
1955
CA
ARG
H
40
−19.531
2.384
16.044
1
44.61
H


ATOM
1956
CB
ARG
H
40
−19.805
3.64
16.867
1
46
H


ATOM
1957
CG
ARG
H
40
−19.921
4.929
16.054
1
47.23
H


ATOM
1958
CD
ARG
H
40
−20.17
6.121
16.971
1
51.07
H


ATOM
1959
NE
ARG
H
40
−19.057
6.348
17.895
1
53.99
H


ATOM
1960
CZ
ARG
H
40
−19.109
7.17
18.941
1
56.17
H


ATOM
1961
NH1
ARG
H
40
−20.223
7.844
19.205
1
56.52
H


ATOM
1962
NH2
ARG
H
40
−18.044
7.33
19.718
1
55.66
H


ATOM
1963
C
ARG
H
40
−19.544
1.142
16.934
1
47.66
H


ATOM
1964
O
ARG
H
40
−20.608
0.699
17.376
1
47.1
H


ATOM
1965
N
PRO
H
41
−18.36
0.566
17.203
1
50.56
H


ATOM
1966
CD
PRO
H
41
−17.081
1.273
16.999
1
51.94
H


ATOM
1967
CA
PRO
H
41
−18.15
−0.637
18.019
1
52.23
H


ATOM
1968
CB
PRO
H
41
−16.798
−0.37
18.663
1
53.82
H


ATOM
1969
CG
PRO
H
41
−16.058
0.271
17.53
1
54.36
H


ATOM
1970
C
PRO
H
41
−19.225
−1.034
19.032
1
52.28
H


ATOM
1971
O
PRO
H
41
−19.808
−2.115
18.935
1
52.75
H


ATOM
1972
N
GLY
H
42
−19.489
−0.179
20.009
1
51.81
H


ATOM
1973
CA
GLY
H
42
−20.491
−0.537
20.992
1
55.15
H


ATOM
1974
C
GLY
H
42
−21.912
−0.169
20.61
1
56.46
H


ATOM
1975
O
GLY
H
42
−22.864
−0.548
21.296
1
55.69
H


ATOM
1976
N
HIS
H
43
−22.059
0.546
19.5
1
57.72
H


ATOM
1977
CA
HIS
H
43
−23.369
0.999
19.046
1
57.08
H


ATOM
1978
CB
HIS
H
43
−23.218
2.386
18.426
1
58.6
H


ATOM
1979
CG
HIS
H
43
−22.676
3.404
19.38
1
62.89
H


ATOM
1980
CD2
HIS
H
43
−21.414
3.85
19.587
1
64.88
H


ATOM
1981
ND1
HIS
H
43
−23.47
4.063
20.294
1
66.25
H


ATOM
1982
CE1
HIS
H
43
−22.721
4.875
21.02
1
67.36
H


ATOM
1983
NE2
HIS
H
43
−21.469
4.764
20.612
1
67.26
H


ATOM
1984
C
HIS
H
43
−24.093
0.069
18.078
1
55.71
H


ATOM
1985
O
HIS
H
43
−23.619
−1.029
17.766
1
54.99
H


ATOM
1986
N
GLY
H
44
−25.26
0.52
17.622
1
52.16
H


ATOM
1987
CA
GLY
H
44
−26.044
−0.255
16.68
1
48.21
H


ATOM
1988
C
GLY
H
44
−25.774
0.217
15.262
1
45.07
H


ATOM
1989
O
GLY
H
44
−24.668
0.659
14.941
1
44.58
H


ATOM
1990
N
LEU
H
45
−26.788
0.128
14.41
1
41.56
H


ATOM
1991
CA
LEU
H
45
−26.661
0.538
13.018
1
37.28
H


ATOM
1992
CB
LEU
H
45
−27.291
−0.515
12.108
1
36.2
H


ATOM
1993
CG
LEU
H
45
−26.493
−1.752
11.694
1
35.94
H


ATOM
1994
CD1
LEU
H
45
−25.679
−2.29
12.856
1
36.28
H


ATOM
1995
CD2
LEU
H
45
−27.468
−2.797
11.172
1
31.97
H


ATOM
1996
C
LEU
H
45
−27.304
1.884
12.707
1
36.54
H


ATOM
1997
O
LEU
H
45
−28.451
2.139
13.073
1
38.5
H


ATOM
1998
N
GLU
H
46
−26.561
2.743
12.021
1
35.04
H


ATOM
1999
CA
GLU
H
46
−27.093
4.033
11.618
1
35.12
H


ATOM
2000
CB
GLU
H
46
−26.563
5.158
12.517
1
38.57
H


ATOM
2001
CG
GLU
H
46
−25.077
5.211
12.695
1
46.18
H


ATOM
2002
CD
GLU
H
46
−24.669
6.258
13.72
1
51.8
H


ATOM
2003
OE1
GLU
H
46
−25.111
7.424
13.595
1
54.76
H


ATOM
2004
OE2
GLU
H
46
−23.903
5.914
14.647
1
54.83
H


ATOM
2005
C
GLU
H
46
−26.812
4.318
10.14
1
30.78
H


ATOM
2006
O
GLU
H
46
−25.707
4.11
9.635
1
30.57
H


ATOM
2007
N
TRP
H
47
−27.849
4.776
9.456
1
26.63
H


ATOM
2008
CA
TRP
H
47
−27.796
5.086
8.034
1
26.33
H


ATOM
2009
CB
TRP
H
47
−29.224
5.227
7.508
1
24.4
H


ATOM
2010
CG
TRP
H
47
−29.334
5.42
6.027
1
25.57
H


ATOM
2011
CD2
TRP
H
47
−29.812
6.59
5.354
1
22.44
H


ATOM
2012
CE2
TRP
H
47
−29.844
6.295
3.974
1
22.06
H


ATOM
2013
CE3
TRP
H
47
−30.221
7.858
5.786
1
23.3
H


ATOM
2014
CD1
TRP
H
47
−29.09
4.49
5.053
1
24.8
H


ATOM
2015
NE1
TRP
H
47
−29.398
5.008
3.817
1
22.98
H


ATOM
2016
CZ2
TRP
H
47
−30.274
7.223
3.019
1
23.86
H


ATOM
2017
CZ3
TRP
H
47
−30.649
8.783
4.84
1
25.22
H


ATOM
2018
CH2
TRP
H
47
−30.672
8.459
3.469
1
25.87
H


ATOM
2019
C
TRP
H
47
−27.011
6.355
7.716
1
23.65
H


ATOM
2020
O
TRP
H
47
−27.354
7.438
8.18
1
24.01
H


ATOM
2021
N
ILE
H
48
−25.965
6.232
6.912
1
20.54
H


ATOM
2022
CA
ILE
H
48
−25.18
7.407
6.558
1
19.71
H


ATOM
2023
CB
ILE
H
48
−23.723
7.028
6.279
1
17.95
H


ATOM
2024
CG2
ILE
H
48
−22.931
8.27
5.884
1
18.96
H


ATOM
2025
CG1
ILE
H
48
−23.122
6.369
7.524
1
17.37
H


ATOM
2026
CD1
ILE
H
48
−21.716
5.859
7.332
1
16.67
H


ATOM
2027
C
ILE
H
48
−25.762
8.099
5.323
1
20.71
H


ATOM
2028
O
ILE
H
48
−25.717
9.323
5.191
1
19.38
H


ATOM
2029
N
GLY
H
49
−26.317
7.294
4.425
1
22.82
H


ATOM
2030
CA
GLY
H
49
−26.898
7.813
3.204
1
19.83
H


ATOM
2031
C
GLY
H
49
−26.678
6.803
2.101
1
19.96
H


ATOM
2032
O
GLY
H
49
−26.064
5.76
2.323
1
20.94
H


ATOM
2033
N
ASP
H
50
−27.185
7.099
0.912
1
18.66
H


ATOM
2034
CA
ASP
H
50
−27.017
6.199
−0.214
1
17.79
H


ATOM
2035
CB
ASP
H
50
−28.218
5.255
−0.352
1
16.18
H


ATOM
2036
CG
ASP
H
50
−29.564
5.982
−0.299
1
22.26
H


ATOM
2037
OD1
ASP
H
50
−29.644
7.197
−0.605
1
21.47
H


ATOM
2038
OD2
ASP
H
50
−30.562
5.319
0.045
1
23.16
H


ATOM
2039
C
ASP
H
50
−26.828
6.991
−1.494
1
18.09
H


ATOM
2040
O
ASP
H
50
−26.895
8.219
−1.492
1
19.7
H


ATOM
2041
N
ILE
H
51
−26.588
6.278
−2.586
1
19.3
H


ATOM
2042
CA
ILE
H
51
−26.387
6.909
−3.878
1
19.6
H


ATOM
2043
CB
ILE
H
51
−24.899
7.23
−4.092
1
20.23
H


ATOM
2044
CG2
ILE
H
51
−24.057
5.97
−3.888
1
17.18
H


ATOM
2045
CG1
ILE
H
51
−24.692
7.86
−5.47
1
19.45
H


ATOM
2046
CD1
ILE
H
51
−23.294
8.421
−5.669
1
18.02
H


ATOM
2047
C
ILE
H
51
−26.878
6.002
−4.997
1
21.67
H


ATOM
2048
O
ILE
H
51
−26.547
4.816
−5.042
1
23.17
H


ATOM
2049
N
LEU
H
52
−27.689
6.559
−5.889
1
23.52
H


ATOM
2050
CA
LEU
H
52
−28.214
5.796
−7.015
1
22.8
H


ATOM
2051
CB
LEU
H
52
−29.472
6.463
−7.572
1
19.93
H


ATOM
2052
CG
LEU
H
52
−30.56
5.528
−8.11
1
19.45
H


ATOM
2053
CD1
LEU
H
52
−31.483
6.341
−9.01
1
17
H


ATOM
2054
CD2
LEU
H
52
−29.952
4.354
−8.868
1
10.59
H


ATOM
2055
C
LEU
H
52
−27.113
5.762
−8.079
1
22.63
H


ATOM
2056
O
LEU
H
52
−27.002
6.659
−8.912
1
22.12
H


ATOM
2057
N
CYS
H
52A
−26.304
4.712
−8.03
1
22.25
H


ATOM
2058
CA
CYS
H
52A
−25.186
4.544
−8.939
1
24.62
H


ATOM
2059
CB
CYS
H
52A
−24.647
3.12
−8.804
1
24.23
H


ATOM
2060
SG
CYS
H
52A
−24.071
2.736
−7.113
1
22.95
H


ATOM
2061
C
CYS
H
52A
−25.479
4.874
−10.4
1
26.06
H


ATOM
2062
O
CYS
H
52A
−24.602
5.342
−11.128
1
28.55
H


ATOM
2063
N
GLY
H
53
−26.714
4.652
−10.828
1
24.42
H


ATOM
2064
CA
GLY
H
53
−27.061
4.927
−12.207
1
23.12
H


ATOM
2065
C
GLY
H
53
−27.24
6.389
−12.567
1
24.51
H


ATOM
2066
O
GLY
H
53
−27.046
6.763
−13.724
1
27.27
H


ATOM
2067
N
THR
H
54
−27.611
7.224
−11.602
1
22.58
H


ATOM
2068
CA
THR
H
54
−27.815
8.636
−11.9
1
23.15
H


ATOM
2069
CB
THR
H
54
−29.263
9.064
−11.636
1
24.4
H


ATOM
2070
OG1
THR
H
54
−29.489
9.128
−10.223
1
21.63
H


ATOM
2071
CG2
THR
H
54
−30.228
8.074
−12.261
1
22.77
H


ATOM
2072
C
THR
H
54
−26.921
9.548
−11.083
1
23.15
H


ATOM
2073
O
THR
H
54
−26.739
10.716
−11.422
1
26.47
H


ATOM
2074
N
GLY
H
55
−26.381
9.02
−9.995
1
21.87
H


ATOM
2075
CA
GLY
H
55
−25.522
9.82
−9.154
1
18.75
H


ATOM
2076
C
GLY
H
55
−26.298
10.539
−8.074
1
20.96
H


ATOM
2077
O
GLY
H
55
−25.712
11.277
−7.284
1
23.14
H


ATOM
2078
N
ARG
H
56
−27.613
10.331
−8.02
1
21.63
H


ATOM
2079
CA
ARG
H
56
−28.415
10.996
−6.999
1
21.59
H


ATOM
2080
CB
ARG
H
56
−29.904
10.683
−7.165
1
21.22
H


ATOM
2081
CG
ARG
H
56
−30.764
11.464
−6.181
1
20.85
H


ATOM
2082
CD
ARG
H
56
−32.199
10.971
−6.117
1
18.23
H


ATOM
2083
NE
ARG
H
56
−32.89
11.535
−4.956
1
15.79
H


ATOM
2084
CZ
ARG
H
56
−34.129
11.223
−4.594
1
16.38
H


ATOM
2085
NH1
ARG
H
56
−34.824
10.351
−5.307
1
14.78
H


ATOM
2086
NH2
ARG
H
56
−34.67
11.774
−3.511
1
17.33
H


ATOM
2087
C
ARG
H
56
−27.959
10.556
−5.612
1
21.59
H


ATOM
2088
O
ARG
H
56
−27.71
9.378
−5.371
1
22.43
H


ATOM
2089
N
THR
H
57
−27.852
11.508
−4.697
1
22.54
H


ATOM
2090
CA
THR
H
57
−27.417
11.19
−3.345
1
23.23
H


ATOM
2091
CB
THR
H
57
−26.048
11.839
−3.009
1
21.89
H


ATOM
2092
OG1
THR
H
57
−26.145
13.255
−3.161
1
20.5
H


ATOM
2093
CG2
THR
H
57
−24.949
11.31
−3.921
1
20.02
H


ATOM
2094
C
THR
H
57
−28.407
11.657
−2.299
1
23.21
H


ATOM
2095
O
THR
H
57
−29.097
12.66
−2.476
1
25.08
H


ATOM
2096
N
ARG
H
58
−28.477
10.902
−1.212
1
22.91
H


ATOM
2097
CA
ARG
H
58
−29.338
11.225
−0.091
1
21.69
H


ATOM
2098
CB
ARG
H
58
−30.564
10.307
−0.049
1
21.46
H


ATOM
2099
CG
ARG
H
58
−31.557
10.528
−1.174
1
18.91
H


ATOM
2100
CD
ARG
H
58
−32.816
9.708
−0.973
1
16.44
H


ATOM
2101
NE
ARG
H
58
−32.529
8.277
−0.945
1
18.57
H


ATOM
2102
CZ
ARG
H
58
−33.441
7.331
−0.73
1
20.56
H


ATOM
2103
NH1
ARG
H
58
−34.716
7.661
−0.52
1
17.73
H


ATOM
2104
NH2
ARG
H
58
−33.078
6.051
−0.725
1
16.01
H


ATOM
2105
C
ARG
H
58
−28.478
11.016
1.146
1
23.65
H


ATOM
2106
O
ARG
H
58
−27.971
9.92
1.389
1
24.01
H


ATOM
2107
N
TYR
H
59
−28.304
12.081
1.917
1
26.46
H


ATOM
2108
CA
TYR
H
59
−27.485
12.03
3.119
1
28.75
H


ATOM
2109
CB
TYR
H
59
−26.444
13.147
3.082
1
24.76
H


ATOM
2110
CG
TYR
H
59
−25.493
13.067
1.923
1
23.56
H


ATOM
2111
CD1
TYR
H
59
−24.463
12.141
1.917
1
21.53
H


ATOM
2112
CE1
TYR
H
59
−23.586
12.049
0.849
1
22.58
H


ATOM
2113
CD2
TYR
H
59
−25.631
13.911
0.823
1
22.39
H


ATOM
2114
CE2
TYR
H
59
−24.759
13.828
−0.257
1
23.74
H


ATOM
2115
CZ
TYR
H
59
−23.735
12.891
−0.234
1
23.29
H


ATOM
2116
OH
TYR
H
59
−22.85
12.789
−1.288
1
26.35
H


ATOM
2117
C
TYR
H
59
−28.286
12.177
4.405
1
31.04
H


ATOM
2118
O
TYR
H
59
−29.295
12.887
4.445
1
30.15
H


ATOM
2119
N
ASN
H
60
−27.83
11.495
5.453
1
31.95
H


ATOM
2120
CA
ASN
H
60
−28.465
11.614
6.751
1
34.28
H


ATOM
2121
CB
ASN
H
60
−28.025
10.484
7.686
1
32.08
H


ATOM
2122
CG
ASN
H
60
−28.923
10.353
8.907
1
32.62
H


ATOM
2123
OD1
ASN
H
60
−29.277
11.347
9.546
1
32.55
H


ATOM
2124
ND2
ASN
H
60
−29.289
9.119
9.242
1
32.09
H


ATOM
2125
C
ASN
H
60
−27.898
12.953
7.227
1
36.98
H


ATOM
2126
O
ASN
H
60
−26.914
13.008
7.971
1
35.09
H


ATOM
2127
N
GLU
H
61
−28.519
14.026
6.748
1
41.41
H


ATOM
2128
CA
GLU
H
61
−28.129
15.402
7.056
1
46.47
H


ATOM
2129
CB
GLU
H
61
−29.296
16.344
6.72
1
50.66
H


ATOM
2130
CG
GLU
H
61
−29.788
16.276
5.27
1
57.21
H


ATOM
2131
CD
GLU
H
61
−28.826
16.927
4.28
1
60.6
H


ATOM
2132
OE1
GLU
H
61
−28.482
18.112
4.481
1
63.78
H


ATOM
2133
OE2
GLU
H
61
−28.419
16.262
3.3
1
61.22
H


ATOM
2134
C
GLU
H
61
−27.684
15.664
8.5
1
46.04
H


ATOM
2135
O
GLU
H
61
−26.693
16.36
8.736
1
44.28
H


ATOM
2136
N
LYS
H
62
−28.418
15.095
9.452
1
46.41
H


ATOM
2137
CA
LYS
H
62
−28.154
15.289
10.873
1
47.28
H


ATOM
2138
CB
LYS
H
62
−29.355
14.798
11.686
1
46.97
H


ATOM
2139
CG
LYS
H
62
−29.31
13.325
12.055
1
49.13
H


ATOM
2140
CD
LYS
H
62
−30.493
12.959
12.935
1
50.46
H


ATOM
2141
CE
LYS
H
62
−30.232
11.681
13.707
1
53.67
H


ATOM
2142
NZ
LYS
H
62
−29.048
11.82
14.603
1
55.56
H


ATOM
2143
C
LYS
H
62
−26.883
14.681
11.466
1
46.68
H


ATOM
2144
O
LYS
H
62
−26.538
14.97
12.61
1
47.76
H


ATOM
2145
N
LEU
H
63
−26.179
13.846
10.719
1
46.97
H


ATOM
2146
CA
LEU
H
63
−24.98
13.245
11.283
1
47.83
H


ATOM
2147
CB
LEU
H
63
−24.631
11.952
10.546
1
45.64
H


ATOM
2148
CG
LEU
H
63
−25.234
10.7
11.194
1
44.86
H


ATOM
2149
CD1
LEU
H
63
−26.643
10.991
11.677
1
45.16
H


ATOM
2150
CD2
LEU
H
63
−25.217
9.545
10.202
1
44.38
H


ATOM
2151
C
LEU
H
63
−23.778
14.163
11.334
1
49.14
H


ATOM
2152
O
LEU
H
63
−23.44
14.836
10.357
1
46.89
H


ATOM
2153
N
LYS
H
64
−23.15
14.188
12.507
1
52.76
H


ATOM
2154
CA
LYS
H
64
−21.966
14.998
12.747
1
55.38
H


ATOM
2155
CB
LYS
H
64
−21.658
15.067
14.244
1
57.52
H


ATOM
2156
CG
LYS
H
64
−22.678
15.827
15.075
1
60.99
H


ATOM
2157
CD
LYS
H
64
−22.216
15.908
16.523
1
63.36
H


ATOM
2158
CE
LYS
H
64
−23.105
16.811
17.356
1
63.31
H


ATOM
2159
NZ
LYS
H
64
−22.64
16.848
18.771
1
65.48
H


ATOM
2160
C
LYS
H
64
−20.794
14.362
12.018
1
55.7
H


ATOM
2161
O
LYS
H
64
−19.9
13.782
12.638
1
57.16
H


ATOM
2162
N
ALA
H
65
−20.82
14.465
10.693
1
55.2
H


ATOM
2163
CA
ALA
H
65
−19.777
13.914
9.836
1
53.24
H


ATOM
2164
CB
ALA
H
65
−19.797
12.384
9.875
1
53.25
H


ATOM
2165
C
ALA
H
65
−20.033
14.406
8.423
1
51.43
H


ATOM
2166
O
ALA
H
65
−21.184
14.549
7.998
1
50.07
H


ATOM
2167
N
MET
H
66
−18.956
14.665
7.699
1
47.89
H


ATOM
2168
CA
MET
H
66
−19.076
15.159
6.346
1
46.16
H


ATOM
2169
CB
MET
H
66
−18.089
16.3
6.156
1
49.61
H


ATOM
2170
CG
MET
H
66
−18.035
17.199
7.381
1
54.5
H


ATOM
2171
SD
MET
H
66
−17.455
18.875
7.071
1
63.85
H


ATOM
2172
CE
MET
H
66
−19.064
19.742
6.86
1
59.59
H


ATOM
2173
C
MET
H
66
−18.832
14.03
5.354
1
43.02
H


ATOM
2174
O
MET
H
66
−17.696
13.62
5.13
1
41.87
H


ATOM
2175
N
ALA
H
67
−19.921
13.529
4.773
1
39.48
H


ATOM
2176
CA
ALA
H
67
−19.868
12.434
3.814
1
36.35
H


ATOM
2177
CB
ALA
H
67
−20.869
11.371
4.199
1
33.12
H


ATOM
2178
C
ALA
H
67
−20.122
12.87
2.378
1
36.2
H


ATOM
2179
O
ALA
H
67
−20.839
13.837
2.111
1
35.86
H


ATOM
2180
N
THR
H
68
−19.534
12.126
1.453
1
35.87
H


ATOM
2181
CA
THR
H
68
−19.674
12.398
0.033
1
35.01
H


ATOM
2182
CB
THR
H
68
−18.513
13.274
−0.483
1
35.44
H


ATOM
2183
OG1
THR
H
68
−18.578
14.564
0.132
1
35.02
H


ATOM
2184
CG2
THR
H
68
−18.589
13.43
−1.993
1
37.2
H


ATOM
2185
C
THR
H
68
−19.665
11.071
−0.716
1
34.17
H


ATOM
2186
O
THR
H
68
−18.734
10.273
−0.575
1
33.07
H


ATOM
2187
N
PHE
H
69
−20.707
10.834
−1.503
1
31.6
H


ATOM
2188
CA
PHE
H
69
−20.798
9.603
−2.265
1
28.86
H


ATOM
2189
CB
PHE
H
69
−22.185
8.964
−2.112
1
26.83
H


ATOM
2190
CG
PHE
H
69
−22.622
8.78
−0.687
1
27.85
H


ATOM
2191
CD1
PHE
H
69
−21.726
8.334
0.283
1
27.07
H


ATOM
2192
CD2
PHE
H
69
−23.939
9.036
−0.315
1
27.53
H


ATOM
2193
CE1
PHE
H
69
−22.135
8.146
1.603
1
27.42
H


ATOM
2194
CE2
PHE
H
69
−24.364
8.853
1.002
1
26.6
H


ATOM
2195
CZ
PHE
H
69
−23.46
8.407
1.965
1
28.96
H


ATOM
2196
C
PHE
H
69
−20.555
9.882
−3.736
1
28.68
H


ATOM
2197
O
PHE
H
69
−20.999
10.895
−4.264
1
28.94
H


ATOM
2198
N
THR
H
70
−19.834
8.98
−4.387
1
26.97
H


ATOM
2199
CA
THR
H
70
−19.573
9.079
−5.811
1
27.96
H


ATOM
2200
CB
THR
H
70
−18.175
9.67
−6.134
1
29.86
H


ATOM
2201
OG1
THR
H
70
−17.202
9.186
−5.199
1
33.81
H


ATOM
2202
CG2
THR
H
70
−18.223
11.177
−6.085
1
30.79
H


ATOM
2203
C
THR
H
70
−19.659
7.668
−6.345
1
27.47
H


ATOM
2204
O
THR
H
70
−19.368
6.715
−5.632
1
27.45
H


ATOM
2205
N
ALA
H
71
−20.09
7.529
−7.589
1
30.73
H


ATOM
2206
CA
ALA
H
71
−20.198
6.211
−8.195
1
32.03
H


ATOM
2207
CB
ALA
H
71
−21.665
5.835
−8.408
1
32.14
H


ATOM
2208
C
ALA
H
71
−19.458
6.219
−9.52
1
32.58
H


ATOM
2209
O
ALA
H
71
−19.775
7
−10.425
1
32.18
H


ATOM
2210
N
ASP
H
72
−18.455
5.357
−9.616
1
33.49
H


ATOM
2211
CA
ASP
H
72
−17.664
5.24
−10.826
1
34.6
H


ATOM
2212
CB
ASP
H
72
−16.204
4.983
−10.472
1
34.52
H


ATOM
2213
CG
ASP
H
72
−15.298
5.046
−11.674
1
32.73
H


ATOM
2214
OD1
ASP
H
72
−15.522
4.283
−12.632
1
33.2
H


ATOM
2215
OD2
ASP
H
72
−14.357
5.863
−11.661
1
36.91
H


ATOM
2216
C
ASP
H
72
−18.232
4.065
−11.605
1
36.6
H


ATOM
2217
O
ASP
H
72
−17.828
2.916
−11.411
1
36.75
H


ATOM
2218
N
THR
H
73
−19.185
4.364
−12.478
1
37.82
H


ATOM
2219
CA
THR
H
73
−19.841
3.347
−13.284
1
39.48
H


ATOM
2220
CB
THR
H
73
−21.023
3.964
−14.058
1
40.05
H


ATOM
2221
OG1
THR
H
73
−20.617
5.211
−14.635
1
38.17
H


ATOM
2222
CG2
THR
H
73
−22.2
4.204
−13.126
1
38.01
H


ATOM
2223
C
THR
H
73
−18.881
2.658
−14.256
1
40.22
H


ATOM
2224
O
THR
H
73
−19.212
1.626
−14.848
1
42.22
H


ATOM
2225
N
SER
H
74
−17.693
3.229
−14.416
1
38.54
H


ATOM
2226
CA
SER
H
74
−16.698
2.651
−15.305
1
36.16
H


ATOM
2227
CB
SER
H
74
−15.635
3.679
−15.65
1
37.12
H


ATOM
2228
OG
SER
H
74
−16.167
4.648
−16.531
1
46.87
H


ATOM
2229
C
SER
H
74
−16.049
1.454
−14.645
1
35.12
H


ATOM
2230
O
SER
H
74
−15.804
0.435
−15.291
1
38.02
H


ATOM
2231
N
SER
H
75
−15.775
1.581
−13.351
1
30.84
H


ATOM
2232
CA
SER
H
75
−15.156
0.509
−12.591
1
26.56
H


ATOM
2233
CB
SER
H
75
−14.069
1.074
−11.686
1
24.36
H


ATOM
2234
OG
SER
H
75
−14.639
1.708
−10.557
1
23.71
H


ATOM
2235
C
SER
H
75
−16.197
−0.206
−11.729
1
26.8
H


ATOM
2236
O
SER
H
75
−15.883
−1.188
−11.061
1
24.01
H


ATOM
2237
N
ASN
H
76
−17.437
0.282
−11.758
1
27.22
H


ATOM
2238
CA
ASN
H
76
−18.507
−0.301
−10.949
1
27.68
H


ATOM
2239
CB
ASN
H
76
−18.804
−1.736
−11.393
1
27.7
H


ATOM
2240
CG
ASN
H
76
−19.486
−1.803
−12.751
1
31.28
H


ATOM
2241
OD1
ASN
H
76
−19.611
−2.877
−13.334
1
35.89
H


ATOM
2242
ND2
ASN
H
76
−19.94
−0.658
−13.255
1
30.38
H


ATOM
2243
C
ASN
H
76
−18.048
−0.276
−9.494
1
26.29
H


ATOM
2244
O
ASN
H
76
−17.971
−1.303
−8.818
1
24.22
H


ATOM
2245
N
THR
H
77
−17.728
0.921
−9.024
1
25.91
H


ATOM
2246
CA
THR
H
77
−17.263
1.097
−7.662
1
26.59
H


ATOM
2247
CB
THR
H
77
−15.713
1.275
−7.616
1
28
H


ATOM
2248
OG1
THR
H
77
−15.083
0.176
−8.291
1
27.85
H


ATOM
2249
CG2
THR
H
77
−15.213
1.321
−6.174
1
24.58
H


ATOM
2250
C
THR
H
77
−17.933
2.322
−7.066
1
24.64
H


ATOM
2251
O
THR
H
77
−18.065
3.357
−7.721
1
24.74
H


ATOM
2252
N
ALA
H
78
−18.383
2.182
−5.827
1
23.55
H


ATOM
2253
CA
ALA
H
78
−19.027
3.275
−5.122
1
21.51
H


ATOM
2254
CB
ALA
H
78
−20.329
2.797
−4.465
1
15.25
H


ATOM
2255
C
ALA
H
78
−18.029
3.719
−4.067
1
21.89
H


ATOM
2256
O
ALA
H
78
−17.408
2.895
−3.398
1
23.36
H


ATOM
2257
N
PHE
H
79
−17.858
5.02
−3.925
1
23.17
H


ATOM
2258
CA
PHE
H
79
−16.932
5.517
−2.935
1
24.04
H


ATOM
2259
CB
PHE
H
79
−15.829
6.338
−3.588
1
28.12
H


ATOM
2260
CG
PHE
H
79
−15.072
5.607
−4.654
1
28.58
H


ATOM
2261
CD1
PHE
H
79
−15.557
5.556
−5.957
1
28.92
H


ATOM
2262
CD2
PHE
H
79
−13.87
4.973
−4.356
1
28.25
H


ATOM
2263
CE1
PHE
H
79
−14.853
4.883
−6.955
1
32.13
H


ATOM
2264
CE2
PHE
H
79
−13.159
4.297
−5.343
1
31.64
H


ATOM
2265
CZ
PHE
H
79
−13.652
4.252
−6.648
1
29.74
H


ATOM
2266
C
PHE
H
79
−17.636
6.396
−1.939
1
25.52
H


ATOM
2267
O
PHE
H
79
−18.729
6.903
−2.189
1
27.79
H


ATOM
2268
N
MET
H
80
−17
6.553
−0.789
1
28.16
H


ATOM
2269
CA
MET
H
80
−17.506
7.425
0.249
1
27.74
H


ATOM
2270
CB
MET
H
80
−18.14
6.662
1.404
1
26.83
H


ATOM
2271
CG
MET
H
80
−18.645
7.625
2.487
1
26.82
H


ATOM
2272
SD
MET
H
80
−19.346
6.844
3.937
1
28.75
H


ATOM
2273
CE
MET
H
80
−17.861
5.956
4.567
1
25.98
H


ATOM
2274
C
MET
H
80
−16.321
8.209
0.772
1
28.71
H


ATOM
2275
O
MET
H
80
−15.344
7.634
1.246
1
27.48
H


ATOM
2276
N
GLN
H
81
−16.411
9.527
0.659
1
32.15
H


ATOM
2277
CA
GLN
H
81
−15.358
10.405
1.133
1
36.24
H


ATOM
2278
CB
GLN
H
81
−15.163
11.564
0.153
1
40.78
H


ATOM
2279
CG
GLN
H
81
−14.004
12.488
0.503
1
47.71
H


ATOM
2280
CD
GLN
H
81
−12.667
11.782
0.431
1
51.74
H


ATOM
2281
OE1
GLN
H
81
−12.281
11.268
−0.62
1
55.57
H


ATOM
2282
NE2
GLN
H
81
−11.951
11.747
1.551
1
54.75
H


ATOM
2283
C
GLN
H
81
−15.764
10.944
2.497
1
35.32
H


ATOM
2284
O
GLN
H
81
−16.841
11.519
2.639
1
35.15
H


ATOM
2285
N
LEU
H
82
−14.913
10.741
3.498
1
35.8
H


ATOM
2286
CA
LEU
H
82
−15.19
11.229
4.847
1
35.77
H


ATOM
2287
CB
LEU
H
82
−15.26
10.064
5.833
1
32.43
H


ATOM
2288
CG
LEU
H
82
−16.522
9.208
5.733
1
33.01
H


ATOM
2289
CD1
LEU
H
82
−16.386
7.982
6.624
1
31.81
H


ATOM
2290
CD2
LEU
H
82
−17.732
10.038
6.138
1
30.13
H


ATOM
2291
C
LEU
H
82
−14.116
12.215
5.289
1
37.04
H


ATOM
2292
O
LEU
H
82
−12.984
11.823
5.571
1
36.26
H


ATOM
2293
N
SER
H
82A
−14.474
13.494
5.35
1
38.53
H


ATOM
2294
CA
SER
H
82A
−13.529
14.526
5.753
1
41.94
H


ATOM
2295
CB
SER
H
82A
−13.593
15.703
4.777
1
43.28
H


ATOM
2296
OG
SER
H
82A
−14.832
16.381
4.881
1
50.55
H


ATOM
2297
C
SER
H
82A
−13.764
15.018
7.185
1
42.59
H


ATOM
2298
O
SER
H
82A
−14.8
14.733
7.793
1
37.55
H


ATOM
2299
N
SER
H
82B
−12.784
15.761
7.704
1
45.27
H


ATOM
2300
CA
SER
H
82B
−12.815
16.305
9.062
1
47.02
H


ATOM
2301
CB
SER
H
82B
−13.83
17.45
9.154
1
49.74
H


ATOM
2302
OG
SER
H
82B
−15.162
16.976
9.048
1
55.67
H


ATOM
2303
C
SER
H
82B
−13.158
15.21
10.077
1
45.72
H


ATOM
2304
O
SER
H
82B
−14.092
15.342
10.87
1
44.1
H


ATOM
2305
N
LEU
H
82C
−12.38
14.132
10.044
1
45.48
H


ATOM
2306
CA
LEU
H
82C
−12.582
12.989
10.93
1
45.09
H


ATOM
2307
CB
LEU
H
82C
−11.623
11.86
10.548
1
45.44
H


ATOM
2308
CG
LEU
H
82C
−12.191
10.823
9.573
1
44.73
H


ATOM
2309
CD1
LEU
H
82C
−13.207
11.462
8.643
1
43.75
H


ATOM
2310
CD2
LEU
H
82C
−11.044
10.196
8.79
1
46.77
H


ATOM
2311
C
LEU
H
82C
−12.456
13.283
12.413
1
43.98
H


ATOM
2312
O
LEU
H
82C
−11.649
14.108
12.829
1
44.39
H


ATOM
2313
N
THR
H
83
−13.26
12.569
13.195
1
43.97
H


ATOM
2314
CA
THR
H
83
−13.313
12.707
14.643
1
44.91
H


ATOM
2315
CB
THR
H
83
−14.572
13.475
15.042
1
46.59
H


ATOM
2316
OG1
THR
H
83
−14.476
14.816
14.548
1
46.18
H


ATOM
2317
CG2
THR
H
83
−14.751
13.485
16.558
1
49.35
H


ATOM
2318
C
THR
H
83
−13.348
11.337
15.315
1
46.38
H


ATOM
2319
O
THR
H
83
−13.652
10.333
14.674
1
48.88
H


ATOM
2320
N
SER
H
84
−13.042
11.292
16.607
1
46.34
H


ATOM
2321
CA
SER
H
84
−13.065
10.029
17.323
1
45.35
H


ATOM
2322
CB
SER
H
84
−12.639
10.231
18.779
1
45.13
H


ATOM
2323
OG
SER
H
84
−13.582
11.007
19.494
1
45.06
H


ATOM
2324
C
SER
H
84
−14.493
9.5
17.258
1
46.01
H


ATOM
2325
O
SER
H
84
−14.739
8.311
17.463
1
46.32
H


ATOM
2326
N
GLU
H
85
−15.428
10.398
16.959
1
45.5
H


ATOM
2327
CA
GLU
H
85
−16.842
10.051
16.858
1
45.98
H


ATOM
2328
CB
GLU
H
85
−17.688
11.331
16.825
1
50.25
H


ATOM
2329
CG
GLU
H
85
−19.179
11.117
16.555
1
54.41
H


ATOM
2330
CD
GLU
H
85
−20.058
11.461
17.751
1
58.62
H


ATOM
2331
OE1
GLU
H
85
−19.977
10.756
18.784
1
60.55
H


ATOM
2332
OE2
GLU
H
85
−20.831
12.44
17.657
1
59.71
H


ATOM
2333
C
GLU
H
85
−17.117
9.218
15.608
1
43.77
H


ATOM
2334
O
GLU
H
85
−18.03
8.391
15.589
1
44.09
H


ATOM
2335
N
ASP
H
86
−16.322
9.441
14.567
1
41.14
H


ATOM
2336
CA
ASP
H
86
−16.479
8.721
13.304
1
37.01
H


ATOM
2337
CB
ASP
H
86
−15.949
9.564
12.14
1
39.51
H


ATOM
2338
CG
ASP
H
86
−16.466
10.986
12.167
1
42.15
H


ATOM
2339
OD1
ASP
H
86
−17.64
11.187
12.54
1
43.42
H


ATOM
2340
OD2
ASP
H
86
−15.701
11.904
11.807
1
42.9
H


ATOM
2341
C
ASP
H
86
−15.769
7.369
13.299
1
33.42
H


ATOM
2342
O
ASP
H
86
−15.824
6.639
12.313
1
30.78
H


ATOM
2343
N
SER
H
87
−15.086
7.035
14.387
1
29.56
H


ATOM
2344
CA
SER
H
87
−14.409
5.748
14.441
1
28.75
H


ATOM
2345
CB
SER
H
87
−13.499
5.653
15.667
1
27.53
H


ATOM
2346
OG
SER
H
87
−12.347
6.458
15.496
1
31.1
H


ATOM
2347
C
SER
H
87
−15.469
4.668
14.499
1
25.65
H


ATOM
2348
O
SER
H
87
−16.296
4.655
15.412
1
23.46
H


ATOM
2349
N
ALA
H
88
−15.451
3.776
13.514
1
24
H


ATOM
2350
CA
ALA
H
88
−16.426
2.692
13.467
1
24.79
H


ATOM
2351
CB
ALA
H
88
−17.845
3.268
13.44
1
24.51
H


ATOM
2352
C
ALA
H
88
−16.239
1.756
12.279
1
23.07
H


ATOM
2353
O
ALA
H
88
−15.384
1.966
11.417
1
21.27
H


ATOM
2354
N
VAL
H
89
−17.042
0.701
12.255
1
22.59
H


ATOM
2355
CA
VAL
H
89
−16.996
−0.229
11.145
1
24.19
H


ATOM
2356
CB
VAL
H
89
−17.445
−1.654
11.572
1
24.22
H


ATOM
2357
CG1
VAL
H
89
−18.787
−1.585
12.283
1
31.12
H


ATOM
2358
CG2
VAL
H
89
−17.549
−2.559
10.354
1
20.23
H


ATOM
2359
C
VAL
H
89
−17.968
0.359
10.12
1
23.16
H


ATOM
2360
O
VAL
H
89
−19.111
0.696
10.443
1
18.77
H


ATOM
2361
N
TYR
H
90
−17.498
0.539
8.896
1
22.89
H


ATOM
2362
CA
TYR
H
90
−18.368
1.086
7.875
1
23.13
H


ATOM
2363
CB
TYR
H
90
−17.703
2.253
7.164
1
22.49
H


ATOM
2364
CG
TYR
H
90
−17.6
3.455
8.044
1
23.74
H


ATOM
2365
CD1
TYR
H
90
−16.574
3.571
8.98
1
21.39
H


ATOM
2366
CE1
TYR
H
90
−16.523
4.656
9.845
1
24.6
H


ATOM
2367
CD2
TYR
H
90
−18.572
4.452
7.99
1
22.12
H


ATOM
2368
CE2
TYR
H
90
−18.533
5.531
8.846
1
23.11
H


ATOM
2369
CZ
TYR
H
90
−17.511
5.632
9.771
1
23.88
H


ATOM
2370
OH
TYR
H
90
−17.49
6.709
10.623
1
26.15
H


ATOM
2371
C
TYR
H
90
−18.734
0.023
6.878
1
23.35
H


ATOM
2372
O
TYR
H
90
−17.863
−0.643
6.318
1
23.56
H


ATOM
2373
N
TYR
H
91
−20.031
−0.139
6.667
1
23.24
H


ATOM
2374
CA
TYR
H
91
−20.512
−1.133
5.731
1
25.43
H


ATOM
2375
CB
TYR
H
91
−21.67
−1.931
6.329
1
24.11
H


ATOM
2376
CG
TYR
H
91
−21.256
−2.896
7.407
1
22.57
H


ATOM
2377
CD1
TYR
H
91
−20.497
−4.017
7.103
1
21.38
H


ATOM
2378
CE1
TYR
H
91
−20.132
−4.916
8.089
1
25.79
H


ATOM
2379
CD2
TYR
H
91
−21.638
−2.694
8.729
1
21.45
H


ATOM
2380
CE2
TYR
H
91
−21.278
−3.585
9.725
1
23.8
H


ATOM
2381
CZ
TYR
H
91
−20.527
−4.695
9.401
1
25.11
H


ATOM
2382
OH
TYR
H
91
−20.162
−5.583
10.389
1
30.55
H


ATOM
2383
C
TYR
H
91
−20.972
−0.544
4.422
1
26.48
H


ATOM
2384
O
TYR
H
91
−21.47
0.582
4.355
1
27.26
H


ATOM
2385
N
CYS
H
92
−20.789
−1.338
3.381
1
27.58
H


ATOM
2386
CA
CYS
H
92
−21.213
−0.985
2.051
1
27.08
H


ATOM
2387
C
CYS
H
92
−22.366
−1.949
1.819
1
24.59
H


ATOM
2388
O
CYS
H
92
−22.194
−3.161
1.958
1
25.45
H


ATOM
2389
CB
CYS
H
92
−20.099
−1.255
1.039
1
29.34
H


ATOM
2390
SG
CYS
H
92
−20.701
−0.853
−0.617
1
40.33
H


ATOM
2391
N
ALA
H
93
−23.54
−1.435
1.485
1
20.72
H


ATOM
2392
CA
ALA
H
93
−24.677
−2.325
1.259
1
20.15
H


ATOM
2393
CB
ALA
H
93
−25.653
−2.239
2.431
1
21.64
H


ATOM
2394
C
ALA
H
93
−25.4
−2.01
−0.03
1
16.99
H


ATOM
2395
O
ALA
H
93
−25.338
−0.891
−0.525
1
19.38
H


ATOM
2396
N
ARG
H
94
−26.09
−2.995
−0.582
1
14.06
H


ATOM
2397
CA
ARG
H
94
−26.826
−2.755
−1.809
1
14.37
H


ATOM
2398
CB
ARG
H
94
−26.552
−3.861
−2.813
1
15.99
H


ATOM
2399
CG
ARG
H
94
−27.17
−3.601
−4.169
1
14.05
H


ATOM
2400
CD
ARG
H
94
−28.155
−4.682
−4.468
1
17.49
H


ATOM
2401
NE
ARG
H
94
−27.842
−5.38
−5.699
1
19.37
H


ATOM
2402
CZ
ARG
H
94
−28.383
−6.545
−6.036
1
21.79
H


ATOM
2403
NH1
ARG
H
94
−29.248
−7.129
−5.216
1
16.63
H


ATOM
2404
NH2
ARG
H
94
−28.088
−7.107
−7.205
1
24.27
H


ATOM
2405
C
ARG
H
94
−28.312
−2.687
−1.511
1
14.69
H


ATOM
2406
O
ARG
H
94
−28.87
−3.613
−0.927
1
13.41
H


ATOM
2407
N
SER
H
95
−28.946
−1.586
−1.906
1
15.09
H


ATOM
2408
CA
SER
H
95
−30.377
−1.404
−1.676
1
16.18
H


ATOM
2409
CB
SER
H
95
−30.761
0.066
−1.812
1
16.17
H


ATOM
2410
OG
SER
H
95
−32.091
0.272
−1.38
1
17.85
H


ATOM
2411
C
SER
H
95
−31.17
−2.235
−2.674
1
17.05
H


ATOM
2412
O
SER
H
95
−30.996
−2.101
−3.884
1
18.33
H


ATOM
2413
N
ALA
H
96
−32.047
−3.085
−2.15
1
16.69
H


ATOM
2414
CA
ALA
H
96
−32.86
−3.979
−2.962
1
16.13
H


ATOM
2415
CB
ALA
H
96
−33.622
−4.935
−2.053
1
15.97
H


ATOM
2416
C
ALA
H
96
−33.829
−3.303
−3.922
1
15.41
H


ATOM
2417
O
ALA
H
96
−34.273
−2.177
−3.707
1
16.59
H


ATOM
2418
N
SER
H
97
−34.164
−4.013
−4.988
1
15.94
H


ATOM
2419
CA
SER
H
97
−35.086
−3.48
−5.972
1
21.08
H


ATOM
2420
CB
SER
H
97
−35.283
−4.47
−7.123
1
19.36
H


ATOM
2421
OG
SER
H
97
−34.133
−4.546
−7.936
1
19.33
H


ATOM
2422
C
SER
H
97
−36.438
−3.18
−5.347
1
22.46
H


ATOM
2423
O
SER
H
97
−37.103
−2.233
−5.748
1
23.48
H


ATOM
2424
N
TYR
H
98
−36.828
−3.987
−4.361
1
23.91
H


ATOM
2425
CA
TYR
H
98
−38.121
−3.839
−3.706
1
24.89
H


ATOM
2426
CB
TYR
H
98
−38.683
−5.215
−3.359
1
25.72
H


ATOM
2427
CG
TYR
H
98
−39.358
−5.903
−4.519
1
29.65
H


ATOM
2428
CD1
TYR
H
98
−40.44
−5.312
−5.171
1
31.3
H


ATOM
2429
CE1
TYR
H
98
−41.077
−5.95
−6.237
1
31.57
H


ATOM
2430
CD2
TYR
H
98
−38.927
−7.15
−4.962
1
29.26
H


ATOM
2431
CE2
TYR
H
98
−39.555
−7.795
−6.025
1
29.82
H


ATOM
2432
CZ
TYR
H
98
−40.627
−7.192
−6.658
1
31.67
H


ATOM
2433
OH
TYR
H
98
−41.246
−7.837
−7.711
1
32.36
H


ATOM
2434
C
TYR
H
98
−38.203
−2.952
−2.474
1
25.8
H


ATOM
2435
O
TYR
H
98
−39.141
−3.081
−1.684
1
22.28
H


ATOM
2436
N
GLY
H
99
−37.243
−2.049
−2.301
1
26.59
H


ATOM
2437
CA
GLY
H
99
−37.304
−1.168
−1.149
1
26.74
H


ATOM
2438
C
GLY
H
99
−35.972
−0.741
−0.574
1
28.24
H


ATOM
2439
O
GLY
H
99
−34.922
−1.275
−0.933
1
27.76
H


ATOM
2440
N
ASP
H
100
−36.021
0.22
0.342
1
27.68
H


ATOM
2441
CA
ASP
H
100
−34.808
0.724
0.959
1
27.85
H


ATOM
2442
CB
ASP
H
100
−35.036
2.139
1.488
1
27.83
H


ATOM
2443
CG
ASP
H
100
−35.313
3.13
0.376
1
28.79
H


ATOM
2444
OD1
ASP
H
100
−36.389
3.037
−0.246
1
33.65
H


ATOM
2445
OD2
ASP
H
100
−34.457
3.997
0.114
1
28.79
H


ATOM
2446
C
ASP
H
100
−34.216
−0.173
2.045
1
26.78
H


ATOM
2447
O
ASP
H
100
−33.776
0.289
3.096
1
27.25
H


ATOM
2448
N
TYR
H
100A
−34.22
−1.471
1.785
1
25.66
H


ATOM
2449
CA
TYR
H
100A
−33.607
−2.421
2.7
1
22.34
H


ATOM
2450
CB
TYR
H
100A
−34.597
−3.507
3.134
1
19.31
H


ATOM
2451
CG
TYR
H
100A
−35.24
−4.243
1.997
1
22.22
H


ATOM
2452
CD1
TYR
H
100A
−36.344
−3.713
1.342
1
22.59
H


ATOM
2453
CE1
TYR
H
100A
−36.938
−4.379
0.29
1
22.5
H


ATOM
2454
CD2
TYR
H
100A
−34.74
−5.47
1.564
1
23.36
H


ATOM
2455
CE2
TYR
H
100A
−35.329
−6.147
0.509
1
23.22
H


ATOM
2456
CZ
TYR
H
100A
−36.432
−5.592
−0.124
1
24.18
H


ATOM
2457
OH
TYR
H
100A
−37.034
−6.249
−1.175
1
24.68
H


ATOM
2458
C
TYR
H
100A
−32.449
−3.02
1.899
1
18.6
H


ATOM
2459
O
TYR
H
100A
−32.502
−3.078
0.664
1
13.67
H


ATOM
2460
N
ALA
H
100B
−31.394
−3.435
2.588
1
19.07
H


ATOM
2461
CA
ALA
H
100B
−30.236
−3.994
1.904
1
18.47
H


ATOM
2462
CB
ALA
H
100B
−28.956
−3.5
2.564
1
16.36
H


ATOM
2463
C
ALA
H
100B
−30.262
−5.512
1.875
1
17.86
H


ATOM
2464
O
ALA
H
100B
−30.508
−6.16
2.89
1
20.82
H


ATOM
2465
N
ASP
H
101
−30.014
−6.07
0.696
1
19.66
H


ATOM
2466
CA
ASP
H
101
−29.993
−7.515
0.516
1
23.51
H


ATOM
2467
CB
ASP
H
101
−30.811
−7.897
−0.715
1
23.84
H


ATOM
2468
CG
ASP
H
101
−30.437
−7.085
−1.935
1
25.73
H


ATOM
2469
OD1
ASP
H
101
−29.455
−6.311
−1.857
1
23.8
H


ATOM
2470
OD2
ASP
H
101
−31.126
−7.228
−2.971
1
23.43
H


ATOM
2471
C
ASP
H
101
−28.566
−8.072
0.392
1
24.71
H


ATOM
2472
O
ASP
H
101
−28.355
−9.288
0.423
1
26.02
H


ATOM
2473
N
TYR
H
102
−27.591
−7.181
0.253
1
24.58
H


ATOM
2474
CA
TYR
H
102
−26.193
−7.587
0.15
1
25.52
H


ATOM
2475
CB
TYR
H
102
−25.793
−7.757
−1.318
1
28.06
H


ATOM
2476
CG
TYR
H
102
−26.451
−8.936
−2.007
1
30.4
H


ATOM
2477
CD1
TYR
H
102
−25.934
−10.229
−1.877
1
28.54
H


ATOM
2478
CE1
TYR
H
102
−26.533
−11.317
−2.524
1
28.74
H


ATOM
2479
CD2
TYR
H
102
−27.592
−8.758
−2.799
1
30.81
H


ATOM
2480
CE2
TYR
H
102
−28.202
−9.84
−3.446
1
30.98
H


ATOM
2481
CZ
TYR
H
102
−27.666
−11.113
−3.306
1
30.96
H


ATOM
2482
OH
TYR
H
102
−28.267
−12.17
−3.956
1
29.64
H


ATOM
2483
C
TYR
H
102
−25.312
−6.54
0.818
1
23.98
H


ATOM
2484
O
TYR
H
102
−25.402
−5.348
0.521
1
25.69
H


ATOM
2485
N
TRP
H
103
−24.462
−7
1.728
1
23.67
H


ATOM
2486
CA
TRP
H
103
−23.566
−6.127
2.474
1
18.64
H


ATOM
2487
CB
TRP
H
103
−23.812
−6.265
3.974
1
18.93
H


ATOM
2488
CG
TRP
H
103
−25.21
−5.98
4.403
1
18.53
H


ATOM
2489
CD2
TRP
H
103
−25.632
−4.934
5.273
1
15.85
H


ATOM
2490
CE2
TRP
H
103
−27.032
−5.023
5.383
1
16.74
H


ATOM
2491
CE3
TRP
H
103
−24.96
−3.924
5.97
1
20.28
H


ATOM
2492
CD1
TRP
H
103
−26.342
−6.651
4.031
1
17.82
H


ATOM
2493
NE1
TRP
H
103
−27.44
−6.083
4.615
1
17.13
H


ATOM
2494
CZ2
TRP
H
103
−27.778
−4.138
6.16
1
16.77
H


ATOM
2495
CZ3
TRP
H
103
−25.701
−3.04
6.746
1
19.94
H


ATOM
2496
CH2
TRP
H
103
−27.095
−3.154
6.831
1
19.4
H


ATOM
2497
C
TRP
H
103
−22.129
−6.505
2.217
1
20.41
H


ATOM
2498
O
TRP
H
103
−21.822
−7.672
1.95
1
20.83
H


ATOM
2499
N
GLY
H
104
−21.246
−5.514
2.287
1
21.1
H


ATOM
2500
CA
GLY
H
104
−19.833
−5.786
2.12
1
23.14
H


ATOM
2501
C
GLY
H
104
−19.396
−6.261
3.494
1
24.35
H


ATOM
2502
O
GLY
H
104
−20.184
−6.201
4.444
1
22.07
H


ATOM
2503
N
HIS
H
105
−18.162
−6.723
3.63
1
27.31
H


ATOM
2504
CA
HIS
H
105
−17.715
−7.197
4.933
1
31.34
H


ATOM
2505
CB
HIS
H
105
−16.574
−8.196
4.761
1
36.95
H


ATOM
2506
CG
HIS
H
105
−16.958
−9.371
3.922
1
45.33
H


ATOM
2507
CD2
HIS
H
105
−16.395
−9.899
2.81
1
47.96
H


ATOM
2508
ND1
HIS
H
105
−18.115
−10.087
4.146
1
47.69
H


ATOM
2509
CE1
HIS
H
105
−18.251
−11.003
3.203
1
51.12
H


ATOM
2510
NE2
HIS
H
105
−17.221
−10.91
2.379
1
50.5
H


ATOM
2511
C
HIS
H
105
−17.322
−6.069
5.865
1
28.34
H


ATOM
2512
O
HIS
H
105
−16.789
−6.299
6.942
1
29.33
H


ATOM
2513
N
GLY
H
106
−17.607
−4.846
5.447
1
27.15
H


ATOM
2514
CA
GLY
H
106
−17.289
−3.704
6.273
1
28.86
H


ATOM
2515
C
GLY
H
106
−15.811
−3.387
6.36
1
28.45
H


ATOM
2516
O
GLY
H
106
−14.959
−4.274
6.39
1
30.58
H


ATOM
2517
N
THR
H
107
−15.508
−2.098
6.376
1
28.87
H


ATOM
2518
CA
THR
H
107
−14.137
−1.637
6.497
1
28.05
H


ATOM
2519
CB
THR
H
107
−13.746
−0.72
5.32
1
26.06
H


ATOM
2520
OG1
THR
H
107
−12.69
0.147
5.731
1
31.24
H


ATOM
2521
CG2
THR
H
107
−14.918
0.102
4.862
1
28.02
H


ATOM
2522
C
THR
H
107
−14.072
−0.878
7.82
1
26.61
H


ATOM
2523
O
THR
H
107
−14.914
−0.022
8.09
1
25.66
H


ATOM
2524
N
THR
H
108
−13.1
−1.223
8.661
1
25.86
H


ATOM
2525
CA
THR
H
108
−12.962
−0.572
9.961
1
23.72
H


ATOM
2526
CB
THR
H
108
−12.349
−1.526
11.023
1
23.76
H


ATOM
2527
OG1
THR
H
108
−13.217
−2.653
11.232
1
22.42
H


ATOM
2528
CG2
THR
H
108
−12.164
−0.797
12.342
1
19.8
H


ATOM
2529
C
THR
H
108
−12.111
0.693
9.888
1
24.39
H


ATOM
2530
O
THR
H
108
−10.968
0.679
9.428
1
24.52
H


ATOM
2531
N
LEU
H
109
−12.691
1.793
10.345
1
23.92
H


ATOM
2532
CA
LEU
H
109
−12.013
3.071
10.352
1
23.67
H


ATOM
2533
CB
LEU
H
109
−12.832
4.098
9.575
1
23.95
H


ATOM
2534
CG
LEU
H
109
−12.365
5.549
9.684
1
23.82
H


ATOM
2535
CD1
LEU
H
109
−10.918
5.672
9.22
1
25.23
H


ATOM
2536
CD2
LEU
H
109
−13.274
6.423
8.847
1
22.33
H


ATOM
2537
C
LEU
H
109
−11.814
3.544
11.789
1
25.63
H


ATOM
2538
O
LEU
H
109
−12.765
3.647
12.568
1
25.05
H


ATOM
2539
N
THR
H
110
−10.565
3.826
12.135
1
26.28
H


ATOM
2540
CA
THR
H
110
−10.237
4.287
13.472
1
27.28
H


ATOM
2541
CB
THR
H
110
−9.26
3.317
14.176
1
29.45
H


ATOM
2542
OG1
THR
H
110
−9.753
1.97
14.081
1
30.69
H


ATOM
2543
CG2
THR
H
110
−9.11
3.706
15.648
1
27.31
H


ATOM
2544
C
THR
H
110
−9.596
5.669
13.375
1
26.95
H


ATOM
2545
O
THR
H
110
−8.536
5.838
12.778
1
25.4
H


ATOM
2546
N
VAL
H
111
−10.263
6.66
13.953
1
27.85
H


ATOM
2547
CA
VAL
H
111
−9.764
8.022
13.937
1
26.78
H


ATOM
2548
CB
VAL
H
111
−10.923
9.033
13.872
1
28.9
H


ATOM
2549
CG1
VAL
H
111
−10.371
10.45
13.829
1
24.78
H


ATOM
2550
CG2
VAL
H
111
−11.798
8.749
12.646
1
23.25
H


ATOM
2551
C
VAL
H
111
−8.991
8.215
15.227
1
30.11
H


ATOM
2552
O
VAL
H
111
−9.579
8.284
16.31
1
31.03
H


ATOM
2553
N
SER
H
112
−7.67
8.296
15.107
1
31.29
H


ATOM
2554
CA
SER
H
112
−6.802
8.458
16.265
1
32.03
H


ATOM
2555
CB
SER
H
112
−6.668
7.117
16.999
1
33.27
H


ATOM
2556
OG
SER
H
112
−5.6
7.124
17.933
1
35.37
H


ATOM
2557
C
SER
H
112
−5.425
8.947
15.848
1
32.3
H


ATOM
2558
O
SER
H
112
−4.954
8.643
14.748
1
31.76
H


ATOM
2559
N
SER
H
113
−4.787
9.703
16.736
1
31.76
H


ATOM
2560
CA
SER
H
113
−3.45
10.23
16.489
1
31.47
H


ATOM
2561
CB
SER
H
113
−3.233
11.512
17.308
1
33.88
H


ATOM
2562
OG
SER
H
113
−4.25
11.685
18.29
1
38.4
H


ATOM
2563
C
SER
H
113
−2.39
9.179
16.843
1
30.34
H


ATOM
2564
O
SER
H
113
−1.197
9.4
16.676
1
28.95
H


ATOM
2565
N
ALA
H
114
−2.843
8.023
17.316
1
29.4
H


ATOM
2566
CA
ALA
H
114
−1.938
6.945
17.684
1
28.43
H


ATOM
2567
CB
ALA
H
114
−2.665
5.93
18.565
1
25.6
H


ATOM
2568
C
ALA
H
114
−1.399
6.262
16.439
1
28.8
H


ATOM
2569
O
ALA
H
114
−2.036
6.276
15.392
1
32.36
H


ATOM
2570
N
LYS
H
115
−0.226
5.656
16.561
1
30.91
H


ATOM
2571
CA
LYS
H
115
0.387
4.952
15.445
1
32.2
H


ATOM
2572
CB
LYS
H
115
1.913
5.098
15.517
1
36.6
H


ATOM
2573
CG
LYS
H
115
2.399
6.456
15.013
1
41.36
H


ATOM
2574
CD
LYS
H
115
3.852
6.753
15.357
1
43.53
H


ATOM
2575
CE
LYS
H
115
4.287
8.076
14.725
1
45.01
H


ATOM
2576
NZ
LYS
H
115
3.399
9.23
15.08
1
47.73
H


ATOM
2577
C
LYS
H
115
−0.011
3.472
15.336
1
31.8
H


ATOM
2578
O
LYS
H
115
−0.237
2.776
16.334
1
29.54
H


ATOM
2579
N
THR
H
116
−0.1
3.009
14.094
1
32.32
H


ATOM
2580
CA
THR
H
116
−0.472
1.636
13.791
1
31.75
H


ATOM
2581
CB
THR
H
116
−0.538
1.424
12.278
1
30.67
H


ATOM
2582
OG1
THR
H
116
−1.555
2.269
11.732
1
33.19
H


ATOM
2583
CG2
THR
H
116
−0.836
−0.035
11.951
1
29.22
H


ATOM
2584
C
THR
H
116
0.525
0.644
14.366
1
32.75
H


ATOM
2585
O
THR
H
116
1.715
0.705
14.064
1
32.95
H


ATOM
2586
N
THR
H
117
0.035
−0.273
15.188
1
32.66
H


ATOM
2587
CA
THR
H
117
0.903
−1.276
15.783
1
32.97
H


ATOM
2588
CB
THR
H
117
0.99
−1.149
17.328
1
34.5
H


ATOM
2589
OG1
THR
H
117
1.269
0.205
17.7
1
37.69
H


ATOM
2590
CG2
THR
H
117
2.098
−2.04
17.86
1
35.92
H


ATOM
2591
C
THR
H
117
0.353
−2.657
15.495
1
30.42
H


ATOM
2592
O
THR
H
117
−0.766
−2.977
15.89
1
28.68
H


ATOM
2593
N
PRO
H
118
1.118
−3.488
14.777
1
29.41
H


ATOM
2594
CD
PRO
H
118
2.425
−3.275
14.131
1
28.81
H


ATOM
2595
CA
PRO
H
118
0.606
−4.832
14.507
1
27.06
H


ATOM
2596
CB
PRO
H
118
1.582
−5.365
13.462
1
28.68
H


ATOM
2597
CG
PRO
H
118
2.877
−4.689
13.835
1
27.03
H


ATOM
2598
C
PRO
H
118
0.694
−5.56
15.853
1
26.72
H


ATOM
2599
O
PRO
H
118
1.337
−5.067
16.784
1
27.22
H


ATOM
2600
N
PRO
H
119
0.049
−6.728
15.986
1
24.12
H


ATOM
2601
CD
PRO
H
119
−0.998
−7.346
15.148
1
21.57
H


ATOM
2602
CA
PRO
H
119
0.138
−7.404
17.278
1
22.05
H


ATOM
2603
CB
PRO
H
119
−1.245
−8.007
17.417
1
21.54
H


ATOM
2604
CG
PRO
H
119
−1.469
−8.515
16.01
1
20.32
H


ATOM
2605
C
PRO
H
119
1.209
−8.484
17.334
1
23.34
H


ATOM
2606
O
PRO
H
119
1.642
−9.011
16.31
1
23.27
H


ATOM
2607
N
SER
H
120
1.643
−8.81
18.541
1
23.35
H


ATOM
2608
CA
SER
H
120
2.604
−9.885
18.697
1
24.56
H


ATOM
2609
CB
SER
H
120
3.441
−9.714
19.965
1
20.54
H


ATOM
2610
OG
SER
H
120
4.305
−8.602
19.855
1
23.2
H


ATOM
2611
C
SER
H
120
1.694
−11.09
18.84
1
23.57
H


ATOM
2612
O
SER
H
120
0.739
−11.061
19.613
1
25.37
H


ATOM
2613
N
VAL
H
121
1.956
−12.131
18.071
1
22.62
H


ATOM
2614
CA
VAL
H
121
1.138
−13.314
18.165
1
22.29
H


ATOM
2615
CB
VAL
H
121
0.782
−13.868
16.779
1
23.47
H


ATOM
2616
CG1
VAL
H
121
−0.172
−15.053
16.922
1
19.89
H


ATOM
2617
CG2
VAL
H
121
0.153
−12.763
15.934
1
22.32
H


ATOM
2618
C
VAL
H
121
1.959
−14.319
18.929
1
23.79
H


ATOM
2619
O
VAL
H
121
2.981
−14.802
18.437
1
23
H


ATOM
2620
N
TYR
H
122
1.521
−14.609
20.148
1
24.01
H


ATOM
2621
CA
TYR
H
122
2.226
−15.552
20.99
1
25.51
H


ATOM
2622
CB
TYR
H
122
2.376
−14.988
22.404
1
24.61
H


ATOM
2623
CG
TYR
H
122
3.214
−13.731
22.451
1
30.94
H


ATOM
2624
CD1
TYR
H
122
4.501
−13.706
21.903
1
33.19
H


ATOM
2625
CE1
TYR
H
122
5.28
−12.544
21.93
1
30.97
H


ATOM
2626
CD2
TYR
H
122
2.727
−12.56
23.029
1
31.97
H


ATOM
2627
CE2
TYR
H
122
3.5
−11.393
23.06
1
31.89
H


ATOM
2628
CZ
TYR
H
122
4.774
−11.397
22.51
1
31.43
H


ATOM
2629
OH
TYR
H
122
5.545
−10.259
22.552
1
31.29
H


ATOM
2630
C
TYR
H
122
1.496
−16.877
21.025
1
24.8
H


ATOM
2631
O
TYR
H
122
0.276
−16.917
21.149
1
27.73
H


ATOM
2632
N
PRO
H
123
2.235
−17.985
20.879
1
23.29
H


ATOM
2633
CD
PRO
H
123
3.653
−18.086
20.478
1
20.29
H


ATOM
2634
CA
PRO
H
123
1.602
−19.301
20.909
1
21.41
H


ATOM
2635
CB
PRO
H
123
2.67
−20.204
20.297
1
22.22
H


ATOM
2636
CG
PRO
H
123
3.961
−19.541
20.731
1
17.63
H


ATOM
2637
C
PRO
H
123
1.236
−19.708
22.339
1
24.48
H


ATOM
2638
O
PRO
H
123
1.918
−19.34
23.302
1
25.06
H


ATOM
2639
N
LEU
H
124
0.148
−20.456
22.472
1
25.43
H


ATOM
2640
CA
LEU
H
124
−0.304
−20.923
23.769
1
26.22
H


ATOM
2641
CB
LEU
H
124
−1.696
−20.366
24.072
1
26.87
H


ATOM
2642
CG
LEU
H
124
−1.768
−19.04
24.844
1
28.04
H


ATOM
2643
CD1
LEU
H
124
−0.634
−18.111
24.435
1
25.09
H


ATOM
2644
CD2
LEU
H
124
−3.131
−18.392
24.589
1
27.98
H


ATOM
2645
C
LEU
H
124
−0.32
−22.442
23.785
1
28.92
H


ATOM
2646
O
LEU
H
124
−1.197
−23.083
23.186
1
28.5
H


ATOM
2647
N
ALA
H
125
0.67
−23.011
24.469
1
30.55
H


ATOM
2648
CA
ALA
H
125
0.806
−24.456
24.586
1
29.96
H


ATOM
2649
CB
ALA
H
125
1.972
−24.932
23.755
1
29.46
H


ATOM
2650
C
ALA
H
125
1.013
−24.848
26.038
1
32.47
H


ATOM
2651
O
ALA
H
125
1.637
−24.115
26.807
1
31.17
H


ATOM
2652
N
PRO
H
126
0.497
−26.021
26.428
1
35.22
H


ATOM
2653
CD
PRO
H
126
−0.212
−26.945
25.527
1
34.56
H


ATOM
2654
CA
PRO
H
126
0.584
−26.579
27.783
1
37.74
H


ATOM
2655
CB
PRO
H
126
−0.065
−27.952
27.63
1
36.57
H


ATOM
2656
CG
PRO
H
126
−1.025
−27.75
26.494
1
38.9
H


ATOM
2657
C
PRO
H
126
2.017
−26.683
28.324
1
41.77
H


ATOM
2658
O
PRO
H
126
2.971
−26.794
27.553
1
40.49
H


ATOM
2659
N
GLY
H
127
2.146
−26.662
29.652
1
46.28
H


ATOM
2660
CA
GLY
H
127
3.446
−26.755
30.3
1
50.84
H


ATOM
2661
C
GLY
H
127
4.123
−28.108
30.168
1
55.49
H


ATOM
2662
O
GLY
H
127
4.44
−28.526
29.061
1
56.73
H


ATOM
2663
N
CYS
H
128
4.349
−28.795
31.288
1
60.96
H


ATOM
2664
CA
CYS
H
128
5.01
−30.104
31.266
1
65.66
H


ATOM
2665
CB
CYS
H
128
6.336
−30.046
32.036
1
64.76
H


ATOM
2666
SG
CYS
H
128
7.689
−29.222
31.164
1
65.85
H


ATOM
2667
C
CYS
H
128
4.175
−31.268
31.806
1
68.44
H


ATOM
2668
O
CYS
H
128
3.593
−32.031
31.034
1
69.11
H


ATOM
2669
N
GLY
H
129
4.131
−31.406
33.13
1
71.63
H


ATOM
2670
CA
GLY
H
129
3.383
−32.491
33.75
1
76.46
H


ATOM
2671
C
GLY
H
129
1.88
−32.281
33.867
1
80.03
H


ATOM
2672
O
GLY
H
129
1.387
−31.785
34.885
1
79.58
H


ATOM
2673
N
ASP
H
130
1.149
−32.678
32.827
1
82.64
H


ATOM
2674
CA
ASP
H
130
−0.303
−32.529
32.803
1
84.12
H


ATOM
2675
CB
ASP
H
130
−0.686
−31.181
32.172
1
84.72
H


ATOM
2676
CG
ASP
H
130
0.001
−30.939
30.834
1
84.6
H


ATOM
2677
OD1
ASP
H
130
1.245
−30.81
30.815
1
82.66
H


ATOM
2678
OD2
ASP
H
130
−0.704
−30.877
29.802
1
85.79
H


ATOM
2679
C
ASP
H
130
−1.021
−33.657
32.062
1
84.81
H


ATOM
2680
O
ASP
H
130
−2.122
−34.055
32.453
1
85.5
H


ATOM
2681
N
THR
H
133
−0.394
−34.168
31.001
1
84.51
H


ATOM
2682
CA
THR
H
133
−0.971
−35.239
30.182
1
83.51
H


ATOM
2683
CB
THR
H
133
−0.541
−36.646
30.688
1
84.81
H


ATOM
2684
OG1
THR
H
133
−0.887
−36.796
32.072
1
87.07
H


ATOM
2685
CG2
THR
H
133
0.959
−36.833
30.52
1
84.91
H


ATOM
2686
C
THR
H
133
−2.498
−35.151
30.149
1
81.09
H


ATOM
2687
O
THR
H
133
−3.201
−36.091
30.529
1
82.19
H


ATOM
2688
N
THR
H
134
−2.99
−34.002
29.694
1
76.75
H


ATOM
2689
CA
THR
H
134
−4.418
−33.72
29.592
1
72.89
H


ATOM
2690
CB
THR
H
134
−4.664
−32.554
28.62
1
73.75
H


ATOM
2691
OG1
THR
H
134
−4.232
−32.931
27.307
1
74.08
H


ATOM
2692
CG2
THR
H
134
−3.879
−31.321
29.064
1
73.63
H


ATOM
2693
C
THR
H
134
−5.243
−34.924
29.137
1
69.33
H


ATOM
2694
O
THR
H
134
−4.703
−35.889
28.596
1
70.8
H


ATOM
2695
N
GLY
H
135
−6.556
−34.845
29.344
1
64.05
H


ATOM
2696
CA
GLY
H
135
−7.453
−35.933
28.982
1
55.58
H


ATOM
2697
C
GLY
H
135
−7.546
−36.338
27.521
1
50.06
H


ATOM
2698
O
GLY
H
135
−6.538
−36.564
26.854
1
47.47
H


ATOM
2699
N
SER
H
136
−8.774
−36.448
27.027
1
46.15
H


ATOM
2700
CA
SER
H
136
−9.015
−36.841
25.646
1
43.05
H


ATOM
2701
CB
SER
H
136
−10.43
−37.4
25.491
1
42.17
H


ATOM
2702
OG
SER
H
136
−10.499
−38.762
25.868
1
41.16
H


ATOM
2703
C
SER
H
136
−8.818
−35.697
24.659
1
41.99
H


ATOM
2704
O
SER
H
136
−8.761
−35.919
23.449
1
43.69
H


ATOM
2705
N
SER
H
137
−8.72
−34.474
25.164
1
37.32
H


ATOM
2706
CA
SER
H
137
−8.53
−33.34
24.279
1
34.59
H


ATOM
2707
CB
SER
H
137
−9.869
−32.663
23.999
1
33.93
H


ATOM
2708
OG
SER
H
137
−10.296
−31.9
25.112
1
35.89
H


ATOM
2709
C
SER
H
137
−7.562
−32.32
24.856
1
32.47
H


ATOM
2710
O
SER
H
137
−7.541
−32.082
26.063
1
33.35
H


ATOM
2711
N
VAL
H
138
−6.748
−31.725
23.991
1
29.19
H


ATOM
2712
CA
VAL
H
138
−5.811
−30.705
24.438
1
26.57
H


ATOM
2713
CB
VAL
H
138
−4.345
−31.029
24.023
1
25.65
H


ATOM
2714
CG1
VAL
H
138
−4.18
−30.917
22.523
1
26.61
H


ATOM
2715
CG2
VAL
H
138
−3.384
−30.086
24.728
1
25.78
H


ATOM
2716
C
VAL
H
138
−6.248
−29.374
23.825
1
23.93
H


ATOM
2717
O
VAL
H
138
−6.633
−29.31
22.662
1
22.96
H


ATOM
2718
N
THR
H
139
−6.222
−28.32
24.63
1
22.44
H


ATOM
2719
CA
THR
H
139
−6.605
−27
24.163
1
21.65
H


ATOM
2720
CB
THR
H
139
−7.536
−26.295
25.163
1
20.78
H


ATOM
2721
OG1
THR
H
139
−8.721
−27.081
25.352
1
21.51
H


ATOM
2722
CG2
THR
H
139
−7.911
−24.913
24.638
1
15.29
H


ATOM
2723
C
THR
H
139
−5.366
−26.139
23.978
1
21.78
H


ATOM
2724
O
THR
H
139
−4.546
−26.008
24.89
1
22.31
H


ATOM
2725
N
LEU
H
140
−5.231
−25.557
22.795
1
20.44
H


ATOM
2726
CA
LEU
H
140
−4.091
−24.703
22.501
1
21.47
H


ATOM
2727
CB
LEU
H
140
−3.319
−25.233
21.297
1
21.98
H


ATOM
2728
CG
LEU
H
140
−2.908
−26.703
21.275
1
22.97
H


ATOM
2729
CD1
LEU
H
140
−2.369
−27.067
19.885
1
19.48
H


ATOM
2730
CD2
LEU
H
140
−1.879
−26.955
22.353
1
21.81
H


ATOM
2731
C
LEU
H
140
−4.66
−23.344
22.157
1
21.57
H


ATOM
2732
O
LEU
H
140
−5.878
−23.143
22.219
1
18.79
H


ATOM
2733
N
GLY
H
141
−3.784
−22.42
21.774
1
20.53
H


ATOM
2734
CA
GLY
H
141
−4.253
−21.099
21.415
1
21.99
H


ATOM
2735
C
GLY
H
141
−3.185
−20.104
21.005
1
22.57
H


ATOM
2736
O
GLY
H
141
−2.008
−20.435
20.886
1
20.97
H


ATOM
2737
N
CYS
H
142
−3.624
−18.869
20.786
1
23.01
H


ATOM
2738
CA
CYS
H
142
−2.754
−17.775
20.399
1
22.78
H


ATOM
2739
C
CYS
H
142
−3.174
−16.516
21.129
1
23.48
H


ATOM
2740
O
CYS
H
142
−4.364
−16.221
21.259
1
24.59
H


ATOM
2741
CB
CYS
H
142
−2.826
−17.53
18.891
1
23.52
H


ATOM
2742
SG
CYS
H
142
−1.882
−18.718
17.881
1
28.33
H


ATOM
2743
N
LEU
H
143
−2.186
−15.786
21.624
1
24.17
H


ATOM
2744
CA
LEU
H
143
−2.432
−14.54
22.32
1
23.71
H


ATOM
2745
CB
LEU
H
143
−1.616
−14.482
23.607
1
21.28
H


ATOM
2746
CG
LEU
H
143
−1.7
−13.152
24.345
1
23.06
H


ATOM
2747
CD1
LEU
H
143
−3.089
−12.963
24.909
1
25.91
H


ATOM
2748
CD2
LEU
H
143
−0.677
−13.121
25.445
1
22.79
H


ATOM
2749
C
LEU
H
143
−1.984
−13.443
21.361
1
27.23
H


ATOM
2750
O
LEU
H
143
−0.808
−13.373
20.99
1
29.27
H


ATOM
2751
N
VAL
H
144
−2.936
−12.619
20.935
1
26.57
H


ATOM
2752
CA
VAL
H
144
−2.678
−11.515
20.015
1
24.48
H


ATOM
2753
CB
VAL
H
144
−3.864
−11.356
19.032
1
24.79
H


ATOM
2754
CG1
VAL
H
144
−3.539
−10.348
17.95
1
23.2
H


ATOM
2755
CG2
VAL
H
144
−4.189
−12.698
18.413
1
23.76
H


ATOM
2756
C
VAL
H
144
−2.553
−10.294
20.921
1
26.92
H


ATOM
2757
O
VAL
H
144
−3.554
−9.72
21.354
1
26.36
H


ATOM
2758
N
LYS
H
145
−1.316
−9.892
21.198
1
27.8
H


ATOM
2759
CA
LYS
H
145
−1.078
−8.792
22.12
1
29.63
H


ATOM
2760
CB
LYS
H
145
−0.23
−9.326
23.281
1
27.41
H


ATOM
2761
CG
LYS
H
145
0.097
−8.312
24.345
1
31.17
H


ATOM
2762
CD
LYS
H
145
0.872
−8.934
25.495
1
32.78
H


ATOM
2763
CE
LYS
H
145
1.2
−7.877
26.538
1
33.51
H


ATOM
2764
NZ
LYS
H
145
−0.031
−7.126
26.946
1
36.05
H


ATOM
2765
C
LYS
H
145
−0.475
−7.479
21.606
1
30.6
H


ATOM
2766
O
LYS
H
145
0.355
−7.464
20.7
1
31.17
H


ATOM
2767
N
GLY
H
146
−0.929
−6.384
22.215
1
30.75
H


ATOM
2768
CA
GLY
H
146
−0.443
−5.049
21.914
1
32.51
H


ATOM
2769
C
GLY
H
146
−0.56
−4.519
20.503
1
33.52
H


ATOM
2770
O
GLY
H
146
0.455
−4.241
19.865
1
34.32
H


ATOM
2771
N
TYR
H
147
−1.785
−4.341
20.02
1
31.35
H


ATOM
2772
CA
TYR
H
147
−1.978
−3.841
18.67
1
30.36
H


ATOM
2773
CB
TYR
H
147
−2.427
−4.984
17.753
1
28.17
H


ATOM
2774
CG
TYR
H
147
−3.806
−5.52
18.06
1
29.92
H


ATOM
2775
CD1
TYR
H
147
−4.94
−4.926
17.507
1
29.62
H


ATOM
2776
CE1
TYR
H
147
−6.212
−5.4
17.795
1
29.64
H


ATOM
2777
CD2
TYR
H
147
−3.984
−6.611
18.919
1
28.56
H


ATOM
2778
CE2
TYR
H
147
−5.255
−7.092
19.216
1
26.84
H


ATOM
2779
CZ
TYR
H
147
−6.362
−6.48
18.646
1
27.77
H


ATOM
2780
OH
TYR
H
147
−7.625
−6.943
18.909
1
29.25
H


ATOM
2781
C
TYR
H
147
−2.96
−2.672
18.596
1
30.43
H


ATOM
2782
O
TYR
H
147
−3.71
−2.405
19.543
1
29.48
H


ATOM
2783
N
PHE
H
148
−2.945
−1.977
17.461
1
29.2
H


ATOM
2784
CA
PHE
H
148
−3.818
−0.827
17.253
1
29.41
H


ATOM
2785
CB
PHE
H
148
−3.339
0.346
18.107
1
30.71
H


ATOM
2786
CG
PHE
H
148
−4.307
1.494
18.151
1
31.27
H


ATOM
2787
CD1
PHE
H
148
−5.406
1.459
19.005
1
30.32
H


ATOM
2788
CD2
PHE
H
148
−4.153
2.585
17.305
1
30.76
H


ATOM
2789
CE1
PHE
H
148
−6.328
2.489
19.019
1
28.57
H


ATOM
2790
CE2
PHE
H
148
−5.075
3.623
17.312
1
29.76
H


ATOM
2791
CZ
PHE
H
148
−6.166
3.573
18.169
1
28.87
H


ATOM
2792
C
PHE
H
148
−3.775
−0.428
15.787
1
29.92
H


ATOM
2793
O
PHE
H
148
−2.712
−0.497
15.162
1
30.58
H


ATOM
2794
N
PRO
H
149
−4.923
−0.034
15.211
1
30.8
H


ATOM
2795
CD
PRO
H
149
−4.966
0.398
13.804
1
31.07
H


ATOM
2796
CA
PRO
H
149
−6.25
0.059
15.824
1
31.64
H


ATOM
2797
CB
PRO
H
149
−7.014
0.904
14.828
1
30.75
H


ATOM
2798
CG
PRO
H
149
−6.451
0.416
13.522
1
31.45
H


ATOM
2799
C
PRO
H
149
−6.833
−1.338
15.96
1
35.7
H


ATOM
2800
O
PRO
H
149
−6.136
−2.324
15.721
1
37.22
H


ATOM
2801
N
GLU
H
150
−8.107
−1.447
16.319
1
38.82
H


ATOM
2802
CA
GLU
H
150
−8.674
−2.775
16.463
1
42.16
H


ATOM
2803
CB
GLU
H
150
−9.751
−2.82
17.547
1
43.28
H


ATOM
2804
CG
GLU
H
150
−9.651
−4.096
18.362
1
49.59
H


ATOM
2805
CD
GLU
H
150
−10.953
−4.471
18.99
1
52.69
H


ATOM
2806
OE1
GLU
H
150
−11.574
−3.578
19.603
1
58.52
H


ATOM
2807
OE2
GLU
H
150
−11.346
−5.656
18.883
1
53.82
H


ATOM
2808
C
GLU
H
150
−9.203
−3.446
15.2
1
42.64
H


ATOM
2809
O
GLU
H
150
−9.114
−2.903
14.088
1
38.81
H


ATOM
2810
N
SER
H
151
−9.756
−4.636
15.436
1
44.26
H


ATOM
2811
CA
SER
H
151
−10.273
−5.542
14.442
1
44.02
H


ATOM
2812
CB
SER
H
151
−10.998
−4.814
13.308
1
47.18
H


ATOM
2813
OG
SER
H
151
−11.139
−5.69
12.176
1
50.49
H


ATOM
2814
C
SER
H
151
−9.103
−6.356
13.874
1
42.18
H


ATOM
2815
O
SER
H
151
−8.257
−5.854
13.128
1
38.7
H


ATOM
2816
N
VAL
H
152
−9.038
−7.613
14.282
1
39.36
H


ATOM
2817
CA
VAL
H
152
−8.035
−8.526
13.775
1
36.34
H


ATOM
2818
CB
VAL
H
152
−6.922
−8.814
14.791
1
37.24
H


ATOM
2819
CG1
VAL
H
152
−6.175
−7.546
15.129
1
39.5
H


ATOM
2820
CG2
VAL
H
152
−7.497
−9.444
16.025
1
35.37
H


ATOM
2821
C
VAL
H
152
−8.816
−9.8
13.533
1
36.17
H


ATOM
2822
O
VAL
H
152
−9.867
−10.017
14.137
1
31.54
H


ATOM
2823
N
THR
H
153
−8.331
−10.634
12.631
1
35.21
H


ATOM
2824
CA
THR
H
153
−9.038
−11.865
12.361
1
36.21
H


ATOM
2825
CB
THR
H
153
−9.552
−11.904
10.919
1
38.68
H


ATOM
2826
OG1
THR
H
153
−10.67
−11.018
10.806
1
41.43
H


ATOM
2827
CG2
THR
H
153
−9.977
−13.317
10.527
1
39.83
H


ATOM
2828
C
THR
H
153
−8.118
−13.031
12.609
1
34.92
H


ATOM
2829
O
THR
H
153
−7.016
−13.088
12.071
1
33.33
H


ATOM
2830
N
VAL
H
154
−8.576
−13.947
13.451
1
31.5
H


ATOM
2831
CA
VAL
H
154
−7.809
−15.12
13.792
1
31.22
H


ATOM
2832
CB
VAL
H
154
−7.737
−15.309
15.303
1
30.78
H


ATOM
2833
CG1
VAL
H
154
−7.012
−16.601
15.62
1
27.95
H


ATOM
2834
CG2
VAL
H
154
−7.045
−14.121
15.938
1
29.29
H


ATOM
2835
C
VAL
H
154
−8.459
−16.346
13.193
1
32.63
H


ATOM
2836
O
VAL
H
154
−9.475
−16.839
13.69
1
33.63
H


ATOM
2837
N
THR
H
156
−7.872
−16.828
12.11
1
31.88
H


ATOM
2838
CA
THR
H
156
−8.369
−18.018
11.459
1
30.95
H


ATOM
2839
CB
THR
H
156
−8.482
−17.807
9.942
1
31.53
H


ATOM
2840
OG1
THR
H
156
−7.19
−17.529
9.396
1
36.55
H


ATOM
2841
CG2
THR
H
156
−9.389
−16.631
9.651
1
34.65
H


ATOM
2842
C
THR
H
156
−7.363
−19.12
11.786
1
29.23
H


ATOM
2843
O
THR
H
156
−6.154
−18.955
11.601
1
27.96
H


ATOM
2844
N
TRP
H
157
−7.872
−20.227
12.31
1
26.51
H


ATOM
2845
CA
TRP
H
157
−7.035
−21.349
12.68
1
24.63
H


ATOM
2846
CB
TRP
H
157
−7.557
−22.029
13.937
1
23.99
H


ATOM
2847
CG
TRP
H
157
−7.307
−21.306
15.21
1
22.27
H


ATOM
2848
CD2
TRP
H
157
−6.163
−21.45
16.058
1
24.4
H


ATOM
2849
CE2
TRP
H
157
−6.385
−20.644
17.196
1
23.46
H


ATOM
2850
CE3
TRP
H
157
−4.971
−22.187
15.967
1
22.9
H


ATOM
2851
CD1
TRP
H
157
−8.146
−20.44
15.84
1
21.83
H


ATOM
2852
NE1
TRP
H
157
−7.602
−20.04
17.037
1
25.72
H


ATOM
2853
CZ2
TRP
H
157
−5.462
−20.551
18.239
1
23.3
H


ATOM
2854
CZ3
TRP
H
157
−4.052
−22.094
17.004
1
24.96
H


ATOM
2855
CH2
TRP
H
157
−4.306
−21.279
18.128
1
24.38
H


ATOM
2856
C
TRP
H
157
−6.996
−22.387
11.583
1
27.71
H


ATOM
2857
O
TRP
H
157
−8.001
−22.652
10.919
1
26.57
H


ATOM
2858
N
ASN
H
162
−5.825
−22.984
11.405
1
30
H


ATOM
2859
CA
ASN
H
162
−5.647
−24.022
10.411
1
31.28
H


ATOM
2860
CB
ASN
H
162
−4.63
−23.565
9.367
1
38.76
H


ATOM
2861
CG
ASN
H
162
−4.665
−24.413
8.114
1
44.91
H


ATOM
2862
OD1
ASN
H
162
−5.676
−24.456
7.411
1
49.41
H


ATOM
2863
ND2
ASN
H
162
−3.56
−25.101
7.828
1
50.92
H


ATOM
2864
C
ASN
H
162
−5.144
−25.255
11.164
1
29.56
H


ATOM
2865
O
ASN
H
162
−4.057
−25.24
11.743
1
29.02
H


ATOM
2866
N
SER
H
163
−5.944
−26.315
11.182
1
28.36
H


ATOM
2867
CA
SER
H
163
−5.549
−27.535
11.884
1
27.36
H


ATOM
2868
CB
SER
H
163
−6.497
−27.805
13.059
1
28.61
H


ATOM
2869
OG
SER
H
163
−7.826
−28.039
12.615
1
30.36
H


ATOM
2870
C
SER
H
163
−5.524
−28.75
10.958
1
26.27
H


ATOM
2871
O
SER
H
163
−6.063
−28.714
9.851
1
23.64
H


ATOM
2872
N
GLY
H
164
−4.884
−29.82
11.422
1
26.21
H


ATOM
2873
CA
GLY
H
164
−4.797
−31.037
10.642
1
23.94
H


ATOM
2874
C
GLY
H
164
−5.895
−31.992
11.06
1
24.52
H


ATOM
2875
O
GLY
H
164
−6.263
−32.895
10.313
1
26.76
H


ATOM
2876
N
SER
H
165
−6.419
−31.787
12.262
1
23.72
H


ATOM
2877
CA
SER
H
165
−7.489
−32.624
12.796
1
24.33
H


ATOM
2878
CB
SER
H
165
−7.412
−32.669
14.325
1
22.62
H


ATOM
2879
OG
SER
H
165
−7.511
−31.362
14.873
1
24.65
H


ATOM
2880
C
SER
H
165
−8.84
−32.062
12.38
1
24.08
H


ATOM
2881
O
SER
H
165
−9.039
−30.845
12.388
1
25.83
H


ATOM
2882
N
LEU
H
166
−9.766
−32.944
12.017
1
24.1
H


ATOM
2883
CA
LEU
H
166
−11.101
−32.519
11.616
1
24.5
H


ATOM
2884
CB
LEU
H
166
−11.905
−33.705
11.085
1
24.2
H


ATOM
2885
CG
LEU
H
166
−11.383
−34.429
9.843
1
24.41
H


ATOM
2886
CD1
LEU
H
166
−12.384
−35.495
9.452
1
20.24
H


ATOM
2887
CD2
LEU
H
166
−11.178
−33.449
8.695
1
26.17
H


ATOM
2888
C
LEU
H
166
−11.821
−31.899
12.81
1
25.68
H


ATOM
2889
O
LEU
H
166
−12.782
−31.156
12.653
1
28.87
H


ATOM
2890
N
SER
H
167
−11.363
−32.223
14.011
1
27.47
H


ATOM
2891
CA
SER
H
167
−11.956
−31.656
15.214
1
30.09
H


ATOM
2892
CB
SER
H
167
−12.045
−32.696
16.338
1
28.81
H


ATOM
2893
OG
SER
H
167
−13.107
−33.609
16.121
1
28.47
H


ATOM
2894
C
SER
H
167
−11.088
−30.495
15.669
1
30.95
H


ATOM
2895
O
SER
H
167
−9.891
−30.66
15.913
1
30.82
H


ATOM
2896
N
SER
H
168
−11.698
−29.321
15.758
1
31.76
H


ATOM
2897
CA
SER
H
168
−11.011
−28.11
16.2
1
34.66
H


ATOM
2898
CB
SER
H
168
−10.246
−27.449
15.053
1
32.7
H


ATOM
2899
OG
SER
H
168
−9.8
−28.396
14.106
1
38.8
H


ATOM
2900
C
SER
H
168
−12.101
−27.156
16.639
1
36.62
H


ATOM
2901
O
SER
H
168
−12.705
−26.483
15.804
1
42.63
H


ATOM
2902
N
SER
H
169
−12.392
−27.108
17.928
1
35.82
H


ATOM
2903
CA
SER
H
169
−13.41
−26.18
18.386
1
33.04
H


ATOM
2904
CB
SER
H
169
−14.198
−26.779
19.558
1
35.47
H


ATOM
2905
OG
SER
H
169
−13.365
−27.141
20.646
1
40.34
H


ATOM
2906
C
SER
H
169
−12.674
−24.903
18.79
1
29.06
H


ATOM
2907
O
SER
H
169
−11.85
−24.913
19.703
1
28.2
H


ATOM
2908
N
VAL
H
171
−12.942
−23.812
18.081
1
25.62
H


ATOM
2909
CA
VAL
H
171
−12.27
−22.564
18.391
1
26.6
H


ATOM
2910
CB
VAL
H
171
−11.68
−21.886
17.117
1
25.15
H


ATOM
2911
CG1
VAL
H
171
−11.072
−22.931
16.189
1
19.83
H


ATOM
2912
CG2
VAL
H
171
−12.742
−21.082
16.418
1
25.61
H


ATOM
2913
C
VAL
H
171
−13.162
−21.552
19.103
1
26.99
H


ATOM
2914
O
VAL
H
171
−14.359
−21.458
18.848
1
26.41
H


ATOM
2915
N
HIS
H
172
−12.552
−20.809
20.018
1
28.53
H


ATOM
2916
CA
HIS
H
172
−13.241
−19.774
20.766
1
28.37
H


ATOM
2917
CB
HIS
H
172
−13.294
−20.102
22.263
1
27.07
H


ATOM
2918
CG
HIS
H
172
−14.108
−21.313
22.588
1
26.96
H


ATOM
2919
CD2
HIS
H
172
−13.774
−22.471
23.204
1
27.44
H


ATOM
2920
ND1
HIS
H
172
−15.438
−21.425
22.252
1
27.1
H


ATOM
2921
CE1
HIS
H
172
−15.89
−22.603
22.646
1
27.32
H


ATOM
2922
NE2
HIS
H
172
−14.901
−23.256
23.225
1
28.19
H


ATOM
2923
C
HIS
H
172
−12.418
−18.523
20.57
1
28.96
H


ATOM
2924
O
HIS
H
172
−11.214
−18.513
20.828
1
30.62
H


ATOM
2925
N
THR
H
173
−13.057
−17.474
20.084
1
28.64
H


ATOM
2926
CA
THR
H
173
−12.364
−16.222
19.891
1
29.04
H


ATOM
2927
CB
THR
H
173
−12.64
−15.666
18.498
1
29.22
H


ATOM
2928
OG1
THR
H
173
−12.194
−16.619
17.527
1
29.59
H


ATOM
2929
CG2
THR
H
173
−11.893
−14.358
18.289
1
29.24
H


ATOM
2930
C
THR
H
173
−12.858
−15.263
20.969
1
28.6
H


ATOM
2931
O
THR
H
173
−14.058
−15.037
21.114
1
28.8
H


ATOM
2932
N
PHE
H
174
−11.925
−14.717
21.736
1
27.18
H


ATOM
2933
CA
PHE
H
174
−12.274
−13.813
22.814
1
29.8
H


ATOM
2934
CB
PHE
H
174
−11.43
−14.145
24.043
1
31.04
H


ATOM
2935
CG
PHE
H
174
−11.466
−15.599
24.417
1
33.26
H


ATOM
2936
CD1
PHE
H
174
−10.714
−16.533
23.707
1
34.51
H


ATOM
2937
CD2
PHE
H
174
−12.283
−16.044
25.451
1
33.41
H


ATOM
2938
CE1
PHE
H
174
−10.774
−17.889
24.018
1
33.07
H


ATOM
2939
CE2
PHE
H
174
−12.35
−17.401
25.771
1
34.61
H


ATOM
2940
CZ
PHE
H
174
−11.594
−18.323
25.053
1
34.29
H


ATOM
2941
C
PHE
H
174
−12.099
−12.354
22.426
1
30.35
H


ATOM
2942
O
PHE
H
174
−11.035
−11.943
21.984
1
30.72
H


ATOM
2943
N
PRO
H
175
−13.155
−11.549
22.592
1
31.64
H


ATOM
2944
CD
PRO
H
175
−14.404
−11.865
23.303
1
34.54
H


ATOM
2945
CA
PRO
H
175
−13.101
−10.128
22.25
1
33.1
H


ATOM
2946
CB
PRO
H
175
−14.421
−9.585
22.808
1
32.79
H


ATOM
2947
CG
PRO
H
175
−14.757
−10.531
23.914
1
34.31
H


ATOM
2948
C
PRO
H
175
−11.876
−9.41
22.81
1
35.29
H


ATOM
2949
O
PRO
H
175
−11.34
−9.78
23.855
1
35.64
H


ATOM
2950
N
ALA
H
176
−11.435
−8.38
22.1
1
36.8
H


ATOM
2951
CA
ALA
H
176
−10.27
−7.617
22.513
1
39.38
H


ATOM
2952
CB
ALA
H
176
−9.719
−6.834
21.327
1
39.49
H


ATOM
2953
C
ALA
H
176
−10.599
−6.67
23.656
1
41.44
H


ATOM
2954
O
ALA
H
176
−11.761
−6.319
23.876
1
42.26
H


ATOM
2955
N
LEU
H
177
−9.567
−6.264
24.388
1
42.6
H


ATOM
2956
CA
LEU
H
177
−9.734
−5.341
25.501
1
42.99
H


ATOM
2957
CB
LEU
H
177
−9.694
−6.085
26.833
1
41.78
H


ATOM
2958
CG
LEU
H
177
−10.642
−7.268
26.989
1
41.02
H


ATOM
2959
CD1
LEU
H
177
−10.095
−8.463
26.214
1
44.32
H


ATOM
2960
CD2
LEU
H
177
−10.772
−7.618
28.456
1
40.99
H


ATOM
2961
C
LEU
H
177
−8.594
−4.342
25.449
1
44.35
H


ATOM
2962
O
LEU
H
177
−7.456
−4.713
25.149
1
46.44
H


ATOM
2963
N
LEU
H
178
−8.896
−3.076
25.724
1
43.97
H


ATOM
2964
CA
LEU
H
178
−7.867
−2.046
25.71
1
42.55
H


ATOM
2965
CB
LEU
H
178
−8.473
−0.658
25.521
1
39.68
H


ATOM
2966
CG
LEU
H
178
−8.545
−0.099
24.098
1
39.33
H


ATOM
2967
CD1
LEU
H
178
−8.813
1.405
24.17
1
34.97
H


ATOM
2968
CD2
LEU
H
178
−7.234
−0.36
23.359
1
36.38
H


ATOM
2969
C
LEU
H
178
−7.072
−2.057
26.996
1
43.27
H


ATOM
2970
O
LEU
H
178
−7.583
−2.414
28.056
1
43.59
H


ATOM
2971
N
GLN
H
179
−5.808
−1.673
26.891
1
45.59
H


ATOM
2972
CA
GLN
H
179
−4.927
−1.609
28.047
1
49.17
H


ATOM
2973
CB
GLN
H
179
−4.589
−3.013
28.561
1
52.14
H


ATOM
2974
CG
GLN
H
179
−3.741
−3.016
29.829
1
55.63
H


ATOM
2975
CD
GLN
H
179
−3.319
−4.418
30.251
1
60.07
H


ATOM
2976
OE1
GLN
H
179
−2.758
−5.176
29.457
1
62.32
H


ATOM
2977
NE2
GLN
H
179
−3.58
−4.765
31.508
1
61.27
H


ATOM
2978
C
GLN
H
179
−3.659
−0.887
27.624
1
48.5
H


ATOM
2979
O
GLN
H
179
−2.888
−1.396
26.807
1
48.4
H


ATOM
2980
N
SER
H
180
−3.468
0.31
28.174
1
46.95
H


ATOM
2981
CA
SER
H
180
−2.304
1.131
27.87
1
43.33
H


ATOM
2982
CB
SER
H
180
−1.022
0.353
28.152
1
43.86
H


ATOM
2983
OG
SER
H
180
−1.014
−0.124
29.484
1
45.65
H


ATOM
2984
C
SER
H
180
−2.322
1.582
26.417
1
41.61
H


ATOM
2985
O
SER
H
180
−1.272
1.737
25.791
1
41.93
H


ATOM
2986
N
GLY
H
183
−3.522
1.783
25.882
1
38.23
H


ATOM
2987
CA
GLY
H
183
−3.646
2.232
24.508
1
36.69
H


ATOM
2988
C
GLY
H
183
−3.568
1.119
23.483
1
36.11
H


ATOM
2989
O
GLY
H
183
−3.763
1.35
22.284
1
36.5
H


ATOM
2990
N
LEU
H
184
−3.29
−0.093
23.947
1
32.43
H


ATOM
2991
CA
LEU
H
184
−3.194
−1.221
23.049
1
29.54
H


ATOM
2992
CB
LEU
H
184
−1.813
−1.867
23.157
1
28.68
H


ATOM
2993
CG
LEU
H
184
−0.633
−0.982
22.757
1
26.45
H


ATOM
2994
CD1
LEU
H
184
0.644
−1.792
22.835
1
26.83
H


ATOM
2995
CD2
LEU
H
184
−0.835
−0.447
21.345
1
26.29
H


ATOM
2996
C
LEU
H
184
−4.267
−2.265
23.298
1
31.02
H


ATOM
2997
O
LEU
H
184
−4.671
−2.507
24.441
1
31.73
H


ATOM
2998
N
TYR
H
185
−4.718
−2.873
22.204
1
31.2
H


ATOM
2999
CA
TYR
H
185
−5.731
−3.919
22.228
1
32.06
H


ATOM
3000
CB
TYR
H
185
−6.533
−3.92
20.925
1
33.17
H


ATOM
3001
CG
TYR
H
185
−7.589
−2.858
20.843
1
35.48
H


ATOM
3002
CD1
TYR
H
185
−8.747
−2.944
21.613
1
37.25
H


ATOM
3003
CE1
TYR
H
185
−9.727
−1.958
21.552
1
39.1
H


ATOM
3004
CD2
TYR
H
185
−7.432
−1.759
20.005
1
38.27
H


ATOM
3005
CE2
TYR
H
185
−8.405
−0.765
19.936
1
41.84
H


ATOM
3006
CZ
TYR
H
185
−9.549
−0.872
20.714
1
41.51
H


ATOM
3007
OH
TYR
H
185
−10.507
0.113
20.661
1
46.97
H


ATOM
3008
C
TYR
H
185
−5.081
−5.286
22.376
1
32.82
H


ATOM
3009
O
TYR
H
185
−4.076
−5.591
21.725
1
32.31
H


ATOM
3010
N
THR
H
186
−5.644
−6.11
23.246
1
32.05
H


ATOM
3011
CA
THR
H
186
−5.126
−7.45
23.406
1
30.58
H


ATOM
3012
CB
THR
H
186
−4.459
−7.662
24.761
1
31.9
H


ATOM
3013
OG1
THR
H
186
−3.274
−6.857
24.833
1
35.66
H


ATOM
3014
CG2
THR
H
186
−4.074
−9.118
24.928
1
29.99
H


ATOM
3015
C
THR
H
186
−6.303
−8.378
23.24
1
29.64
H


ATOM
3016
O
THR
H
186
−7.43
−8.059
23.633
1
30.02
H


ATOM
3017
N
MET
H
187
−6.038
−9.526
22.639
1
27.85
H


ATOM
3018
CA
MET
H
187
−7.084
−10.484
22.37
1
26.53
H


ATOM
3019
CB
MET
H
187
−7.82
−10.03
21.103
1
26.47
H


ATOM
3020
CG
MET
H
187
−8.619
−11.082
20.394
1
31.53
H


ATOM
3021
SD
MET
H
187
−7.652
−11.96
19.158
1
33.24
H


ATOM
3022
CE
MET
H
187
−8.895
−13.055
18.545
1
29.37
H


ATOM
3023
C
MET
H
187
−6.481
−11.866
22.209
1
24.8
H


ATOM
3024
O
MET
H
187
−5.274
−12.007
22.067
1
27.57
H


ATOM
3025
N
SER
H
188
−7.319
−12.889
22.245
1
22.52
H


ATOM
3026
CA
SER
H
188
−6.837
−14.25
22.094
1
22.83
H


ATOM
3027
CB
SER
H
188
−6.438
−14.82
23.465
1
22.1
H


ATOM
3028
OG
SER
H
188
−7.537
−14.84
24.366
1
22.78
H


ATOM
3029
C
SER
H
188
−7.879
−15.159
21.439
1
22.3
H


ATOM
3030
O
SER
H
188
−9.04
−14.794
21.274
1
21.25
H


ATOM
3031
N
SER
H
189
−7.439
−16.348
21.057
1
22.45
H


ATOM
3032
CA
SER
H
189
−8.316
−17.333
20.459
1
20
H


ATOM
3033
CB
SER
H
189
−8.218
−17.278
18.934
1
20.62
H


ATOM
3034
OG
SER
H
189
−9.092
−18.218
18.328
1
25.43
H


ATOM
3035
C
SER
H
189
−7.834
−18.685
20.971
1
19.5
H


ATOM
3036
O
SER
H
189
−6.637
−18.891
21.163
1
15.41
H


ATOM
3037
N
SER
H
190
−8.764
−19.592
21.234
1
17.6
H


ATOM
3038
CA
SER
H
190
−8.38
−20.916
21.688
1
18.83
H


ATOM
3039
CB
SER
H
190
−8.93
−21.228
23.086
1
16.76
H


ATOM
3040
OG
SER
H
190
−10.303
−21.563
23.051
1
17.98
H


ATOM
3041
C
SER
H
190
−8.934
−21.909
20.692
1
20.96
H


ATOM
3042
O
SER
H
190
−9.926
−21.646
20.005
1
22.66
H


ATOM
3043
N
VAL
H
191
−8.269
−23.046
20.599
1
20.07
H


ATOM
3044
CA
VAL
H
191
−8.689
−24.09
19.696
1
18.23
H


ATOM
3045
CB
VAL
H
191
−7.883
−24.029
18.369
1
19.67
H


ATOM
3046
CG1
VAL
H
191
−6.392
−24.086
18.656
1
14.01
H


ATOM
3047
CG2
VAL
H
191
−8.307
−25.152
17.448
1
17.22
H


ATOM
3048
C
VAL
H
191
−8.427
−25.373
20.459
1
18.73
H


ATOM
3049
O
VAL
H
191
−7.378
−25.534
21.086
1
17.82
H


ATOM
3050
N
THR
H
192
−9.409
−26.263
20.449
1
19.74
H


ATOM
3051
CA
THR
H
192
−9.283
−27.526
21.155
1
18.94
H


ATOM
3052
CB
THR
H
192
−10.449
−27.697
22.148
1
17.25
H


ATOM
3053
OG1
THR
H
192
−10.294
−26.744
23.211
1
20.87
H


ATOM
3054
CG2
THR
H
192
−10.483
−29.096
22.727
1
12.72
H


ATOM
3055
C
THR
H
192
−9.273
−28.641
20.129
1
20.2
H


ATOM
3056
O
THR
H
192
−10.036
−28.601
19.155
1
23.51
H


ATOM
3057
N
VAL
H
193
−8.397
−29.622
20.335
1
16.09
H


ATOM
3058
CA
VAL
H
193
−8.285
−30.747
19.413
1
16.6
H


ATOM
3059
CB
VAL
H
193
−7.123
−30.529
18.427
1
15.63
H


ATOM
3060
CG1
VAL
H
193
−7.171
−29.118
17.879
1
16.41
H


ATOM
3061
CG2
VAL
H
193
−5.794
−30.806
19.113
1
15.42
H


ATOM
3062
C
VAL
H
193
−8.043
−32.065
20.148
1
18.34
H


ATOM
3063
O
VAL
H
193
−7.796
−32.077
21.357
1
19.21
H


ATOM
3064
N
PRO
H
194
−8.109
−33.197
19.426
1
19.37
H


ATOM
3065
CD
PRO
H
194
−8.563
−33.396
18.038
1
19.18
H


ATOM
3066
CA
PRO
H
194
−7.877
−34.486
20.084
1
22.24
H


ATOM
3067
CB
PRO
H
194
−8.116
−35.493
18.957
1
21.44
H


ATOM
3068
CG
PRO
H
194
−9.105
−34.8
18.077
1
18
H


ATOM
3069
C
PRO
H
194
−6.446
−34.565
20.644
1
24.6
H


ATOM
3070
O
PRO
H
194
−5.485
−34.253
19.943
1
24.2
H


ATOM
3071
N
SER
H
195
−6.318
−34.973
21.905
1
26.72
H


ATOM
3072
CA
SER
H
195
−5.014
−35.1
22.563
1
29.45
H


ATOM
3073
CB
SER
H
195
−5.193
−35.721
23.944
1
28.57
H


ATOM
3074
OG
SER
H
195
−6.008
−34.894
24.752
1
38.46
H


ATOM
3075
C
SER
H
195
−4.021
−35.943
21.765
1
29.09
H


ATOM
3076
O
SER
H
195
−2.827
−35.646
21.723
1
29.64
H


ATOM
3077
N
SER
H
196
−4.534
−37.001
21.151
1
29.83
H


ATOM
3078
CA
SER
H
196
−3.748
−37.916
20.336
1
31.64
H


ATOM
3079
CB
SER
H
196
−4.66
−39.007
19.786
1
31.29
H


ATOM
3080
OG
SER
H
196
−5.7
−38.419
19.014
1
31.39
H


ATOM
3081
C
SER
H
196
−3.084
−37.215
19.154
1
31.76
H


ATOM
3082
O
SER
H
196
−2.088
−37.693
18.618
1
33.34
H


ATOM
3083
N
THR
H
198
−3.64
−36.081
18.751
1
30.54
H


ATOM
3084
CA
THR
H
198
−3.12
−35.356
17.606
1
29.91
H


ATOM
3085
CB
THR
H
198
−4.267
−34.692
16.837
1
29.73
H


ATOM
3086
OG1
THR
H
198
−4.786
−33.597
17.603
1
27.57
H


ATOM
3087
CG2
THR
H
198
−5.376
−35.712
16.58
1
25.77
H


ATOM
3088
C
THR
H
198
−2.06
−34.305
17.906
1
29.71
H


ATOM
3089
O
THR
H
198
−1.444
−33.76
16.982
1
27.43
H


ATOM
3090
N
TRP
H
199
−1.84
−34.008
19.182
1
29.35
H


ATOM
3091
CA
TRP
H
199
−0.822
−33.021
19.524
1
28.32
H


ATOM
3092
CB
TRP
H
199
−1.438
−31.832
20.252
1
28.31
H


ATOM
3093
CG
TRP
H
199
−0.739
−30.566
19.9
1
29.39
H


ATOM
3094
CD2
TRP
H
199
0.129
−29.807
20.742
1
30.09
H


ATOM
3095
CE2
TRP
H
199
0.648
−28.746
19.96
1
30.58
H


ATOM
3096
CE3
TRP
H
199
0.526
−29.919
22.08
1
29.79
H


ATOM
3097
CD1
TRP
H
199
−0.722
−29.953
18.678
1
31.51
H


ATOM
3098
NE1
TRP
H
199
0.112
−28.858
18.705
1
30.06
H


ATOM
3099
CZ2
TRP
H
199
1.542
−27.804
20.474
1
29.69
H


ATOM
3100
CZ3
TRP
H
199
1.416
−28.984
22.591
1
31.66
H


ATOM
3101
CH2
TRP
H
199
1.914
−27.938
21.786
1
30.85
H


ATOM
3102
C
TRP
H
199
0.296
−33.622
20.368
1
26.48
H


ATOM
3103
O
TRP
H
199
0.057
−34.481
21.208
1
27.32
H


ATOM
3104
N
PRO
H
200
1.539
−33.157
20.166
1
26.79
H


ATOM
3105
CD
PRO
H
200
2.675
−33.526
21.03
1
23.84
H


ATOM
3106
CA
PRO
H
200
1.947
−32.117
19.215
1
25.58
H


ATOM
3107
CB
PRO
H
200
3.113
−31.471
19.924
1
25.21
H


ATOM
3108
CG
PRO
H
200
3.823
−32.689
20.449
1
23.8
H


ATOM
3109
C
PRO
H
200
2.372
−32.635
17.848
1
25.26
H


ATOM
3110
O
PRO
H
200
3.189
−32.005
17.186
1
27.47
H


ATOM
3111
N
SER
H
202
1.84
−33.775
17.427
1
25.77
H


ATOM
3112
CA
SER
H
202
2.216
−34.328
16.132
1
25.92
H


ATOM
3113
CB
SER
H
202
1.859
−35.813
16.058
1
25.33
H


ATOM
3114
OG
SER
H
202
0.458
−36.001
16.15
1
35.16
H


ATOM
3115
C
SER
H
202
1.585
−33.581
14.956
1
25.44
H


ATOM
3116
O
SER
H
202
2.186
−33.498
13.89
1
25.63
H


ATOM
3117
N
GLN
H
203
0.389
−33.027
15.134
1
26.68
H


ATOM
3118
CA
GLN
H
203
−0.239
−32.307
14.027
1
29.32
H


ATOM
3119
CB
GLN
H
203
−1.771
−32.341
14.113
1
27.47
H


ATOM
3120
CG
GLN
H
203
−2.373
−31.332
15.077
1
31.13
H


ATOM
3121
CD
GLN
H
203
−3.861
−31.132
14.852
1
33.58
H


ATOM
3122
OE1
GLN
H
203
−4.276
−30.566
13.84
1
34.02
H


ATOM
3123
NE2
GLN
H
203
−4.672
−31.603
15.794
1
33.5
H


ATOM
3124
C
GLN
H
203
0.211
−30.859
13.974
1
29.93
H


ATOM
3125
O
GLN
H
203
0.769
−30.321
14.934
1
33.44
H


ATOM
3126
N
THR
H
204
−0.045
−30.227
12.837
1
29.05
H


ATOM
3127
CA
THR
H
204
0.319
−28.84
12.647
1
25.77
H


ATOM
3128
CB
THR
H
204
0.916
−28.636
11.266
1
24.61
H


ATOM
3129
OG1
THR
H
204
2.154
−29.353
11.192
1
21.97
H


ATOM
3130
CG2
THR
H
204
1.149
−27.163
11.001
1
22.09
H


ATOM
3131
C
THR
H
204
−0.898
−27.952
12.825
1
26.63
H


ATOM
3132
O
THR
H
204
−1.833
−27.98
12.028
1
27.24
H


ATOM
3133
N
VAL
H
205
−0.878
−27.173
13.896
1
28.23
H


ATOM
3134
CA
VAL
H
205
−1.966
−26.266
14.216
1
26.73
H


ATOM
3135
CB
VAL
H
205
−2.541
−26.565
15.603
1
26.06
H


ATOM
3136
CG1
VAL
H
205
−3.438
−25.432
16.049
1
26.94
H


ATOM
3137
CG2
VAL
H
205
−3.309
−27.876
15.568
1
26.49
H


ATOM
3138
C
VAL
H
205
−1.421
−24.858
14.222
1
28.23
H


ATOM
3139
O
VAL
H
205
−0.537
−24.531
15.016
1
27.35
H


ATOM
3140
N
THR
H
206
−1.938
−24.02
13.333
1
27.87
H


ATOM
3141
CA
THR
H
206
−1.467
−22.651
13.278
1
29.33
H


ATOM
3142
CB
THR
H
206
−0.547
−22.419
12.057
1
30.95
H


ATOM
3143
OG1
THR
H
206
−1.345
−22.243
10.883
1
33.35
H


ATOM
3144
CG2
THR
H
206
0.373
−23.613
11.853
1
30.47
H


ATOM
3145
C
THR
H
206
−2.606
−21.647
13.225
1
28.51
H


ATOM
3146
O
THR
H
206
−3.655
−21.9
12.629
1
28.96
H


ATOM
3147
N
CYS
H
208
−2.384
−20.511
13.876
1
27.16
H


ATOM
3148
CA
CYS
H
208
−3.348
−19.43
13.897
1
26.06
H


ATOM
3149
C
CYS
H
208
−2.754
−18.323
13.031
1
24.84
H


ATOM
3150
O
CYS
H
208
−1.66
−17.827
13.296
1
24.56
H


ATOM
3151
CB
CYS
H
208
−3.545
−18.906
15.327
1
28.28
H


ATOM
3152
SG
CYS
H
208
−2.094
−18.015
15.991
1
30.17
H


ATOM
3153
N
SER
H
209
−3.452
−17.963
11.969
1
23.05
H


ATOM
3154
CA
SER
H
209
−2.973
−16.892
11.117
1
22.76
H


ATOM
3155
CB
SER
H
209
−3.253
−17.21
9.649
1
21.71
H


ATOM
3156
OG
SER
H
209
−4.486
−17.892
9.51
1
30.48
H


ATOM
3157
C
SER
H
209
−3.736
−15.669
11.597
1
21.89
H


ATOM
3158
O
SER
H
209
−4.966
−15.658
11.618
1
25.21
H


ATOM
3159
N
VAL
H
210
−2.993
−14.658
12.02
1
18.09
H


ATOM
3160
CA
VAL
H
210
−3.577
−13.448
12.558
1
16.35
H


ATOM
3161
CB
VAL
H
210
−2.85
−13.025
13.859
1
14.54
H


ATOM
3162
CG1
VAL
H
210
−3.513
−11.811
14.466
1
16.17
H


ATOM
3163
CG2
VAL
H
210
−2.858
−14.167
14.85
1
12.31
H


ATOM
3164
C
VAL
H
210
−3.516
−12.302
11.568
1
21.67
H


ATOM
3165
O
VAL
H
210
−2.441
−11.779
11.266
1
20.4
H


ATOM
3166
N
ALA
H
211
−4.686
−11.91
11.076
1
25.15
H


ATOM
3167
CA
ALA
H
211
−4.793
−10.819
10.12
1
28.64
H


ATOM
3168
CB
ALA
H
211
−5.837
−11.159
9.06
1
27.27
H


ATOM
3169
C
ALA
H
211
−5.151
−9.494
10.799
1
30.29
H


ATOM
3170
O
ALA
H
211
−6.108
−9.408
11.579
1
31.11
H


ATOM
3171
N
HIS
H
212
−4.367
−8.465
10.496
1
30.86
H


ATOM
3172
CA
HIS
H
212
−4.589
−7.131
11.034
1
31.54
H


ATOM
3173
CB
HIS
H
212
−3.385
−6.699
11.87
1
32.43
H


ATOM
3174
CG
HIS
H
212
−3.613
−5.451
12.667
1
32.79
H


ATOM
3175
CD2
HIS
H
212
−4.58
−5.126
13.556
1
33.22
H


ATOM
3176
ND1
HIS
H
212
−2.766
−4.366
12.608
1
32.09
H


ATOM
3177
CE1
HIS
H
212
−3.2
−3.425
13.426
1
32.9
H


ATOM
3178
NE2
HIS
H
212
−4.3
−3.861
14.014
1
33.97
H


ATOM
3179
C
HIS
H
212
−4.73
−6.234
9.8
1
33.24
H


ATOM
3180
O
HIS
H
212
−3.747
−5.663
9.322
1
32.73
H


ATOM
3181
N
PRO
H
213
−5.956
−6.115
9.256
1
32.76
H


ATOM
3182
CD
PRO
H
213
−7.228
−6.729
9.681
1
30
H


ATOM
3183
CA
PRO
H
213
−6.157
−5.279
8.069
1
32.17
H


ATOM
3184
CB
PRO
H
213
−7.681
−5.213
7.952
1
31.98
H


ATOM
3185
CG
PRO
H
213
−8.096
−6.573
8.438
1
29.41
H


ATOM
3186
C
PRO
H
213
−5.505
−3.904
8.157
1
31.23
H


ATOM
3187
O
PRO
H
213
−4.78
−3.495
7.246
1
30.71
H


ATOM
3188
N
ALA
H
214
−5.749
−3.203
9.259
1
30.34
H


ATOM
3189
CA
ALA
H
214
−5.189
−1.869
9.451
1
31.28
H


ATOM
3190
CB
ALA
H
214
−5.462
−1.394
10.867
1
33.75
H


ATOM
3191
C
ALA
H
214
−3.687
−1.771
9.148
1
32.3
H


ATOM
3192
O
ALA
H
214
−3.22
−0.736
8.673
1
33.96
H


ATOM
3193
N
SER
H
215
−2.926
−2.829
9.417
1
31.08
H


ATOM
3194
CA
SER
H
215
−1.491
−2.783
9.153
1
30.2
H


ATOM
3195
CB
SER
H
215
−0.699
−3.272
10.375
1
33.35
H


ATOM
3196
OG
SER
H
215
−0.753
−4.679
10.516
1
34.86
H


ATOM
3197
C
SER
H
215
−1.105
−3.596
7.922
1
30.55
H


ATOM
3198
O
SER
H
215
0.078
−3.775
7.627
1
30.68
H


ATOM
3199
N
SER
H
216
−2.111
−4.087
7.204
1
30.46
H


ATOM
3200
CA
SER
H
216
−1.88
−4.871
5.994
1
30.3
H


ATOM
3201
CB
SER
H
216
−1.207
−3.986
4.943
1
28.9
H


ATOM
3202
OG
SER
H
216
−1.949
−2.791
4.763
1
31.14
H


ATOM
3203
C
SER
H
216
−1.026
−6.113
6.266
1
29.8
H


ATOM
3204
O
SER
H
216
−0.324
−6.602
5.382
1
28.07
H


ATOM
3205
N
THR
H
217
−1.106
−6.629
7.489
1
30.23
H


ATOM
3206
CA
THR
H
217
−0.325
−7.793
7.87
1
30.99
H


ATOM
3207
CB
THR
H
217
0.596
−7.479
9.05
1
32.81
H


ATOM
3208
OG1
THR
H
217
−0.189
−7.044
10.169
1
31.41
H


ATOM
3209
CG2
THR
H
217
1.585
−6.411
8.666
1
34.03
H


ATOM
3210
C
THR
H
217
−1.117
−9.03
8.251
1
30.77
H


ATOM
3211
O
THR
H
217
−2.311
−8.98
8.555
1
29.94
H


ATOM
3212
N
THR
H
218
−0.409
−10.146
8.241
1
29.71
H


ATOM
3213
CA
THR
H
218
−0.981
−11.42
8.592
1
31.98
H


ATOM
3214
CB
THR
H
218
−1.667
−12.072
7.376
1
33.33
H


ATOM
3215
OG1
THR
H
218
−2.872
−11.354
7.078
1
35.24
H


ATOM
3216
CG2
THR
H
218
−2.006
−13.533
7.657
1
33.67
H


ATOM
3217
C
THR
H
218
0.164
−12.276
9.085
1
31.47
H


ATOM
3218
O
THR
H
218
1.096
−12.571
8.339
1
31.87
H


ATOM
3219
N
VAL
H
219
0.095
−12.656
10.357
1
32.84
H


ATOM
3220
CA
VAL
H
219
1.128
−13.477
10.964
1
34.38
H


ATOM
3221
CB
VAL
H
219
1.57
−12.918
12.312
1
34.49
H


ATOM
3222
CG1
VAL
H
219
2.7
−13.776
12.861
1
37.59
H


ATOM
3223
CG2
VAL
H
219
2.022
−11.469
12.157
1
34.89
H


ATOM
3224
C
VAL
H
219
0.679
−14.909
11.188
1
35.93
H


ATOM
3225
O
VAL
H
219
−0.377
−15.166
11.768
1
34.39
H


ATOM
3226
N
ASP
H
220
1.501
−15.842
10.73
1
36.44
H


ATOM
3227
CA
ASP
H
220
1.2
−17.25
10.881
1
37.22
H


ATOM
3228
CB
ASP
H
220
1.545
−17.988
9.595
1
41.17
H


ATOM
3229
CG
ASP
H
220
0.624
−17.619
8.461
1
45.72
H


ATOM
3230
OD1
ASP
H
220
−0.594
−17.841
8.61
1
50.06
H


ATOM
3231
OD2
ASP
H
220
1.106
−17.105
7.428
1
50.88
H


ATOM
3232
C
ASP
H
220
1.968
−17.847
12.047
1
35.86
H


ATOM
3233
O
ASP
H
220
3.196
−17.83
12.066
1
38.85
H


ATOM
3234
N
LYS
H
221
1.242
−18.369
13.027
1
32.81
H


ATOM
3235
CA
LYS
H
221
1.885
−18.972
14.18
1
30.93
H


ATOM
3236
CB
LYS
H
221
1.395
−18.329
15.479
1
31.24
H


ATOM
3237
CG
LYS
H
221
2.43
−17.481
16.167
1
31.26
H


ATOM
3238
CD
LYS
H
221
3.737
−18.231
16.361
1
28.08
H


ATOM
3239
CE
LYS
H
221
4.831
−17.249
16.725
1
26.73
H


ATOM
3240
NZ
LYS
H
221
6.184
−17.853
16.668
1
32.05
H


ATOM
3241
C
LYS
H
221
1.65
−20.467
14.264
1
29.05
H


ATOM
3242
O
LYS
H
221
0.533
−20.919
14.507
1
22.27
H


ATOM
3243
N
LYS
H
222
2.709
−21.235
14.053
1
29.9
H


ATOM
3244
CA
LYS
H
222
2.599
−22.675
14.166
1
34.28
H


ATOM
3245
CB
LYS
H
222
3.69
−23.376
13.35
1
34.48
H


ATOM
3246
CG
LYS
H
222
3.831
−24.868
13.64
1
34.6
H


ATOM
3247
CD
LYS
H
222
4.998
−25.452
12.85
1
40.57
H


ATOM
3248
CE
LYS
H
222
5.329
−26.886
13.264
1
43.98
H


ATOM
3249
NZ
LYS
H
222
4.198
−27.839
13.083
1
44.62
H


ATOM
3250
C
LYS
H
222
2.809
−22.927
15.647
1
35.44
H


ATOM
3251
O
LYS
H
222
3.762
−22.42
16.238
1
36.24
H


ATOM
3252
N
LEU
H
223
1.911
−23.682
16.261
1
37.71
H


ATOM
3253
CA
LEU
H
223
2.06
−23.959
17.676
1
41.2
H


ATOM
3254
CB
LEU
H
223
0.733
−24.436
18.266
1
40.43
H


ATOM
3255
CG
LEU
H
223
−0.44
−23.472
18.046
1
43.19
H


ATOM
3256
CD1
LEU
H
223
−1.594
−23.885
18.938
1
44.63
H


ATOM
3257
CD2
LEU
H
223
−0.034
−22.042
18.358
1
40.69
H


ATOM
3258
C
LEU
H
223
3.158
−24.994
17.9
1
44.39
H


ATOM
3259
O
LEU
H
223
3.28
−25.975
17.155
1
44.48
H


ATOM
3260
N
GLU
H
226
3.97
−24.758
18.923
1
47.06
H


ATOM
3261
CA
GLU
H
226
5.063
−25.659
19.25
1
50.27
H


ATOM
3262
CB
GLU
H
226
6.405
−24.953
19.033
1
50.04
H


ATOM
3263
CG
GLU
H
226
6.605
−24.412
17.621
1
50.6
H


ATOM
3264
CD
GLU
H
226
6.919
−25.495
16.604
1
48.9
H


ATOM
3265
OE1
GLU
H
226
6.256
−26.556
16.624
1
47.3
H


ATOM
3266
OE2
GLU
H
226
7.826
−25.273
15.775
1
47.59
H


ATOM
3267
C
GLU
H
226
4.933
−26.08
20.704
1
52.54
H


ATOM
3268
O
GLU
H
226
4.414
−25.326
21.524
1
53.07
H


ATOM
3269
N
PRO
H
227
5.384
−27.301
21.04
1
55.66
H


ATOM
3270
CD
PRO
H
227
5.853
−28.358
20.126
1
55.34
H


ATOM
3271
CA
PRO
H
227
5.311
−27.797
22.421
1
57.84
H


ATOM
3272
CB
PRO
H
227
5.907
−29.198
22.314
1
56.01
H


ATOM
3273
CG
PRO
H
227
5.565
−29.603
20.913
1
55.76
H


ATOM
3274
C
PRO
H
227
6.133
−26.892
23.343
1
60.3
H


ATOM
3275
O
PRO
H
227
7.332
−26.713
23.13
1
61.08
H


ATOM
3276
N
SER
H
228
5.492
−26.322
24.361
1
63.5
H


ATOM
3277
CA
SER
H
228
6.189
−25.428
25.285
1
66.55
H


ATOM
3278
CB
SER
H
228
5.224
−24.356
25.818
1
67.37
H


ATOM
3279
OG
SER
H
228
4.06
−24.926
26.387
1
68.18
H


ATOM
3280
C
SER
H
228
6.867
−26.147
26.449
1
67.17
H


ATOM
3281
O
SER
H
228
8.102
−25.991
26.592
1
68.28
H


ATOM
3282
OXT
SER
H
228
6.165
−26.852
27.204
1
68.18
H


ATOM
3283
CB
THR
A
300
−39.993
20.06
−5.025
1
63.64
A


ATOM
3284
OG1
THR
A
300
−38.637
19.597
−5.063
1
64.49
A


ATOM
3285
CG2
THR
A
300
−40.076
21.348
−4.208
1
63.52
A


ATOM
3286
C
THR
A
300
−41.057
17.834
−5.415
1
60.23
A


ATOM
3287
O
THR
A
300
−42.138
17.255
−5.509
1
62.42
A


ATOM
3288
N
THR
A
300
−40.313
18.493
−3.114
1
60.6
A


ATOM
3289
CA
THR
A
300
−40.883
18.971
−4.405
1
61.67
A


ATOM
3290
N
THR
A
301
−40.004
17.514
−6.166
1
57.02
A


ATOM
3291
CA
THR
A
301
−40.074
16.445
−7.171
1
53.86
A


ATOM
3292
CB
THR
A
301
−40.912
16.886
−8.388
1
54.85
A


ATOM
3293
OG1
THR
A
301
−42.277
17.061
−7.988
1
55.11
A


ATOM
3294
CG2
THR
A
301
−40.839
15.846
−9.496
1
54.22
A


ATOM
3295
C
THR
A
301
−38.699
15.997
−7.668
1
50.39
A


ATOM
3296
O
THR
A
301
−37.799
16.816
−7.854
1
52.5
A


ATOM
3297
N
TYR
A
302
−38.548
14.696
−7.903
1
44.26
A


ATOM
3298
CA
TYR
A
302
−37.273
14.149
−8.355
1
38.64
A


ATOM
3299
CB
TYR
A
302
−36.757
13.132
−7.334
1
38.4
A


ATOM
3300
CG
TYR
A
302
−36.54
13.739
−5.965
1
37.35
A


ATOM
3301
CD1
TYR
A
302
−35.445
14.56
−5.714
1
36.62
A


ATOM
3302
CE1
TYR
A
302
−35.276
15.173
−4.478
1
38.19
A


ATOM
3303
CD2
TYR
A
302
−37.465
13.541
−4.941
1
36.87
A


ATOM
3304
CE2
TYR
A
302
−37.31
14.146
−3.705
1
37.5
A


ATOM
3305
CZ
TYR
A
302
−36.213
14.965
−3.474
1
40.16
A


ATOM
3306
OH
TYR
A
302
−36.055
15.577
−2.244
1
39.05
A


ATOM
3307
C
TYR
A
302
−37.337
13.516
−9.734
1
35.23
A


ATOM
3308
O
TYR
A
302
−38.221
13.83
−10.527
1
36.08
A


ATOM
3309
N
GLY
A
303
−36.389
12.628
−10.014
1
31.15
A


ATOM
3310
CA
GLY
A
303
−36.338
11.974
−11.306
1
29
A


ATOM
3311
C
GLY
A
303
−37.35
10.864
−11.496
1
27.36
A


ATOM
3312
O
GLY
A
303
−38.487
10.96
−11.054
1
27.78
A


ATOM
3313
N
VAL
A
304
−36.928
9.805
−12.176
1
28.37
A


ATOM
3314
CA
VAL
A
304
−37.783
8.653
−12.439
1
25.45
A


ATOM
3315
CB
VAL
A
304
−37.355
7.941
−13.743
1
23.02
A


ATOM
3316
CG1
VAL
A
304
−38.345
6.841
−14.08
1
22.44
A


ATOM
3317
CG2
VAL
A
304
−37.266
8.937
−14.88
1
19.78
A


ATOM
3318
C
VAL
A
304
−37.644
7.674
−11.275
1
25.59
A


ATOM
3319
O
VAL
A
304
−36.532
7.412
−10.818
1
27.5
A


ATOM
3320
N
CYS
A
305
−38.761
7.143
−10.786
1
24.55
A


ATOM
3321
CA
CYS
A
305
−38.714
6.192
−9.675
1
23.42
A


ATOM
3322
C
CYS
A
305
−37.698
5.08
−9.971
1
24.43
A


ATOM
3323
O
CYS
A
305
−37.74
4.453
−11.031
1
23.32
A


ATOM
3324
CB
CYS
A
305
−40.096
5.591
−9.433
1
23.21
A


ATOM
3325
SG
CYS
A
305
−41.409
6.808
−9.079
1
22.86
A


ATOM
3326
N
SER
A
306
−36.791
4.848
−9.022
1
25.15
A


ATOM
3327
CA
SER
A
306
−35.723
3.853
−9.15
1
22.43
A


ATOM
3328
CB
SER
A
306
−34.448
4.401
−8.509
1
22.2
A


ATOM
3329
OG
SER
A
306
−34.647
4.636
−7.124
1
17.99
A


ATOM
3330
C
SER
A
306
−36.032
2.483
−8.541
1
21.94
A


ATOM
3331
O
SER
A
306
−35.336
1.506
−8.814
1
20.23
A


ATOM
3332
N
LYS
A
307
−37.061
2.405
−7.708
1
20.25
A


ATOM
3333
CA
LYS
A
307
−37.415
1.128
−7.1
1
20.29
A


ATOM
3334
CB
LYS
A
307
−37.89
1.336
−5.661
1
17.83
A


ATOM
3335
CG
LYS
A
307
−36.843
1.928
−4.742
1
18.48
A


ATOM
3336
CD
LYS
A
307
−35.585
1.08
−4.7
1
20.11
A


ATOM
3337
CE
LYS
A
307
−34.685
1.512
−3.562
1
23.13
A


ATOM
3338
NZ
LYS
A
307
−33.554
0.58
−3.379
1
22.66
A


ATOM
3339
C
LYS
A
307
−38.496
0.398
−7.903
1
19.89
A


ATOM
3340
O
LYS
A
307
−39.182
0.984
−8.734
1
19.77
A


ATOM
3341
N
ALA
A
308
−38.637
−0.893
−7.655
1
19.8
A


ATOM
3342
CA
ALA
A
308
−39.633
−1.683
−8.349
1
22.06
A


ATOM
3343
CB
ALA
A
308
−39.309
−3.16
−8.199
1
17.28
A


ATOM
3344
C
ALA
A
308
−41.046
−1.395
−7.827
1
23.71
A


ATOM
3345
O
ALA
A
308
−41.231
−0.934
−6.697
1
22.91
A


ATOM
3346
N
PHE
A
309
−42.035
−1.658
−8.673
1
25.12
A


ATOM
3347
CA
PHE
A
309
−43.428
−1.468
−8.317
1
26.85
A


ATOM
3348
CB
PHE
A
309
−44.183
−0.705
−9.407
1
26.25
A


ATOM
3349
CG
PHE
A
309
−43.804
0.741
−9.52
1
29.27
A


ATOM
3350
CD1
PHE
A
309
−42.56
1.116
−10.019
1
28.25
A


ATOM
3351
CD2
PHE
A
309
−44.701
1.736
−9.13
1
28.22
A


ATOM
3352
CE1
PHE
A
309
−42.212
2.463
−10.13
1
27.47
A


ATOM
3353
CE2
PHE
A
309
−44.365
3.084
−9.236
1
27.52
A


ATOM
3354
CZ
PHE
A
309
−43.118
3.449
−9.738
1
28.7
A


ATOM
3355
C
PHE
A
309
−44.051
−2.845
−8.193
1
29.88
A


ATOM
3356
O
PHE
A
309
−43.408
−3.864
−8.472
1
26.84
A


ATOM
3357
N
LYS
A
310
−45.312
−2.869
−7.779
1
30.73
A


ATOM
3358
CA
LYS
A
310
−46.035
−4.116
−7.651
1
31.75
A


ATOM
3359
CB
LYS
A
310
−45.987
−4.617
−6.202
1
35.18
A


ATOM
3360
CG
LYS
A
310
−46.277
−6.11
−6.067
1
40.9
A


ATOM
3361
CD
LYS
A
310
−46.153
−6.619
−4.63
1
44.59
A


ATOM
3362
CE
LYS
A
310
−46.262
−8.154
−4.58
1
46.43
A


ATOM
3363
NZ
LYS
A
310
−46.255
−8.719
−3.193
1
49.07
A


ATOM
3364
C
LYS
A
310
−47.478
−3.881
−8.095
1
32.4
A


ATOM
3365
O
LYS
A
310
−48.092
−2.864
−7.755
1
29.67
A


ATOM
3366
N
PHE
A
311
−48
−4.811
−8.886
1
33.4
A


ATOM
3367
CA
PHE
A
311
−49.372
−4.725
−9.367
1
37.04
A


ATOM
3368
CB
PHE
A
311
−49.557
−5.616
−10.601
1
36.39
A


ATOM
3369
CG
PHE
A
311
−49.018
−5.023
−11.877
1
35.69
A


ATOM
3370
CD1
PHE
A
311
−49.786
−4.137
−12.627
1
37.17
A


ATOM
3371
CD2
PHE
A
311
−47.75
−5.356
−12.333
1
35.82
A


ATOM
3372
CE1
PHE
A
311
−49.298
−3.594
−13.817
1
37.02
A


ATOM
3373
CE2
PHE
A
311
−47.251
−4.817
−13.519
1
36.63
A


ATOM
3374
CZ
PHE
A
311
−48.027
−3.936
−14.262
1
35.98
A


ATOM
3375
C
PHE
A
311
−50.335
−5.174
−8.268
1
38.96
A


ATOM
3376
O
PHE
A
311
−50.138
−6.215
−7.649
1
40.29
A


ATOM
3377
N
LEU
A
312
−51.37
−4.38
−8.024
1
41.96
A


ATOM
3378
CA
LEU
A
312
−52.374
−4.713
−7.017
1
45.5
A


ATOM
3379
CB
LEU
A
312
−52.713
−3.473
−6.19
1
44.96
A


ATOM
3380
CG
LEU
A
312
−51.477
−2.789
−5.61
1
44.34
A


ATOM
3381
CD1
LEU
A
312
−51.845
−1.424
−5.073
1
44.91
A


ATOM
3382
CD2
LEU
A
312
−50.873
−3.67
−4.529
1
44.77
A


ATOM
3383
C
LEU
A
312
−53.624
−5.225
−7.737
1
47.79
A


ATOM
3384
O
LEU
A
312
−54.346
−4.463
−8.391
1
50.15
A


ATOM
3385
N
GLY
A
313
−53.875
−6.52
−7.618
1
49.8
A


ATOM
3386
CA
GLY
A
313
−55.024
−7.105
−8.283
1
51.57
A


ATOM
3387
C
GLY
A
313
−54.628
−7.535
−9.681
1
52.43
A


ATOM
3388
O
GLY
A
313
−53.66
−8.277
−9.869
1
54.68
A


ATOM
3389
N
THR
A
314
−55.374
−7.059
−10.669
1
53.04
A


ATOM
3390
CA
THR
A
314
−55.105
−7.385
−12.064
1
51.49
A


ATOM
3391
CB
THR
A
314
−55.682
−8.762
−12.432
1
52.44
A


ATOM
3392
OG1
THR
A
314
−54.968
−9.78
−11.719
1
54.54
A


ATOM
3393
CG2
THR
A
314
−55.571
−9.009
−13.932
1
53.08
A


ATOM
3394
C
THR
A
314
−55.742
−6.33
−12.948
1
48.79
A


ATOM
3395
O
THR
A
314
−56.896
−5.956
−12.741
1
48.63
A


ATOM
3396
N
PRO
A
315
−54.998
−5.831
−13.946
1
47.38
A


ATOM
3397
CD
PRO
A
315
−53.645
−6.226
−14.377
1
47.03
A


ATOM
3398
CA
PRO
A
315
−55.551
−4.81
−14.834
1
46.88
A


ATOM
3399
CB
PRO
A
315
−54.616
−4.863
−16.032
1
46.34
A


ATOM
3400
CG
PRO
A
315
−53.296
−5.143
−15.381
1
47.58
A


ATOM
3401
C
PRO
A
315
−57.007
−5.095
−15.189
1
47.46
A


ATOM
3402
O
PRO
A
315
−57.383
−6.225
−15.525
1
44.71
A


ATOM
3403
N
ALA
A
316
−57.824
−4.055
−15.091
1
47.6
A


ATOM
3404
CA
ALA
A
316
−59.239
−4.173
−15.372
1
46.94
A


ATOM
3405
CB
ALA
A
316
−60.039
−3.598
−14.206
1
47.42
A


ATOM
3406
C
ALA
A
316
−59.636
−3.482
−16.669
1
46.66
A


ATOM
3407
O
ALA
A
316
−59.104
−2.431
−17.023
1
43.77
A


ATOM
3408
N
ASP
A
317
−60.582
−4.095
−17.371
1
48.84
A


ATOM
3409
CA
ASP
A
317
−61.097
−3.555
−18.618
1
49.79
A


ATOM
3410
CB
ASP
A
317
−61.752
−4.674
−19.426
1
51.92
A


ATOM
3411
CG
ASP
A
317
−61.941
−4.306
−20.885
1
55.75
A


ATOM
3412
OD1
ASP
A
317
−62.581
−3.266
−21.163
1
55.59
A


ATOM
3413
OD2
ASP
A
317
−61.444
−5.06
−21.753
1
56.72
A


ATOM
3414
C
ASP
A
317
−62.139
−2.5
−18.241
1
49.14
A


ATOM
3415
O
ASP
A
317
−63.011
−2.765
−17.419
1
48.28
A


ATOM
3416
N
THR
A
318
−62.046
−1.308
−18.828
1
49.13
A


ATOM
3417
CA
THR
A
318
−62.994
−0.23
−18.522
1
48.54
A


ATOM
3418
CB
THR
A
318
−62.342
1.172
−18.637
1
45.08
A


ATOM
3419
OG1
THR
A
318
−61.91
1.391
−19.983
1
42.12
A


ATOM
3420
CG2
THR
A
318
−61.163
1.292
−17.712
1
42.82
A


ATOM
3421
C
THR
A
318
−64.196
−0.234
−19.456
1
50.01
A


ATOM
3422
O
THR
A
318
−65.061
0.64
−19.367
1
49.96
A


ATOM
3423
N
GLY
A
319
−64.247
−1.21
−20.355
1
50.29
A


ATOM
3424
CA
GLY
A
319
−65.35
−1.269
−21.293
1
50.96
A


ATOM
3425
C
GLY
A
319
−65.344
−0.036
−22.174
1
51.2
A


ATOM
3426
O
GLY
A
319
−66.272
0.187
−22.95
1
53.64
A


ATOM
3427
N
HIS
A
320
−64.291
0.767
−22.05
1
51.62
A


ATOM
3428
CA
HIS
A
320
−64.149
1.985
−22.837
1
51.32
A


ATOM
3429
CB
HIS
A
320
−64.009
3.204
−21.921
1
54.93
A


ATOM
3430
CG
HIS
A
320
−65.103
3.331
−20.906
1
59.78
A


ATOM
3431
CD2
HIS
A
320
−65.057
3.473
−19.559
1
61.42
A


ATOM
3432
ND1
HIS
A
320
−66.439
3.326
−21.244
1
61.23
A


ATOM
3433
CE1
HIS
A
320
−67.169
3.457
−20.15
1
62.96
A


ATOM
3434
NE2
HIS
A
320
−66.355
3.548
−19.114
1
63.22
A


ATOM
3435
C
HIS
A
320
−62.923
1.898
−23.735
1
49.8
A


ATOM
3436
O
HIS
A
320
−62.558
2.87
−24.387
1
50.31
A


ATOM
3437
N
GLY
A
321
−62.284
0.733
−23.763
1
48.67
A


ATOM
3438
CA
GLY
A
321
−61.103
0.563
−24.593
1
46.45
A


ATOM
3439
C
GLY
A
321
−59.819
0.795
−23.816
1
46.22
A


ATOM
3440
O
GLY
A
321
−58.713
0.692
−24.355
1
46.09
A


ATOM
3441
N
THR
A
322
−59.967
1.113
−22.536
1
43.6
A


ATOM
3442
CA
THR
A
322
−58.816
1.353
−21.681
1
39.55
A


ATOM
3443
CB
THR
A
322
−58.844
2.763
−21.092
1
38.16
A


ATOM
3444
OG1
THR
A
322
−59.945
2.875
−20.182
1
37.52
A


ATOM
3445
CG2
THR
A
322
−58.991
3.792
−22.19
1
37.16
A


ATOM
3446
C
THR
A
322
−58.813
0.373
−20.521
1
37.54
A


ATOM
3447
O
THR
A
322
−59.836
−0.245
−20.21
1
35.62
A


ATOM
3448
N
VAL
A
323
−57.656
0.226
−19.889
1
35.39
A


ATOM
3449
CA
VAL
A
323
−57.534
−0.655
−18.738
1
34.7
A


ATOM
3450
CB
VAL
A
323
−56.494
−1.772
−18.978
1
32.69
A


ATOM
3451
CG1
VAL
A
323
−56.696
−2.357
−20.357
1
31.9
A


ATOM
3452
CG2
VAL
A
323
−55.082
−1.231
−18.827
1
31.65
A


ATOM
3453
C
VAL
A
323
−57.101
0.189
−17.548
1
34.44
A


ATOM
3454
O
VAL
A
323
−56.333
1.144
−17.7
1
32.87
A


ATOM
3455
N
VAL
A
324
−57.626
−0.146
−16.375
1
34.69
A


ATOM
3456
CA
VAL
A
324
−57.281
0.558
−15.147
1
33.85
A


ATOM
3457
CB
VAL
A
324
−58.528
1.102
−14.421
1
32.65
A


ATOM
3458
CG1
VAL
A
324
−58.13
1.74
−13.111
1
32.02
A


ATOM
3459
CG2
VAL
A
324
−59.208
2.133
−15.279
1
33.74
A


ATOM
3460
C
VAL
A
324
−56.554
−0.406
−14.223
1
34.46
A


ATOM
3461
O
VAL
A
324
−57.039
−1.506
−13.93
1
34.21
A


ATOM
3462
N
LEU
A
325
−55.375
0.007
−13.78
1
34.48
A


ATOM
3463
CA
LEU
A
325
−54.578
−0.813
−12.891
1
34.12
A


ATOM
3464
CB
LEU
A
325
−53.423
−1.469
−13.66
1
34.48
A


ATOM
3465
CG
LEU
A
325
−52.495
−0.601
−14.513
1
36.04
A


ATOM
3466
CD1
LEU
A
325
−51.692
0.345
−13.641
1
35.91
A


ATOM
3467
CD2
LEU
A
325
−51.554
−1.508
−15.285
1
38.71
A


ATOM
3468
C
LEU
A
325
−54.051
0.011
−11.729
1
33.25
A


ATOM
3469
O
LEU
A
325
−53.828
1.217
−11.848
1
28.42
A


ATOM
3470
N
GLU
A
326
−53.874
−0.66
−10.598
1
34.45
A


ATOM
3471
CA
GLU
A
326
−53.377
−0.029
−9.388
1
34.48
A


ATOM
3472
CB
GLU
A
326
−54.304
−0.347
−8.222
1
36.25
A


ATOM
3473
CG
GLU
A
326
−54.743
0.859
−7.423
1
40.98
A


ATOM
3474
CD
GLU
A
326
−55.57
0.461
−6.218
1
44.97
A


ATOM
3475
OE1
GLU
A
326
−56.491
−0.374
−6.38
1
45.89
A


ATOM
3476
OE2
GLU
A
326
−55.303
0.984
−5.115
1
46.67
A


ATOM
3477
C
GLU
A
326
−51.991
−0.579
−9.099
1
33.25
A


ATOM
3478
O
GLU
A
326
−51.768
−1.792
−9.141
1
32.92
A


ATOM
3479
N
LEU
A
327
−51.058
0.313
−8.805
1
31.12
A


ATOM
3480
CA
LEU
A
327
−49.696
−0.101
−8.514
1
31.14
A


ATOM
3481
CB
LEU
A
327
−48.723
0.566
−9.483
1
29.38
A


ATOM
3482
CG
LEU
A
327
−48.791
0.226
−10.962
1
29.29
A


ATOM
3483
CD1
LEU
A
327
−47.794
1.088
−11.725
1
27.59
A


ATOM
3484
CD2
LEU
A
327
−48.494
−1.25
−11.147
1
27.01
A


ATOM
3485
C
LEU
A
327
−49.285
0.285
−7.109
1
32.57
A


ATOM
3486
O
LEU
A
327
−49.769
1.283
−6.566
1
32.67
A


ATOM
3487
N
GLN
A
328
−48.4
−0.515
−6.517
1
32.21
A


ATOM
3488
CA
GLN
A
328
−47.864
−0.195
−5.201
1
33.66
A


ATOM
3489
CB
GLN
A
328
−48.014
−1.352
−4.206
1
38.45
A


ATOM
3490
CG
GLN
A
328
−47.182
−1.125
−2.922
1
47.56
A


ATOM
3491
CD
GLN
A
328
−47.265
−2.265
−1.904
1
53.01
A


ATOM
3492
OE1
GLN
A
328
−48.321
−2.505
−1.299
1
54.64
A


ATOM
3493
NE2
GLN
A
328
−46.144
−2.968
−1.705
1
50.24
A


ATOM
3494
C
GLN
A
328
−46.381
0.084
−5.418
1
30.77
A


ATOM
3495
O
GLN
A
328
−45.664
−0.758
−5.97
1
31.78
A


ATOM
3496
N
TYR
A
329
−45.924
1.266
−5.01
1
27.23
A


ATOM
3497
CA
TYR
A
329
−44.516
1.607
−5.163
1
24.05
A


ATOM
3498
CB
TYR
A
329
−44.339
3.08
−5.545
1
23.61
A


ATOM
3499
CG
TYR
A
329
−42.899
3.428
−5.839
1
22.91
A


ATOM
3500
CD1
TYR
A
329
−42.14
2.637
−6.702
1
24.27
A


ATOM
3501
CE1
TYR
A
329
−40.792
2.904
−6.929
1
25.43
A


ATOM
3502
CD2
TYR
A
329
−42.278
4.505
−5.214
1
21.99
A


ATOM
3503
CE2
TYR
A
329
−40.926
4.784
−5.431
1
23.92
A


ATOM
3504
CZ
TYR
A
329
−40.188
3.975
−6.285
1
26.19
A


ATOM
3505
OH
TYR
A
329
−38.842
4.197
−6.455
1
25.17
A


ATOM
3506
C
TYR
A
329
−43.782
1.307
−3.863
1
21.92
A


ATOM
3507
O
TYR
A
329
−44.272
1.634
−2.781
1
17.52
A


ATOM
3508
N
THR
A
330
−42.602
0.696
−3.988
1
21.05
A


ATOM
3509
CA
THR
A
330
−41.788
0.304
−2.836
1
19.46
A


ATOM
3510
CB
THR
A
330
−41.143
−1.082
−3.053
1
17.68
A


ATOM
3511
OG1
THR
A
330
−40.181
−0.99
−4.107
1
17.7
A


ATOM
3512
CG2
THR
A
330
−42.19
−2.129
−3.424
1
14.63
A


ATOM
3513
C
THR
A
330
−40.65
1.247
−2.449
1
21.28
A


ATOM
3514
O
THR
A
330
−39.937
0.981
−1.483
1
26.52
A


ATOM
3515
N
GLY
A
331
−40.464
2.341
−3.175
1
20.45
A


ATOM
3516
CA
GLY
A
331
−39.366
3.23
−2.836
1
19.42
A


ATOM
3517
C
GLY
A
331
−39.674
4.419
−1.95
1
19.59
A


ATOM
3518
O
GLY
A
331
−40.833
4.74
−1.692
1
20.07
A


ATOM
3519
N
THR
A
332
−38.615
5.079
−1.488
1
21.73
A


ATOM
3520
CA
THR
A
332
−38.734
6.265
−0.638
1
22.76
A


ATOM
3521
CB
THR
A
332
−38.004
6.068
0.741
1
25.09
A


ATOM
3522
OG1
THR
A
332
−36.59
5.902
0.537
1
20.9
A


ATOM
3523
CG2
THR
A
332
−38.55
4.843
1.462
1
23.58
A


ATOM
3524
C
THR
A
332
−38.098
7.453
−1.359
1
19.89
A


ATOM
3525
O
THR
A
332
−37.855
8.497
−0.766
1
22.26
A


ATOM
3526
N
ASP
A
333
−37.835
7.288
−2.646
1
20.94
A


ATOM
3527
CA
ASP
A
333
−37.186
8.333
−3.426
1
24.06
A


ATOM
3528
CB
ASP
A
333
−36.22
7.695
−4.433
1
21.03
A


ATOM
3529
CG
ASP
A
333
−36.907
6.705
−5.368
1
24.26
A


ATOM
3530
OD1
ASP
A
333
−37.961
6.145
−4.995
1
24.81
A


ATOM
3531
OD2
ASP
A
333
−36.382
6.469
−6.478
1
21.88
A


ATOM
3532
C
ASP
A
333
−38.131
9.295
−4.132
1
27.03
A


ATOM
3533
O
ASP
A
333
−37.694
10.157
−4.897
1
29.68
A


ATOM
3534
N
GLY
A
334
−39.426
9.157
−3.87
1
27.13
A


ATOM
3535
CA
GLY
A
334
−40.389
10.044
−4.492
1
25.95
A


ATOM
3536
C
GLY
A
334
−40.328
11.411
−3.844
1
24.5
A


ATOM
3537
O
GLY
A
334
−39.581
11.607
−2.891
1
25.17
A


ATOM
3538
N
PRO
A
335
−41.086
12.388
−4.35
1
26.13
A


ATOM
3539
CD
PRO
A
335
−41.244
13.713
−3.725
1
26.69
A


ATOM
3540
CA
PRO
A
335
−41.985
12.244
−5.501
1
26.01
A


ATOM
3541
CB
PRO
A
335
−42.662
13.614
−5.586
1
23.99
A


ATOM
3542
CG
PRO
A
335
−42.638
14.105
−4.171
1
25.33
A


ATOM
3543
C
PRO
A
335
−41.182
11.933
−6.759
1
24.96
A


ATOM
3544
O
PRO
A
335
−40.204
12.617
−7.047
1
25.01
A


ATOM
3545
N
CYS
A
336
−41.588
10.903
−7.496
1
24.38
A


ATOM
3546
CA
CYS
A
336
−40.894
10.525
−8.728
1
25.58
A


ATOM
3547
C
CYS
A
336
−41.875
10.098
−9.818
1
26.25
A


ATOM
3548
O
CYS
A
336
−43.023
9.721
−9.54
1
26.06
A


ATOM
3549
CB
CYS
A
336
−39.895
9.389
−8.464
1
22.02
A


ATOM
3550
SG
CYS
A
336
−40.646
7.981
−7.599
1
25.18
A


ATOM
3551
N
LYS
A
337
−41.412
10.157
−11.061
1
25.38
A


ATOM
3552
CA
LYS
A
337
−42.242
9.789
−12.201
1
27.71
A


ATOM
3553
CB
LYS
A
337
−41.713
10.464
−13.483
1
29.76
A


ATOM
3554
CG
LYS
A
337
−42.609
10.297
−14.699
1
29.43
A


ATOM
3555
CD
LYS
A
337
−42.066
11.04
−15.899
1
33.81
A


ATOM
3556
CE
LYS
A
337
−43.023
10.958
−17.089
1
35.15
A


ATOM
3557
NZ
LYS
A
337
−43.267
9.565
−17.559
1
37.12
A


ATOM
3558
C
LYS
A
337
−42.267
8.278
−12.386
1
26.22
A


ATOM
3559
O
LYS
A
337
−41.23
7.616
−12.326
1
26.98
A


ATOM
3560
N
VAL
A
338
−43.455
7.736
−12.615
1
24.74
A


ATOM
3561
CA
VAL
A
338
−43.6
6.305
−12.811
1
25.92
A


ATOM
3562
CB
VAL
A
338
−45.054
5.853
−12.61
1
23.67
A


ATOM
3563
CG1
VAL
A
338
−45.15
4.35
−12.809
1
21.73
A


ATOM
3564
CG2
VAL
A
338
−45.538
6.245
−11.224
1
22.91
A


ATOM
3565
C
VAL
A
338
−43.175
5.883
−14.211
1
28.97
A


ATOM
3566
O
VAL
A
338
−43.677
6.402
−15.207
1
31.92
A


ATOM
3567
N
PRO
A
339
−42.239
4.934
−14.308
1
29.43
A


ATOM
3568
CD
PRO
A
339
−41.476
4.266
−13.247
1
28.71
A


ATOM
3569
CA
PRO
A
339
−41.798
4.485
−15.629
1
28.09
A


ATOM
3570
CB
PRO
A
339
−40.401
3.914
−15.37
1
29.28
A


ATOM
3571
CG
PRO
A
339
−40.132
4.154
−13.877
1
30.81
A


ATOM
3572
C
PRO
A
339
−42.765
3.405
−16.099
1
28.41
A


ATOM
3573
O
PRO
A
339
−42.564
2.224
−15.851
1
31.68
A


ATOM
3574
N
ILE
A
340
−43.834
3.802
−16.759
1
31.06
A


ATOM
3575
CA
ILE
A
340
−44.787
2.814
−17.232
1
29.83
A


ATOM
3576
CB
ILE
A
340
−46.109
2.909
−16.451
1
29.64
A


ATOM
3577
CG2
ILE
A
340
−46.702
4.303
−16.599
1
25.21
A


ATOM
3578
CG1
ILE
A
340
−47.076
1.834
−16.942
1
29.06
A


ATOM
3579
CD1
ILE
A
340
−48.242
1.61
−16.014
1
31.37
A


ATOM
3580
C
ILE
A
340
−45.047
3.012
−18.712
1
29.28
A


ATOM
3581
O
ILE
A
340
−44.963
4.124
−19.224
1
28.22
A


ATOM
3582
N
SER
A
341
−45.348
1.928
−19.408
1
31.18
A


ATOM
3583
CA
SER
A
341
−45.613
2.037
−20.831
1
33.15
A


ATOM
3584
CB
SER
A
341
−44.308
2.294
−21.59
1
31.89
A


ATOM
3585
OG
SER
A
341
−43.348
1.306
−21.28
1
35.88
A


ATOM
3586
C
SER
A
341
−46.302
0.815
−21.397
1
31.64
A


ATOM
3587
O
SER
A
341
−46.187
−0.287
−20.862
1
32.31
A


ATOM
3588
N
SER
A
342
−47.044
1.035
−22.475
1
33.03
A


ATOM
3589
CA
SER
A
342
−47.753
−0.032
−23.161
1
33.02
A


ATOM
3590
CB
SER
A
342
−49.163
0.42
−23.541
1
31.92
A


ATOM
3591
OG
SER
A
342
−49.852
−0.621
−24.209
1
35.86
A


ATOM
3592
C
SER
A
342
−46.949
−0.326
−24.415
1
30.7
A


ATOM
3593
O
SER
A
342
−46.897
0.495
−25.327
1
30.77
A


ATOM
3594
N
VAL
A
343
−46.303
−1.484
−24.443
1
29.8
A


ATOM
3595
CA
VAL
A
343
−45.489
−1.874
−25.586
1
30.71
A


ATOM
3596
CB
VAL
A
343
−44.032
−2.174
−25.171
1
30.39
A


ATOM
3597
CG1
VAL
A
343
−43.351
−0.896
−24.721
1
29.77
A


ATOM
3598
CG2
VAL
A
343
−44.011
−3.207
−24.049
1
27.81
A


ATOM
3599
C
VAL
A
343
−46.069
−3.113
−26.242
1
32.98
A


ATOM
3600
O
VAL
A
343
−46.802
−3.876
−25.604
1
31.51
A


ATOM
3601
N
ALA
A
344
−45.728
−3.302
−27.516
1
34.7
A


ATOM
3602
CA
ALA
A
344
−46.205
−4.433
−28.309
1
34.73
A


ATOM
3603
CB
ALA
A
344
−45.917
−4.185
−29.78
1
35.9
A


ATOM
3604
C
ALA
A
344
−45.577
−5.75
−27.874
1
35.91
A


ATOM
3605
O
ALA
A
344
−46.278
−6.74
−27.647
1
36.55
A


ATOM
3606
N
SER
A
345
−44.253
−5.766
−27.773
1
36.97
A


ATOM
3607
CA
SER
A
345
−43.546
−6.968
−27.353
1
39.5
A


ATOM
3608
CB
SER
A
345
−42.877
−7.666
−28.542
1
39.64
A


ATOM
3609
OG
SER
A
345
−41.551
−7.196
−28.723
1
38.31
A


ATOM
3610
C
SER
A
345
−42.474
−6.586
−26.355
1
40.17
A


ATOM
3611
O
SER
A
345
−42.085
−5.42
−26.265
1
38.89
A


ATOM
3612
N
LEU
A
346
−41.996
−7.579
−25.612
1
41.76
A


ATOM
3613
CA
LEU
A
346
−40.953
−7.354
−24.626
1
41.78
A


ATOM
3614
CB
LEU
A
346
−40.908
−8.521
−23.635
1
39.71
A


ATOM
3615
CG
LEU
A
346
−42.213
−8.708
−22.846
1
38.48
A


ATOM
3616
CD1
LEU
A
346
−42.133
−9.934
−21.952
1
34.83
A


ATOM
3617
CD2
LEU
A
346
−42.476
−7.456
−22.013
1
39.33
A


ATOM
3618
C
LEU
A
346
−39.627
−7.207
−25.367
1
43.43
A


ATOM
3619
O
LEU
A
346
−38.633
−6.752
−24.797
1
43.91
A


ATOM
3620
N
ASN
A
347
−39.632
−7.578
−26.648
1
44.47
A


ATOM
3621
CA
ASN
A
347
−38.441
−7.48
−27.49
1
45.37
A


ATOM
3622
CB
ASN
A
347
−38.566
−8.39
−28.712
1
47.28
A


ATOM
3623
CG
ASN
A
347
−38.732
−9.85
−28.335
1
49.37
A


ATOM
3624
OD1
ASN
A
347
−37.948
−10.39
−27.547
1
49.47
A


ATOM
3625
ND2
ASN
A
347
−39.752
−10.5
−28.9
1
46.9
A


ATOM
3626
C
ASN
A
347
−38.234
−6.04
−27.942
1
44.98
A


ATOM
3627
O
ASN
A
347
−37.175
−5.685
−28.455
1
43.46
A


ATOM
3628
N
ASP
A
348
−39.26
−5.218
−27.755
1
45.11
A


ATOM
3629
CA
ASP
A
348
−39.178
−3.808
−28.106
1
45.32
A


ATOM
3630
CB
ASP
A
348
−39.664
−3.56
−29.535
1
47.04
A


ATOM
3631
CG
ASP
A
348
−39.317
−2.162
−30.031
1
48.74
A


ATOM
3632
OD1
ASP
A
348
−39.557
−1.187
−29.29
1
48.07
A


ATOM
3633
OD2
ASP
A
348
−38.805
−2.036
−31.163
1
50
A


ATOM
3634
C
ASP
A
348
−40.033
−3.011
−27.131
1
43.92
A


ATOM
3635
O
ASP
A
348
−41.255
−3.157
−27.097
1
44.23
A


ATOM
3636
N
LEU
A
349
−39.382
−2.17
−26.337
1
42.36
A


ATOM
3637
CA
LEU
A
349
−40.081
−1.358
−25.351
1
41.12
A


ATOM
3638
CB
LEU
A
349
−39.19
−1.157
−24.125
1
40.88
A


ATOM
3639
CG
LEU
A
349
−39.106
−2.307
−23.114
1
39.94
A


ATOM
3640
CD1
LEU
A
349
−39.349
−3.64
−23.787
1
39.93
A


ATOM
3641
CD2
LEU
A
349
−37.749
−2.271
−22.439
1
37.85
A


ATOM
3642
C
LEU
A
349
−40.541
−0.007
−25.888
1
40.26
A


ATOM
3643
O
LEU
A
349
−40.815
0.911
−25.114
1
42.06
A


ATOM
3644
N
THR
A
350
−40.62
0.118
−27.21
1
37.42
A


ATOM
3645
CA
THR
A
350
−41.077
1.358
−27.817
1
34.7
A


ATOM
3646
CB
THR
A
350
−40.907
1.344
−29.345
1
35.8
A


ATOM
3647
OG1
THR
A
350
−39.513
1.307
−29.662
1
36.6
A


ATOM
3648
CG2
THR
A
350
−41.539
2.588
−29.976
1
33.32
A


ATOM
3649
C
THR
A
350
−42.557
1.505
−27.485
1
34.44
A


ATOM
3650
O
THR
A
350
−43.393
0.69
−27.892
1
34.12
A


ATOM
3651
N
PRO
A
351
−42.898
2.551
−26.73
1
31.71
A


ATOM
3652
CD
PRO
A
351
−42.005
3.605
−26.226
1
28.37
A


ATOM
3653
CA
PRO
A
351
−44.284
2.8
−26.339
1
32.32
A


ATOM
3654
CB
PRO
A
351
−44.195
4.133
−25.606
1
33.14
A


ATOM
3655
CG
PRO
A
351
−42.786
4.129
−25.064
1
32.98
A


ATOM
3656
C
PRO
A
351
−45.195
2.87
−27.555
1
31.96
A


ATOM
3657
O
PRO
A
351
−45.024
3.732
−28.41
1
33.24
A


ATOM
3658
N
VAL
A
352
−46.156
1.957
−27.63
1
33.11
A


ATOM
3659
CA
VAL
A
352
−47.096
1.923
−28.744
1
34.44
A


ATOM
3660
CB
VAL
A
352
−47.109
0.551
−29.416
1
33.1
A


ATOM
3661
CG1
VAL
A
352
−45.734
0.244
−29.97
1
35.53
A


ATOM
3662
CG2
VAL
A
352
−47.539
−0.514
−28.418
1
31.6
A


ATOM
3663
C
VAL
A
352
−48.512
2.254
−28.28
1
38.04
A


ATOM
3664
O
VAL
A
352
−49.339
2.726
−29.068
1
38.22
A


ATOM
3665
N
GLY
A
353
−48.783
1.992
−27.001
1
38.58
A


ATOM
3666
CA
GLY
A
353
−50.085
2.287
−26.435
1
37.9
A


ATOM
3667
C
GLY
A
353
−49.988
3.677
−25.847
1
39.58
A


ATOM
3668
O
GLY
A
353
−48.937
4.31
−25.951
1
39.86
A


ATOM
3669
N
ARG
A
354
−51.055
4.168
−25.228
1
40.79
A


ATOM
3670
CA
ARG
A
354
−50.993
5.507
−24.654
1
41.01
A


ATOM
3671
CB
ARG
A
354
−51.695
6.527
−25.563
1
42.06
A


ATOM
3672
CG
ARG
A
354
−53.174
6.744
−25.258
1
42.39
A


ATOM
3673
CD
ARG
A
354
−53.632
8.126
−25.718
1
44.11
A


ATOM
3674
NE
ARG
A
354
−54.889
8.54
−25.093
1
45.39
A


ATOM
3675
CZ
ARG
A
354
−56.06
7.939
−25.284
1
44.49
A


ATOM
3676
NH1
ARG
A
354
−56.139
6.892
−26.091
1
42.28
A


ATOM
3677
NH2
ARG
A
354
−57.149
8.38
−24.663
1
42.53
A


ATOM
3678
C
ARG
A
354
−51.589
5.594
−23.261
1
39.67
A


ATOM
3679
O
ARG
A
354
−52.609
4.971
−22.957
1
41.21
A


ATOM
3680
N
LEU
A
355
−50.94
6.375
−22.412
1
37.85
A


ATOM
3681
CA
LEU
A
355
−51.418
6.563
−21.059
1
37.76
A


ATOM
3682
CB
LEU
A
355
−50.31
7.14
−20.182
1
38.27
A


ATOM
3683
CG
LEU
A
355
−49.547
6.159
−19.3
1
39.1
A


ATOM
3684
CD1
LEU
A
355
−48.999
5.004
−20.119
1
38.6
A


ATOM
3685
CD2
LEU
A
355
−48.434
6.92
−18.613
1
41.05
A


ATOM
3686
C
LEU
A
355
−52.596
7.517
−21.064
1
36.81
A


ATOM
3687
O
LEU
A
355
−52.565
8.556
−21.726
1
36.66
A


ATOM
3688
N
VAL
A
356
−53.644
7.158
−20.334
1
35.43
A


ATOM
3689
CA
VAL
A
356
−54.808
8.022
−20.241
1
34.17
A


ATOM
3690
CB
VAL
A
356
−56.114
7.218
−20.067
1
33.95
A


ATOM
3691
CG1
VAL
A
356
−57.289
8.162
−19.919
1
31.08
A


ATOM
3692
CG2
VAL
A
356
−56.326
6.324
−21.265
1
30.39
A


ATOM
3693
C
VAL
A
356
−54.586
8.897
−19.021
1
34.44
A


ATOM
3694
O
VAL
A
356
−54.969
10.062
−19.004
1
33.61
A


ATOM
3695
N
THR
A
357
−53.954
8.327
−17.998
1
37.29
A


ATOM
3696
CA
THR
A
357
−53.673
9.073
−16.782
1
38.92
A


ATOM
3697
CB
THR
A
357
−53.289
8.154
−15.62
1
38.58
A


ATOM
3698
OG1
THR
A
357
−54.405
7.327
−15.274
1
41.65
A


ATOM
3699
CG2
THR
A
357
−52.885
8.979
−14.423
1
38.85
A


ATOM
3700
C
THR
A
357
−52.51
10.005
−17.065
1
40.94
A


ATOM
3701
O
THR
A
357
−51.476
9.582
−17.586
1
41.19
A


ATOM
3702
N
VAL
A
358
−52.683
11.274
−16.717
1
42.51
A


ATOM
3703
CA
VAL
A
358
−51.656
12.275
−16.951
1
44.51
A


ATOM
3704
CB
VAL
A
358
−52.221
13.695
−16.749
1
45.68
A


ATOM
3705
CG1
VAL
A
358
−51.084
14.698
−16.685
1
44.38
A


ATOM
3706
CG2
VAL
A
358
−53.157
14.044
−17.888
1
44.87
A


ATOM
3707
C
VAL
A
358
−50.42
12.116
−16.083
1
45.45
A


ATOM
3708
O
VAL
A
358
−50.516
12.061
−14.857
1
44.9
A


ATOM
3709
N
ASN
A
359
−49.27
12.037
−16.75
1
46.22
A


ATOM
3710
CA
ASN
A
359
−47.964
11.908
−16.111
1
44.58
A


ATOM
3711
CB
ASN
A
359
−47.361
13.3
−15.901
1
47.65
A


ATOM
3712
CG
ASN
A
359
−45.855
13.278
−15.824
1
49.05
A


ATOM
3713
OD1
ASN
A
359
−45.249
12.245
−15.534
1
51.83
A


ATOM
3714
ND2
ASN
A
359
−45.236
14.423
−16.091
1
49.2
A


ATOM
3715
C
ASN
A
359
−48.017
11.183
−14.767
1
40.66
A


ATOM
3716
O
ASN
A
359
−47.926
11.8
−13.711
1
40.16
A


ATOM
3717
N
PRO
A
360
−48.182
9.863
−14.791
1
38.08
A


ATOM
3718
CD
PRO
A
360
−48.403
9.047
−15.996
1
38.73
A


ATOM
3719
CA
PRO
A
360
−48.246
9.047
−13.57
1
37
A


ATOM
3720
CB
PRO
A
360
−48.152
7.628
−14.116
1
38.4
A


ATOM
3721
CG
PRO
A
360
−48.919
7.738
−15.401
1
40.41
A


ATOM
3722
C
PRO
A
360
−47.1
9.38
−12.611
1
34.63
A


ATOM
3723
O
PRO
A
360
−45.943
9.41
−13.032
1
33.21
A


ATOM
3724
N
PHE
A
361
−47.425
9.615
−11.338
1
30.6
A


ATOM
3725
CA
PHE
A
361
−46.427
9.97
−10.328
1
27.99
A


ATOM
3726
CB
PHE
A
361
−46.363
11.483
−10.142
1
27.19
A


ATOM
3727
CG
PHE
A
361
−45.181
12.128
−10.78
1
29.31
A


ATOM
3728
CD1
PHE
A
361
−45.116
12.272
−12.161
1
28.76
A


ATOM
3729
CD2
PHE
A
361
−44.138
12.624
−9.996
1
27.22
A


ATOM
3730
CE1
PHE
A
361
−44.03
12.905
−12.761
1
28.47
A


ATOM
3731
CE2
PHE
A
361
−43.053
13.255
−10.58
1
28.43
A


ATOM
3732
CZ
PHE
A
361
−42.998
13.399
−11.975
1
30.74
A


ATOM
3733
C
PHE
A
361
−46.675
9.41
−8.945
1
26.82
A


ATOM
3734
O
PHE
A
361
−47.815
9.177
−8.554
1
29.07
A


ATOM
3735
N
VAL
A
362
−45.59
9.219
−8.204
1
25.53
A


ATOM
3736
CA
VAL
A
362
−45.68
8.802
−6.819
1
23.92
A


ATOM
3737
CB
VAL
A
362
−44.505
7.917
−6.396
1
23.48
A


ATOM
3738
CG1
VAL
A
362
−44.552
7.687
−4.908
1
20.87
A


ATOM
3739
CG2
VAL
A
362
−44.561
6.593
−7.122
1
22.04
A


ATOM
3740
C
VAL
A
362
−45.506
10.173
−6.181
1
27.55
A


ATOM
3741
O
VAL
A
362
−44.428
10.762
−6.276
1
30.15
A


ATOM
3742
N
SER
A
363
−46.556
10.692
−5.553
1
28.86
A


ATOM
3743
CA
SER
A
363
−46.483
12.016
−4.944
1
29.59
A


ATOM
3744
CB
SER
A
363
−47.87
12.636
−4.828
1
28.72
A


ATOM
3745
OG
SER
A
363
−48.283
13.163
−6.077
1
36.53
A


ATOM
3746
C
SER
A
363
−45.817
12.11
−3.589
1
28.95
A


ATOM
3747
O
SER
A
363
−45.652
13.21
−3.055
1
29.26
A


ATOM
3748
N
VAL
A
364
−45.419
10.976
−3.032
1
26.75
A


ATOM
3749
CA
VAL
A
364
−44.802
10.989
−1.716
1
25.13
A


ATOM
3750
CB
VAL
A
364
−45.66
10.186
−0.704
1
25.5
A


ATOM
3751
CG1
VAL
A
364
−47.02
10.858
−0.53
1
19.64
A


ATOM
3752
CG2
VAL
A
364
−45.827
8.743
−1.194
1
21.22
A


ATOM
3753
C
VAL
A
364
−43.386
10.441
−1.69
1
23.92
A


ATOM
3754
O
VAL
A
364
−42.919
9.839
−2.651
1
22.44
A


ATOM
3755
N
ALA
A
365
−42.711
10.672
−0.57
1
23.86
A


ATOM
3756
CA
ALA
A
365
−41.354
10.196
−0.356
1
23.99
A


ATOM
3757
CB
ALA
A
365
−40.453
11.342
0.073
1
22.69
A


ATOM
3758
C
ALA
A
365
−41.419
9.122
0.734
1
24.52
A


ATOM
3759
O
ALA
A
365
−40.444
8.866
1.443
1
26.51
A


ATOM
3760
N
THR
A
366
−42.596
8.517
0.869
1
22.35
A


ATOM
3761
CA
THR
A
366
−42.816
7.445
1.832
1
22.12
A


ATOM
3762
CB
THR
A
366
−43.972
7.768
2.807
1
19.47
A


ATOM
3763
OG1
THR
A
366
−45.131
8.168
2.064
1
20.17
A


ATOM
3764
CG2
THR
A
366
−43.563
8.888
3.758
1
16.23
A


ATOM
3765
C
THR
A
366
−43.171
6.225
1.003
1
21.03
A


ATOM
3766
O
THR
A
366
−43.815
6.345
−0.034
1
23.24
A


ATOM
3767
N
ALA
A
367
−42.747
5.054
1.453
1
21.27
A


ATOM
3768
CA
ALA
A
367
−42.997
3.819
0.723
1
18.86
A


ATOM
3769
CB
ALA
A
367
−42.026
2.753
1.195
1
16.21
A


ATOM
3770
C
ALA
A
367
−44.432
3.309
0.836
1
19.31
A


ATOM
3771
O
ALA
A
367
−45.23
3.793
1.642
1
20.14
A


ATOM
3772
N
ASN
A
368
−44.75
2.323
0.009
1
19.42
A


ATOM
3773
CA
ASN
A
368
−46.074
1.717
−0.01
1
20.52
A


ATOM
3774
CB
ASN
A
368
−46.516
1.354
1.408
1
19.96
A


ATOM
3775
CG
ASN
A
368
−47.391
0.121
1.449
1
21.32
A


ATOM
3776
OD1
ASN
A
368
−48.13
−0.1
2.408
1
23.07
A


ATOM
3777
ND2
ASN
A
368
−47.296
−0.704
0.42
1
23.7
A


ATOM
3778
C
ASN
A
368
−47.1
2.656
−0.633
1
21.13
A


ATOM
3779
O
ASN
A
368
−48.298
2.507
−0.407
1
20.51
A


ATOM
3780
N
ALA
A
369
−46.633
3.637
−1.397
1
20.86
A


ATOM
3781
CA
ALA
A
369
−47.558
4.562
−2.038
1
26
A


ATOM
3782
CB
ALA
A
369
−46.814
5.781
−2.554
1
22.33
A


ATOM
3783
C
ALA
A
369
−48.238
3.824
−3.19
1
29.49
A


ATOM
3784
O
ALA
A
369
−47.588
3.079
−3.929
1
26.52
A


ATOM
3785
N
LYS
A
370
−49.547
4.002
−3.33
1
32.68
A


ATOM
3786
CA
LYS
A
370
−50.252
3.332
−4.413
1
37.07
A


ATOM
3787
CB
LYS
A
370
−51.459
2.542
−3.887
1
41.85
A


ATOM
3788
CG
LYS
A
370
−52.639
3.387
−3.43
1
48.33
A


ATOM
3789
CD
LYS
A
370
−53.853
2.508
−3.138
1
49.69
A


ATOM
3790
CE
LYS
A
370
−55.111
3.341
−2.869
1
51.84
A


ATOM
3791
NZ
LYS
A
370
−56.333
2.481
−2.777
1
50.27
A


ATOM
3792
C
LYS
A
370
−50.692
4.353
−5.452
1
36.03
A


ATOM
3793
O
LYS
A
370
−51.118
5.457
−5.116
1
33.89
A


ATOM
3794
N
VAL
A
371
−50.567
3.973
−6.718
1
36.96
A


ATOM
3795
CA
VAL
A
371
−50.928
4.85
−7.821
1
37.16
A


ATOM
3796
CB
VAL
A
371
−49.671
5.308
−8.586
1
37.16
A


ATOM
3797
CG1
VAL
A
371
−48.714
6.004
−7.637
1
38.71
A


ATOM
3798
CG2
VAL
A
371
−48.985
4.116
−9.211
1
39.91
A


ATOM
3799
C
VAL
A
371
−51.887
4.166
−8.792
1
36.22
A


ATOM
3800
O
VAL
A
371
−51.748
2.981
−9.107
1
35.25
A


ATOM
3801
N
LEU
A
372
−52.857
4.934
−9.266
1
36.24
A


ATOM
3802
CA
LEU
A
372
−53.864
4.44
−10.191
1
35.38
A


ATOM
3803
CB
LEU
A
372
−55.228
4.988
−9.77
1
36.5
A


ATOM
3804
CG
LEU
A
372
−56.509
4.437
−10.394
1
38.49
A


ATOM
3805
CD1
LEU
A
372
−56.58
2.931
−10.204
1
39.04
A


ATOM
3806
CD2
LEU
A
372
−57.707
5.116
−9.733
1
39.15
A


ATOM
3807
C
LEU
A
372
−53.505
4.907
−11.599
1
35.09
A


ATOM
3808
O
LEU
A
372
−53.188
6.076
−11.804
1
34.83
A


ATOM
3809
N
ILE
A
373
−53.559
3.993
−12.564
1
36.15
A


ATOM
3810
CA
ILE
A
373
−53.225
4.31
−13.951
1
34.51
A


ATOM
3811
CB
ILE
A
373
−51.825
3.75
−14.333
1
36.37
A


ATOM
3812
CG2
ILE
A
373
−51.406
4.284
−15.696
1
35.11
A


ATOM
3813
CG1
ILE
A
373
−50.777
4.116
−13.276
1
35.27
A


ATOM
3814
CD1
ILE
A
373
−50.495
5.582
−13.173
1
34.41
A


ATOM
3815
C
ILE
A
373
−54.222
3.73
−14.957
1
34.65
A


ATOM
3816
O
ILE
A
373
−54.636
2.582
−14.849
1
33.98
A


ATOM
3817
N
GLU
A
374
−54.597
4.54
−15.938
1
37.03
A


ATOM
3818
CA
GLU
A
374
−55.502
4.106
−16.996
1
37.04
A


ATOM
3819
CB
GLU
A
374
−56.758
4.998
−17.026
1
37.83
A


ATOM
3820
CG
GLU
A
374
−57.735
4.689
−18.156
1
39.7
A


ATOM
3821
CD
GLU
A
374
−59.08
5.397
−18.006
1
41.38
A


ATOM
3822
OE1
GLU
A
374
−59.111
6.524
−17.47
1
44.84
A


ATOM
3823
OE2
GLU
A
374
−60.11
4.832
−18.437
1
40.46
A


ATOM
3824
C
GLU
A
374
−54.71
4.202
−18.308
1
35.72
A


ATOM
3825
O
GLU
A
374
−54.08
5.222
−18.583
1
31.75
A


ATOM
3826
N
LEU
A
375
−54.719
3.123
−19.089
1
37.32
A


ATOM
3827
CA
LEU
A
375
−53.996
3.072
−20.366
1
38.65
A


ATOM
3828
CB
LEU
A
375
−52.814
2.091
−20.322
1
37.96
A


ATOM
3829
CG
LEU
A
375
−51.592
2.178
−19.427
1
38.6
A


ATOM
3830
CD1
LEU
A
375
−51.971
1.97
−17.971
1
40.96
A


ATOM
3831
CD2
LEU
A
375
−50.624
1.09
−19.87
1
36.52
A


ATOM
3832
C
LEU
A
375
−54.847
2.608
−21.541
1
38.38
A


ATOM
3833
O
LEU
A
375
−55.836
1.88
−21.384
1
37
A


ATOM
3834
N
GLU
A
376
−54.425
3.02
−22.729
1
39.47
A


ATOM
3835
CA
GLU
A
376
−55.077
2.594
−23.958
1
43.74
A


ATOM
3836
CB
GLU
A
376
−55.513
3.786
−24.817
1
46.07
A


ATOM
3837
CG
GLU
A
376
−56.041
3.366
−26.195
1
51.12
A


ATOM
3838
CD
GLU
A
376
−56.707
4.503
−26.941
1
53.35
A


ATOM
3839
OE1
GLU
A
376
−57.76
4.97
−26.459
1
55.13
A


ATOM
3840
OE2
GLU
A
376
−56.184
4.938
−27.994
1
53.92
A


ATOM
3841
C
GLU
A
376
−54.034
1.761
−24.706
1
43.78
A


ATOM
3842
O
GLU
A
376
−53.295
2.281
−25.559
1
43.14
A


ATOM
3843
N
PRO
A
377
−53.945
0.457
−24.374
1
41.78
A


ATOM
3844
CD
PRO
A
377
−54.792
−0.297
−23.43
1
40.55
A


ATOM
3845
CA
PRO
A
377
−52.983
−0.435
−25.022
1
40.61
A


ATOM
3846
CB
PRO
A
377
−53.082
−1.704
−24.189
1
39.5
A


ATOM
3847
CG
PRO
A
377
−54.523
−1.742
−23.833
1
40.14
A


ATOM
3848
C
PRO
A
377
−53.403
−0.673
−26.463
1
40.85
A


ATOM
3849
O
PRO
A
377
−54.575
−0.496
−26.811
1
40.53
A


ATOM
3850
N
PRO
A
378
−52.454
−1.055
−27.33
1
40.26
A


ATOM
3851
CD
PRO
A
378
−51.013
−1.287
−27.127
1
38.43
A


ATOM
3852
CA
PRO
A
378
−52.833
−1.299
−28.723
1
39.63
A


ATOM
3853
CB
PRO
A
378
−51.489
−1.55
−29.399
1
38.71
A


ATOM
3854
CG
PRO
A
378
−50.67
−2.161
−28.302
1
37.56
A


ATOM
3855
C
PRO
A
378
−53.78
−2.5
−28.823
1
39.1
A


ATOM
3856
O
PRO
A
378
−54.044
−3.178
−27.829
1
40.34
A


ATOM
3857
N
PHE
A
379
−54.307
−2.754
−30.012
1
38.56
A


ATOM
3858
CA
PHE
A
379
−55.195
−3.892
−30.196
1
38.41
A


ATOM
3859
CB
PHE
A
379
−56.082
−3.676
−31.42
1
37.4
A


ATOM
3860
CG
PHE
A
379
−57.349
−2.94
−31.117
1
37.23
A


ATOM
3861
CD1
PHE
A
379
−58.406
−3.585
−30.477
1
36.17
A


ATOM
3862
CD2
PHE
A
379
−57.483
−1.593
−31.443
1
36.88
A


ATOM
3863
CE1
PHE
A
379
−59.577
−2.9
−30.166
1
33.86
A


ATOM
3864
CE2
PHE
A
379
−58.653
−0.898
−31.134
1
35.77
A


ATOM
3865
CZ
PHE
A
379
−59.701
−1.557
−30.494
1
33.66
A


ATOM
3866
C
PHE
A
379
−54.339
−5.14
−30.37
1
39.24
A


ATOM
3867
O
PHE
A
379
−53.237
−5.071
−30.914
1
39.2
A


ATOM
3868
N
GLY
A
380
−54.844
−6.279
−29.909
1
38.42
A


ATOM
3869
CA
GLY
A
380
−54.078
−7.501
−30.017
1
37.47
A


ATOM
3870
C
GLY
A
380
−53.242
−7.674
−28.765
1
38.09
A


ATOM
3871
O
GLY
A
380
−53.705
−7.375
−27.666
1
39.96
A


ATOM
3872
N
ASP
A
381
−52.009
−8.141
−28.921
1
37.08
A


ATOM
3873
CA
ASP
A
381
−51.127
−8.354
−27.78
1
36.79
A


ATOM
3874
CB
ASP
A
381
−50.24
−9.573
−28.028
1
38.65
A


ATOM
3875
CG
ASP
A
381
−50.833
−10.85
−27.465
1
42.76
A


ATOM
3876
OD1
ASP
A
381
−52.079
−10.967
−27.421
1
43.71
A


ATOM
3877
OD2
ASP
A
381
−50.048
−11.742
−27.075
1
44.65
A


ATOM
3878
C
ASP
A
381
−50.256
−7.147
−27.448
1
35.55
A


ATOM
3879
O
ASP
A
381
−49.868
−6.378
−28.323
1
36.19
A


ATOM
3880
N
SER
A
382
−49.959
−6.992
−26.166
1
31.97
A


ATOM
3881
CA
SER
A
382
−49.138
−5.898
−25.689
1
29.45
A


ATOM
3882
CB
SER
A
382
−49.901
−4.572
−25.776
1
28.32
A


ATOM
3883
OG
SER
A
382
−50.866
−4.465
−24.739
1
25.09
A


ATOM
3884
C
SER
A
382
−48.784
−6.19
−24.237
1
30.2
A


ATOM
3885
O
SER
A
382
−49.327
−7.109
−23.623
1
28.41
A


ATOM
3886
N
TYR
A
383
−47.873
−5.403
−23.686
1
30.32
A


ATOM
3887
CA
TYR
A
383
−47.47
−5.598
−22.308
1
32.14
A


ATOM
3888
CB
TYR
A
383
−46.064
−6.203
−22.237
1
33.07
A


ATOM
3889
CG
TYR
A
383
−45.92
−7.574
−22.862
1
37.36
A


ATOM
3890
CD1
TYR
A
383
−46.291
−8.731
−22.17
1
38.95
A


ATOM
3891
CE1
TYR
A
383
−46.125
−10.002
−22.744
1
41.13
A


ATOM
3892
CD2
TYR
A
383
−45.389
−7.716
−24.14
1
39.47
A


ATOM
3893
CE2
TYR
A
383
−45.224
−8.977
−24.722
1
44.33
A


ATOM
3894
CZ
TYR
A
383
−45.59
−10.114
−24.023
1
42.11
A


ATOM
3895
OH
TYR
A
383
−45.415
−11.347
−24.619
1
42.76
A


ATOM
3896
C
TYR
A
383
−47.464
−4.262
−21.596
1
31.89
A


ATOM
3897
O
TYR
A
383
−47.199
−3.22
−22.207
1
31.24
A


ATOM
3898
N
ILE
A
384
−47.785
−4.301
−20.307
1
30.93
A


ATOM
3899
CA
ILE
A
384
−47.764
−3.111
−19.476
1
29.23
A


ATOM
3900
CB
ILE
A
384
−48.92
−3.088
−18.465
1
28.41
A


ATOM
3901
CG2
ILE
A
384
−48.857
−1.809
−17.64
1
25.49
A


ATOM
3902
CG1
ILE
A
384
−50.256
−3.181
−19.199
1
27.33
A


ATOM
3903
CD1
ILE
A
384
−51.453
−3.191
−18.273
1
30.87
A


ATOM
3904
C
ILE
A
384
−46.453
−3.271
−18.726
1
29.99
A


ATOM
3905
O
ILE
A
384
−46.301
−4.191
−17.914
1
30.14
A


ATOM
3906
N
VAL
A
385
−45.494
−2.403
−19.022
1
27.27
A


ATOM
3907
CA
VAL
A
385
−44.204
−2.491
−18.374
1
25.77
A


ATOM
3908
CB
VAL
A
385
−43.071
−2.535
−19.409
1
26.78
A


ATOM
3909
CG1
VAL
A
385
−41.71
−2.513
−18.701
1
24.17
A


ATOM
3910
CG2
VAL
A
385
−43.212
−3.787
−20.257
1
25.13
A


ATOM
3911
C
VAL
A
385
−43.96
−1.341
−17.415
1
26.53
A


ATOM
3912
O
VAL
A
385
−44.205
−0.175
−17.731
1
25.21
A


ATOM
3913
N
VAL
A
386
−43.47
−1.692
−16.234
1
23.93
A


ATOM
3914
CA
VAL
A
386
−43.18
−0.71
−15.214
1
22.01
A


ATOM
3915
CB
VAL
A
386
−44.14
−0.867
−14.037
1
22.1
A


ATOM
3916
CG1
VAL
A
386
−43.914
0.242
−13.026
1
14.98
A


ATOM
3917
CG2
VAL
A
386
−45.571
−0.874
−14.56
1
22.7
A


ATOM
3918
C
VAL
A
386
−41.749
−0.907
−14.747
1
20.25
A


ATOM
3919
O
VAL
A
386
−41.335
−2.028
−14.466
1
17.33
A


ATOM
3920
N
GLY
A
387
−41
0.189
−14.69
1
19.86
A


ATOM
3921
CA
GLY
A
387
−39.617
0.131
−14.264
1
24.46
A


ATOM
3922
C
GLY
A
387
−38.686
−0.273
−15.388
1
27.99
A


ATOM
3923
O
GLY
A
387
−39.112
−0.476
−16.526
1
28.8
A


ATOM
3924
N
ARG
A
388
−37.403
−0.385
−15.074
1
30.3
A


ATOM
3925
CA
ARG
A
388
−36.43
−0.78
−16.076
1
33.92
A


ATOM
3926
CB
ARG
A
388
−35.942
0.453
−16.855
1
39.24
A


ATOM
3927
CG
ARG
A
388
−34.92
0.152
−17.972
1
47.47
A


ATOM
3928
CD
ARG
A
388
−35.308
−1.085
−18.811
1
52.19
A


ATOM
3929
NE
ARG
A
388
−34.349
−1.376
−19.881
1
54.83
A


ATOM
3930
CZ
ARG
A
388
−34.369
−2.48
−20.626
1
55.84
A


ATOM
3931
NH1
ARG
A
388
−35.298
−3.401
−20.415
1
57.41
A


ATOM
3932
NH2
ARG
A
388
−33.467
−2.664
−21.586
1
55.45
A


ATOM
3933
C
ARG
A
388
−35.264
−1.495
−15.416
1
32.89
A


ATOM
3934
O
ARG
A
388
−35.234
−1.657
−14.195
1
33.14
A


ATOM
3935
N
GLY
A
389
−34.31
−1.932
−16.231
1
31.69
A


ATOM
3936
CA
GLY
A
389
−33.151
−2.619
−15.706
1
29.14
A


ATOM
3937
C
GLY
A
389
−33.562
−3.881
−14.995
1
27.89
A


ATOM
3938
O
GLY
A
389
−34.457
−4.596
−15.44
1
27.07
A


ATOM
3939
N
GLU
A
390
−32.912
−4.15
−13.875
1
29.38
A


ATOM
3940
CA
GLU
A
390
−33.207
−5.341
−13.1
1
27.7
A


ATOM
3941
CB
GLU
A
390
−32.043
−5.64
−12.162
1
27.62
A


ATOM
3942
CG
GLU
A
390
−30.828
−6.151
−12.9
1
31.45
A


ATOM
3943
CD
GLU
A
390
−29.698
−6.554
−11.985
1
31.31
A


ATOM
3944
OE1
GLU
A
390
−29.968
−7.069
−10.874
1
30.56
A


ATOM
3945
OE2
GLU
A
390
−28.533
−6.373
−12.394
1
31.78
A


ATOM
3946
C
GLU
A
390
−34.511
−5.262
−12.313
1
27.76
A


ATOM
3947
O
GLU
A
390
−34.99
−6.28
−11.816
1
27.45
A


ATOM
3948
N
GLN
A
391
−35.091
−4.069
−12.19
1
26.73
A


ATOM
3949
CA
GLN
A
391
−36.347
−3.952
−11.453
1
28.35
A


ATOM
3950
CB
GLN
A
391
−36.249
−2.897
−10.336
1
26
A


ATOM
3951
CG
GLN
A
391
−35.451
−1.656
−10.647
1
24.34
A


ATOM
3952
CD
GLN
A
391
−33.956
−1.915
−10.696
1
25.55
A


ATOM
3953
OE1
GLN
A
391
−33.421
−2.706
−9.919
1
23.68
A


ATOM
3954
NE2
GLN
A
391
−33.271
−1.23
−11.6
1
27.54
A


ATOM
3955
C
GLN
A
391
−37.553
−3.678
−12.353
1
29.91
A


ATOM
3956
O
GLN
A
391
−38.581
−3.153
−11.917
1
30.87
A


ATOM
3957
N
GLN
A
392
−37.423
−4.055
−13.618
1
28.06
A


ATOM
3958
CA
GLN
A
392
−38.503
−3.873
−14.562
1
24.73
A


ATOM
3959
CB
GLN
A
392
−37.952
−3.802
−15.98
1
25
A


ATOM
3960
CG
GLN
A
392
−39.006
−3.982
−17.057
1
24.39
A


ATOM
3961
CD
GLN
A
392
−38.419
−3.895
−18.448
1
26.15
A


ATOM
3962
OE1
GLN
A
392
−37.832
−2.873
−18.822
1
24.56
A


ATOM
3963
NE2
GLN
A
392
−38.567
−4.968
−19.225
1
24.99
A


ATOM
3964
C
GLN
A
392
−39.442
−5.058
−14.443
1
24.52
A


ATOM
3965
O
GLN
A
392
−38.996
−6.2
−14.347
1
23.24
A


ATOM
3966
N
ILE
A
393
−40.74
−4.78
−14.433
1
23.15
A


ATOM
3967
CA
ILE
A
393
−41.751
−5.828
−14.356
1
23.89
A


ATOM
3968
CB
ILE
A
393
−42.533
−5.792
−13.034
1
20.25
A


ATOM
3969
CG2
ILE
A
393
−41.573
−5.868
−11.862
1
22.08
A


ATOM
3970
CG1
ILE
A
393
−43.366
−4.513
−12.962
1
16.82
A


ATOM
3971
CD1
ILE
A
393
−44.293
−4.463
−11.794
1
13.69
A


ATOM
3972
C
ILE
A
393
−42.736
−5.577
−15.482
1
26.36
A


ATOM
3973
O
ILE
A
393
−42.853
−4.456
−15.977
1
28.56
A


ATOM
3974
N
ASN
A
394
−43.445
−6.616
−15.891
1
29.16
A


ATOM
3975
CA
ASN
A
394
−44.416
−6.453
−16.958
1
33.73
A


ATOM
3976
CB
ASN
A
394
−43.8
−6.821
−18.311
1
33.54
A


ATOM
3977
CG
ASN
A
394
−43.282
−8.24
−18.345
1
37.25
A


ATOM
3978
OD1
ASN
A
394
−44.056
−9.201
−18.284
1
35.83
A


ATOM
3979
ND2
ASN
A
394
−41.96
−8.383
−18.439
1
36.25
A


ATOM
3980
C
ASN
A
394
−45.639
−7.304
−16.705
1
33.07
A


ATOM
3981
O
ASN
A
394
−45.628
−8.202
−15.866
1
33.86
A


ATOM
3982
N
HIS
A
395
−46.704
−6.99
−17.424
1
32.43
A


ATOM
3983
CA
HIS
A
395
−47.938
−7.732
−17.304
1
31.53
A


ATOM
3984
CB
HIS
A
395
−48.936
−7.009
−16.4
1
30.12
A


ATOM
3985
CG
HIS
A
395
−50.17
−7.808
−16.108
1
32.43
A


ATOM
3986
CD2
HIS
A
395
−50.724
−8.196
−14.933
1
31.12
A


ATOM
3987
ND1
HIS
A
395
−50.995
−8.296
−17.101
1
30.44
A


ATOM
3988
CE1
HIS
A
395
−52.003
−8.949
−16.551
1
29.5
A


ATOM
3989
NE2
HIS
A
395
−51.863
−8.903
−15.238
1
34.58
A


ATOM
3990
C
HIS
A
395
−48.471
−7.81
−18.714
1
32.74
A


ATOM
3991
O
HIS
A
395
−48.61
−6.786
−19.389
1
30.82
A


ATOM
3992
N
HIS
A
396
−48.736
−9.034
−19.162
1
34.05
A


ATOM
3993
CA
HIS
A
396
−49.249
−9.258
−20.502
1
30.98
A


ATOM
3994
CB
HIS
A
396
−49.193
−10.751
−20.851
1
30.51
A


ATOM
3995
CG
HIS
A
396
−49.803
−11.08
−22.176
1
29.82
A


ATOM
3996
CD2
HIS
A
396
−49.321
−10.952
−23.435
1
29.9
A


ATOM
3997
ND1
HIS
A
396
−51.098
−11.534
−22.306
1
29.99
A


ATOM
3998
CE1
HIS
A
396
−51.388
−11.67
−23.588
1
29.56
A


ATOM
3999
NE2
HIS
A
396
−50.328
−11.322
−24.294
1
31.1
A


ATOM
4000
C
HIS
A
396
−50.673
−8.739
−20.615
1
28.74
A


ATOM
4001
O
HIS
A
396
−51.353
−8.525
−19.61
1
24.91
A


ATOM
4002
N
TRP
A
397
−51.112
−8.523
−21.846
1
28.3
A


ATOM
4003
CA
TRP
A
397
−52.454
−8.036
−22.089
1
31.79
A


ATOM
4004
CB
TRP
A
397
−52.543
−6.552
−21.725
1
34.26
A


ATOM
4005
CG
TRP
A
397
−53.931
−6.04
−21.737
1
37.71
A


ATOM
4006
CD2
TRP
A
397
−54.898
−6.191
−20.695
1
39.81
A


ATOM
4007
CE2
TRP
A
397
−56.107
−5.634
−21.165
1
41.46
A


ATOM
4008
CE3
TRP
A
397
−54.861
−6.748
−19.408
1
40.17
A


ATOM
4009
CD1
TRP
A
397
−54.567
−5.411
−22.766
1
39.17
A


ATOM
4010
NE1
TRP
A
397
−55.876
−5.164
−22.432
1
41.69
A


ATOM
4011
CZ2
TRP
A
397
−57.275
−5.617
−20.393
1
42.7
A


ATOM
4012
CZ3
TRP
A
397
−56.021
−6.733
−18.639
1
41.83
A


ATOM
4013
CH2
TRP
A
397
−57.212
−6.169
−19.136
1
44.03
A


ATOM
4014
C
TRP
A
397
−52.831
−8.239
−23.551
1
32.49
A


ATOM
4015
O
TRP
A
397
−51.973
−8.22
−24.434
1
32.14
A


ATOM
4016
N
HIS
A
398
−54.117
−8.457
−23.798
1
33.24
A


ATOM
4017
CA
HIS
A
398
−54.608
−8.645
−25.153
1
34.78
A


ATOM
4018
CB
HIS
A
398
−54.824
−10.12
−25.471
1
39.72
A


ATOM
4019
CG
HIS
A
398
−55.384
−10.347
−26.841
1
44.25
A


ATOM
4020
CD2
HIS
A
398
−56.658
−10.527
−27.263
1
45.49
A


ATOM
4021
ND1
HIS
A
398
−54.607
−10.302
−27.979
1
46.12
A


ATOM
4022
CE1
HIS
A
398
−55.379
−10.441
−29.043
1
46.48
A


ATOM
4023
NE2
HIS
A
398
−56.628
−10.578
−28.635
1
46.09
A


ATOM
4024
C
HIS
A
398
−55.927
−7.92
−25.317
1
32.79
A


ATOM
4025
O
HIS
A
398
−56.874
−8.181
−24.584
1
30.08
A


ATOM
4026
N
LYS
A
399
−55.982
−7.012
−26.285
1
33.29
A


ATOM
4027
CA
LYS
A
399
−57.186
−6.238
−26.542
1
33.44
A


ATOM
4028
CB
LYS
A
399
−56.826
−4.766
−26.718
1
32.16
A


ATOM
4029
CG
LYS
A
399
−58.005
−3.838
−26.977
1
31.71
A


ATOM
4030
CD
LYS
A
399
−57.54
−2.384
−27.007
1
32.16
A


ATOM
4031
CE
LYS
A
399
−58.701
−1.416
−27.187
1
34.69
A


ATOM
4032
NZ
LYS
A
399
−58.237
−0.001
−27.321
1
33.07
A


ATOM
4033
C
LYS
A
399
−57.896
−6.74
−27.785
1
35.18
A


ATOM
4034
O
LYS
A
399
−57.387
−6.588
−28.894
1
36.94
A


ATOM
4035
N
LYS
A
400
−59.066
−7.346
−27.592
1
36.95
A


ATOM
4036
CA
LYS
A
400
−59.87
−7.857
−28.698
1
39.25
A


ATOM
4037
CB
LYS
A
400
−61.003
−8.732
−28.148
1
41.37
A


ATOM
4038
CG
LYS
A
400
−61.741
−8.099
−26.965
1
46.59
A


ATOM
4039
CD
LYS
A
400
−62.899
−8.965
−26.468
1
52.09
A


ATOM
4040
CE
LYS
A
400
−63.505
−8.403
−25.174
1
54.62
A


ATOM
4041
NZ
LYS
A
400
−64.016
−7
−25.315
1
56.17
A


ATOM
4042
C
LYS
A
400
−60.462
−6.701
−29.522
1
39.94
A


ATOM
4043
O
LYS
A
400
−60.759
−5.634
−28.93
1
34.44
A


ATOM
4044
OXT
LYS
A
400
−60.638
−6.886
−30.753
1
41.62
A


ATOM
4045
NA
NA

1
−50.542
0.414
8.067
1
29.56


ATOM
4046
OH2
WAT
W
3
−43.941
4.79
−2.357
1
18.34
W


ATOM
4047
OH2
WAT
W
4
−19.646
−17.689
40.827
1
19.49
W


ATOM
4048
OH2
WAT
W
5
1.086
−6.228
36.071
1
34.43
W


ATOM
4049
OH2
WAT
W
6
−9.954
−24.698
28.495
1
10.6
W


ATOM
4050
OH2
WAT
W
7
−31.62
0.121
−5.52
1
5.87
W


ATOM
4051
OH2
WAT
W
8
−10.669
16.435
6.361
1
30.99
W


ATOM
4052
OH2
WAT
W
9
−11.045
−8.113
17.167
1
20.54
W


ATOM
4053
OH2
WAT
W
10
−24.036
−12.754
10.102
1
8.53
W


ATOM
4054
OH2
WAT
W
11
−57.533
−2.732
−11.116
1
38.68
W


ATOM
4055
OH2
WAT
W
12
−46.647
−12.435
−27.215
1
32.63
W


ATOM
4056
OH2
WAT
W
13
−19.137
−11.152
−8.672
1
20.57
W


ATOM
4057
OH2
WAT
W
14
−38.72
0.64
1.016
1
21.59
W


ATOM
4058
OH2
WAT
W
15
−34.334
9.295
−12.941
1
40.13
W


ATOM
4059
OH2
WAT
W
16
−35.475
12.109
15.787
1
17.9
W


ATOM
4060
OH2
WAT
W
17
−27.27
−10.354
−10.034
1
26.77
W


ATOM
4061
OH2
WAT
W
18
−50.153
9.273
−6.596
1
21.61
W


ATOM
4062
OH2
WAT
W
19
−54.458
7.632
−7.685
1
35.4
W


ATOM
4063
OH2
WAT
W
20
−42.711
−10.749
−26.432
1
39.16
W


ATOM
4064
OH2
WAT
W
21
−16.864
−7.007
−6.017
1
21.82
W


ATOM
4065
OH2
WAT
W
22
−7.613
−12.554
28.423
1
22
W


ATOM
4066
OH2
WAT
W
23
−4.625
−8.443
6.447
1
27.94
W


ATOM
4067
OH2
WAT
W
24
−33.146
0.338
−7.652
1
15.92
W


ATOM
4068
OH2
WAT
W
25
−42.132
9.317
14.521
1
36.69
W


ATOM
4069
OH2
WAT
W
26
−10.584
−4.63
6.868
1
34.03
W


ATOM
4070
OH2
WAT
W
27
−40.841
−2.544
−11.395
1
26.86
W


ATOM
4071
OH2
WAT
W
28
4.096
−30.579
13.731
1
29.83
W


ATOM
4072
OH2
WAT
W
29
−9.395
0.621
17.188
1
22.04
W


ATOM
4073
OH2
WAT
W
30
−22.711
−3.107
−14.633
1
15.28
W


ATOM
4074
OH2
WAT
W
31
−20.21
−8.921
−0.257
1
28.57
W


ATOM
4075
OH2
WAT
W
32
−42.448
−9.379
−14.425
1
17.41
W


ATOM
4076
OH2
WAT
W
33
1.681
−10.01
5.93
1
25.24
W


ATOM
4077
OH2
WAT
W
34
−3.312
−26.223
30.059
1
21.14
W


ATOM
4078
OH2
WAT
W
36
−25.451
−0.275
−7.377
1
28.12
W


ATOM
4079
OH2
WAT
W
37
−29.078
0.682
15.655
1
26.86
W


ATOM
4080
OH2
WAT
W
38
−7.919
−26.525
9.384
1
40.51
W


ATOM
4081
OH2
WAT
W
39
−45.092
−5.436
−1.04
1
16.01
W


ATOM
4082
OH2
WAT
W
40
−16.396
−27.635
29.589
1
11.74
W


ATOM
4083
OH2
WAT
W
41
−21.452
−7.414
−13.818
1
20.73
W


ATOM
4084
OH2
WAT
W
42
−14.403
7.628
−9.634
1
25.09
W


ATOM
4085
OH2
WAT
W
43
−23.999
−0.875
−14.736
1
16.91
W


ATOM
4086
OH2
WAT
W
44
−24.561
9.919
18.966
1
35.63
W


ATOM
4087
OH2
WAT
W
45
−16.117
−17.767
19.742
1
18.5
W


ATOM
4088
OH2
WAT
W
46
−11.106
−19.809
12.775
1
25.09
W


ATOM
4089
OH2
WAT
W
47
2.761
−21.103
26.079
1
26.92
W


ATOM
4090
OH2
WAT
W
48
−35.733
−6.866
−4.106
1
12.52
W


ATOM
4091
OH2
WAT
W
50
−49.069
−8.508
11.032
1
24.85
W


ATOM
4092
OH2
WAT
W
51
−11.59
−13.529
14.232
1
29.89
W


ATOM
4093
OH2
WAT
W
52
−61.671
7.809
−18.787
1
42.95
W


ATOM
4094
OH2
WAT
W
53
−41.977
−8.045
10.834
1
11.86
W


ATOM
4095
OH2
WAT
W
54
−15.503
−9.76
26.774
1
28.9
W


ATOM
4096
OH2
WAT
W
55
−5.798
−0.618
4.361
1
22.56
W


ATOM
4097
OH2
WAT
W
56
−25.556
−3.307
19.778
1
27.17
W


ATOM
4098
OH2
WAT
W
58
−1.854
−29.093
8.502
1
15.68
W


ATOM
4099
OH2
WAT
W
59
−43.778
6.888
−18.587
1
31.84
W


ATOM
4100
OH2
WAT
W
60
−12.379
−28.435
41.824
1
32.9
W


ATOM
4101
OH2
WAT
W
61
−35.591
−8.493
−2.033
1
20.68
W


ATOM
4102
OH2
WAT
W
63
−22.684
−3.858
18.067
1
39.8
W


ATOM
4103
OH2
WAT
W
64
−59.578
−7.443
−11.929
1
37.29
W


ATOM
4104
OH2
WAT
W
65
−2.537
−8.06
42.609
1
38.58
W


ATOM
4105
OH2
WAT
W
67
−30.333
8.228
12.443
1
34.69
W


ATOM
4106
OH2
WAT
W
71
−27.769
15.504
−2.627
1
45.02
W


ATOM
4107
OH2
WAT
W
72
−44.297
12.041
8.685
1
40.86
W


ATOM
4108
OH2
WAT
W
73
−13.211
−8.423
34.777
1
31.99
W


ATOM
4109
OH2
WAT
W
75
−19.2
−12.653
36.524
1
36.24
W


ATOM
4110
OH2
WAT
W
79
−27.954
−10.377
3.053
1
18.8
W


ATOM
4111
OH2
WAT
W
80
−34.448
10.284
−8.574
1
21.89
W


ATOM
4112
OH2
WAT
W
81
−33.81
13.106
−1.454
1
38.55
W


ATOM
4113
OH2
WAT
W
82
−16.27
10.2
20.687
1
49.84
W


ATOM
4114
OH2
WAT
W
83
−38.181
0.526
−20.016
1
39.28
W


ATOM
4115
OH2
WAT
W
84
1.364
−36.226
19.122
1
29.53
W


ATOM
4116
OH2
WAT
W
85
−3.268
−38.021
27.054
1
29.55
W


ATOM
4117
OH2
WAT
W
86
−48.693
8.433
−23.867
1
33.43
W


ATOM
4118
OH2
WAT
W
87
−11.887
−2.729
25.847
1
29.25
W


ATOM
4119
OH2
WAT
W
89
−42.718
−14.193
7.358
1
26.37
W


ATOM
4120
OH2
WAT
W
90
14.268
−13.259
37.789
1
27.13
W


ATOM
4121
OH2
WAT
W
91
−51.051
−9.194
−31.598
1
24.62
W


ATOM
4122
OH2
WAT
W
92
6.827
−25.669
30.531
1
45.3
W


ATOM
4123
OH2
WAT
W
93
−7.182
−14.475
9.866
1
29.24
W


ATOM
4124
OH2
WAT
W
94
4.95
−12.092
16.655
1
17.37
W


ATOM
4125
OH2
WAT
W
95
5.197
−8.421
26.118
1
33.31
W


ATOM
4126
OH2
WAT
W
96
−28.026
−17.054
28.443
1
41.37
W


ATOM
4127
OH2
WAT
W
98
−7.529
7.352
1.881
1
36.14
W


ATOM
4128
OH2
WAT
W
99
−47.018
6.863
17.868
1
32.12
W


ATOM
4129
OH2
WAT
W
100
−12.153
3.851
−9.904
1
37.54
W


ATOM
4130
OH2
WAT
W
101
−28.015
−28.318
27.269
1
39.42
W


ATOM
4131
OH2
WAT
W
102
−44.13
−23.562
13.784
1
31.09
W


ATOM
4132
OH2
WAT
W
103
−12.032
−0.104
−8.589
1
31.48
W


ATOM
4133
OH2
WAT
W
104
−3.184
−28.525
37.438
1
34.01
W


ATOM
4134
OH2
WAT
W
105
−45
−18.165
13.362
1
24.92
W


ATOM
4135
OH2
WAT
W
106
3.14
−4.373
20.948
1
31.28
W


ATOM
4136
OH2
WAT
W
107
5.214
−35.144
16.679
1
28.3
W


ATOM
4137
OH2
WAT
W
108
−14.904
−29.092
15.767
1
29.14
W


ATOM
4138
OH2
WAT
W
109
−10.871
7.385
−7.231
1
38.83
W


ATOM
4139
OH2
WAT
W
111
3.837
−13.403
44.67
1
17.58
W


ATOM
4140
OH2
WAT
W
113
1.416
−9.092
13.586
1
34.93
W


ATOM
4141
OH2
WAT
W
115
−34.024
3.785
15.713
1
21.73
W


ATOM
4142
OH2
WAT
W
117
−5.926
13.178
4.491
1
34.28
W


ATOM
4143
OH2
WAT
W
118
−9.712
10.888
−2.079
1
28.67
W


ATOM
4144
OH2
WAT
W
120
−23.559
14.629
−3.288
1
28.54
W


ATOM
4145
OH2
WAT
W
122
−46.401
7.792
9.601
1
29.9
W


ATOM
4146
OH2
WAT
W
123
4.489
−27.789
33.795
1
35.5
W


ATOM
4147
OH2
WAT
W
124
−41.971
−14.555
2.195
1
23.34
W


ATOM
4148
OH2
WAT
W
125
1.529
−26.419
41.363
1
42.16
W


ATOM
4149
OH2
WAT
W
126
−54.535
5.369
−5.331
1
41.52
W


ATOM
4150
OH2
WAT
W
127
−29.834
5.694
10.742
1
22.15
W


ATOM
4151
OH2
WAT
W
131
−25.016
−9.801
3.443
1
27.15
W


ATOM
4152
OH2
WAT
W
132
−45.158
−13.521
2.512
1
34.24
W


ATOM
4153
OH2
WAT
W
134
−53.605
−1.141
−32.217
1
30.09
W


ATOM
4154
OH2
WAT
W
135
−33.135
8.277
14.41
1
33.83
W


ATOM
4155
OH2
WAT
W
141
−27.743
−25.418
15.495
1
34.83
W


ATOM
4156
OH2
WAT
W
142
−47.496
−7.98
7.06
1
28.52
W


ATOM
4157
OH2
WAT
W
143
−41.532
−24.816
17.896
1
44.67
W


ATOM
4158
OH2
WAT
W
144
−30.197
−17.354
9.222
1
25.92
W


ATOM
4159
OH2
WAT
W
146
−18.969
−15.665
13.523
1
44.99
W


ATOM
4160
OH2
WAT
W
147
4.084
−29.484
16.201
1
26.71
W


ATOM
4161
OH2
WAT
W
148
−51.76
−9.627
−11.405
1
25.63
W


ATOM
4162
OH2
WAT
W
149
−9.797
2.005
3.417
1
30
W


ATOM
4163
OH2
WAT
W
150
−50.367
−4.998
−30.382
1
18.32
W


ATOM
4164
OH2
WAT
W
151
−48.037
5.738
3.02
1
31.66
W


ATOM
4165
OH2
WAT
W
152
−32.887
−9.693
−3.002
1
17.39
W


ATOM
4166
OH2
WAT
W
153
−2.606
−7.463
27.853
1
37.76
W


ATOM
4167
OH2
WAT
W
154
4.065
7.213
12.04
1
32.93
W


ATOM
4168
OH2
WAT
W
155
2.121
−14.399
49.961
1
27.98
W


ATOM
4169
OH2
WAT
W
156
−31.712
13.49
−3.435
1
40.65
W


ATOM
4170
OH2
WAT
W
157
−59.365
9.966
−23.221
1
31.27
W


ATOM
4171
OH2
WAT
W
158
−60.818
−3.919
−32.872
1
34.34
W


ATOM
4172
OH2
WAT
W
159
−8.748
−29.386
26.436
1
24.58
W


ATOM
4173
OH2
WAT
W
160
−37.794
9.719
1.924
1
22.76
W


ATOM
4174
OH2
WAT
W
161
−3.91
−39.855
17.062
1
29
W


ATOM
4175
OH2
WAT
W
162
−38.139
11.909
3.82
1
29.1
W


ATOM
4176
OH2
WAT
W
163
−36.45
−6.944
−14.523
1
18.56
W


ATOM
4177
OH2
WAT
W
164
−57.197
11.874
−22.271
1
31.07
W


ATOM
4178
OH2
WAT
W
165
−26.891
−0.921
20.238
1
25.62
W


ATOM
4179
OH2
WAT
W
166
3.165
−5.109
18.495
1
37.61
W


ATOM
4180
OH2
WAT
W
167
−10.745
−12.212
45.915
1
24.8
W


ATOM
4181
OH2
WAT
W
168
−29.697
−18.832
11.574
1
19.28
W


ATOM
4182
OH2
WAT
W
169
−23.691
13.603
20.492
1
19.08
W


ATOM
4183
OH2
WAT
W
170
−3.3
−9.136
31.092
1
22.93
W


ATOM
4184
OH2
WAT
W
171
−20.25
−29.811
32.651
1
33.97
W


ATOM
4185
OH2
WAT
W
173
−19.085
−23.806
48.19
1
36.49
W


ATOM
4186
OH2
WAT
W
174
−40.994
1.49
−18.977
1
32.56
W


ATOM
4187
OH2
WAT
W
176
−17.418
−10.858
40.95
1
40.39
W


ATOM
4188
OH2
WAT
W
177
−21.842
−19.888
37.57
1
26.47
W


ATOM
4189
OH2
WAT
W
178
−22.506
−30.222
16.792
1
30.65
W


ATOM
4190
OH2
WAT
W
179
−19.863
−11.605
6.897
1
39.38
W


ATOM
4191
OH2
WAT
W
180
−13.012
−4.544
9.327
1
33.85
W


ATOM
4192
OH2
WAT
W
181
−25.176
−27.068
38.91
1
37.85
W


ATOM
4193
OH2
WAT
W
183
−65.352
1.208
−16.929
1
29.26
W


ATOM
4194
OH2
WAT
W
184
−19.994
−15.102
−3.378
1
31.19
W


ATOM
4195
OH2
WAT
W
185
−27.082
2.523
18.781
1
43.55
W


ATOM
4196
OH2
WAT
W
186
−5.039
−16.797
48.678
1
35.97
W


ATOM
4197
OH2
WAT
W
187
−12.301
−7.42
32.528
1
37.6
W


ATOM
4198
OH2
WAT
W
188
−16.588
−22.993
42.555
1
34.44
W


ATOM
4199
OH2
WAT
W
189
−39.41
−7.322
−0.594
1
34.16
W


ATOM
4200
OH2
WAT
W
190
−33.056
−6.953
−5.163
1
23.53
W


ATOM
4201
OH2
WAT
W
192
−18.924
−10.295
23.752
1
33.86
W


ATOM
4202
OH2
WAT
W
193
−53.265
−4.866
−26.022
1
27.56
W


ATOM
4203
OH2
WAT
W
194
−10.634
−28.427
10.439
1
21.41
W


ATOM
4204
OH2
WAT
W
195
−26.216
19.288
9.166
1
29.18
W


ATOM
4205
OH2
WAT
W
196
−51.259
0.081
4.271
1
34.01
W


ATOM
4206
OH2
WAT
W
198
−54.493
9.814
−11.966
1
35.64
W


ATOM
4207
OH2
WAT
W
199
−51.515
4.818
11.811
1
32.77
W


ATOM
4208
OH2
WAT
W
201
−19.904
−0.946
−16.34
1
46.53
W


ATOM
4209
OH2
WAT
W
202
−13.94
−4.66
13.551
1
41.06
W


ATOM
4210
OH2
WAT
W
203
−41.991
−9.96
−11.181
1
39.22
W


ATOM
4211
OH2
WAT
W
205
−14.216
−6.746
−6.843
1
53.44
W


ATOM
4212
OH2
WAT
W
206
0.066
−36.97
22.173
1
42.85
W


ATOM
4213
OH2
WAT
W
207
−50.148
10.546
−10.094
1
38.75
W


ATOM
4214
OH2
WAT
W
209
−2.26
−22.521
7.468
1
35.58
W


ATOM
4215
OH2
WAT
W
210
−17.752
−9.352
7.984
1
44.75
W


ATOM
4216
OH2
WAT
W
211
−40.052
19.82
−8.619
1
36.84
W


ATOM
4217
OH2
WAT
W
212
−22.696
14.056
5.646
1
39.6
W


ATOM
4218
OH2
WAT
W
215
−11.847
−4.126
4.173
1
34.71
W


ATOM
4219
OH2
WAT
W
216
−23.564
−14.667
19.777
1
33.88
W


ATOM
4220
OH2
WAT
W
217
−39.052
1.855
−11.372
1
28.38
W


ATOM
4221
OH2
WAT
W
220
−46.305
−5.781
1.333
1
32.23
W


ATOM
4222
OH2
WAT
W
221
−2.553
2.228
8.94
1
32.35
W


ATOM
4223
OH2
WAT
W
225
−12.457
−16.81
11.936
1
47.68
W


ATOM
4224
OH2
WAT
W
226
−33.784
10.504
15.992
1
28.25
W


ATOM
4225
OH2
WAT
W
231
−12.916
12.654
22.606
1
25.49
W


ATOM
4226
OH2
WAT
W
232
−1.305
−3.793
26.802
1
41.27
W


ATOM
4227
OH2
WAT
W
233
0.581
−5.162
29.339
1
36.96
W


ATOM
4228
OH2
WAT
W
234
−40.776
21.479
−0.151
1
45.9
W


ATOM
4229
OH2
WAT
W
235
−20.017
−5.77
−11.939
1
38.79
W


ATOM
4230
OH2
WAT
W
237
11.881
−19.428
41.503
1
33.47
W


ATOM
4231
OH2
WAT
W
238
−54.6
8.984
−5.06
1
37.3
W


ATOM
4232
OH2
WAT
W
241
−52.322
8.463
−9.917
1
35.79
W


ATOM
4233
OH2
WAT
W
242
−4.864
−13.095
5.55
1
30.23
W


ATOM
4234
OH2
WAT
W
243
−43.731
−6.541
−8.789
1
37.23
W


ATOM
4235
OH2
WAT
W
244
−36.38
−10.499
2.089
1
29.07
W


ATOM
4236
OH2
WAT
W
245
1.749
−27.24
15.175
1
29.12
W


ATOM
4237
OH2
WAT
W
246
−27.579
−22.293
36.811
1
29.06
W


ATOM
4238
OH2
WAT
W
247
−59.057
−3.569
−23.512
1
25.71
W


ATOM
4239
OH2
WAT
W
248
−10.112
−28.44
40.611
1
33.55
W


ATOM
4240
OH2
WAT
W
249
−43.565
−12.688
13.94
1
28.84
W


ATOM
4241
OH2
WAT
W
250
−40.113
3.976
19.024
1
31.05
W


ATOM
4242
OH2
WAT
W
251
−12.317
−18.241
41.84
1
24.2
W


ATOM
4243
OH2
WAT
W
252
−47.3
−8.272
14.822
1
42.45
W


ATOM
4244
OH2
WAT
W
253
−46.818
3.216
22.497
1
34.66
W


ATOM
4245
OH2
WAT
W
254
−48.175
−3.685
1.054
1
44.8
W


ATOM
4246
OH2
WAT
W
255
−26.115
11.713
15.208
1
40.98
W


ATOM
4247
OH2
WAT
W
256
0.124
−26.024
31.564
1
20.75
W


ATOM
4248
OH2
WAT
W
257
−4.796
4.473
21.893
1
28.78
W


ATOM
4249
OH2
WAT
W
259
4.803
−14.905
46.635
1
24.03
W


ATOM
4250
OH2
WAT
W
260
−42.149
−20.138
10.772
1
43.71
W


ATOM
4251
OH2
WAT
W
261
−18.067
−21.798
24.685
1
37.58
W


ATOM
4252
OH2
WAT
W
262
−32.052
0.596
−17.14
1
42.21
W


ATOM
4253
OH2
WAT
W
263
−37.602
−6.314
−21.983
1
23.91
W


ATOM
4254
OH2
WAT
W
264
−25.681
17.449
−3.446
1
36.89
W


ATOM
4255
OH2
WAT
W
265
7.708
−21.023
28.678
1
35.8
W


ATOM
4256
OH2
WAT
W
266
−47.298
3.593
−23.244
1
31.13
W


ATOM
4257
OH2
WAT
W
267
−32.028
11.145
−13.252
1
29.35
W


ATOM
4258
OH2
WAT
W
268
−20.602
−8.421
15.21
1
38.28
W


ATOM
4259
OH2
WAT
W
269
−23.157
3.393
15.329
1
42.06
W


ATOM
4260
OH2
WAT
W
270
−17.256
−30.56
30.887
1
44.11
W


ATOM
4261
OH2
WAT
W
271
−54.012
4.538
10.175
1
33.57
W


ATOM
4262
OH2
WAT
W
272
−51.255
−0.807
−33.214
1
35.29
W


ATOM
4263
OH2
WAT
W
273
−53.7
3.991
−28.662
1
38.54
W


ATOM
4264
OH2
WAT
W
274
−49.205
−11.226
−31.783
1
26.88
W


ATOM
4265
OH2
WAT
W
275
−3.226
−31.205
6.922
1
28.58
W


ATOM
4266
OH2
WAT
W
276
−9.21
0.335
11.738
1
35.26
W


ATOM
4267
OH2
WAT
W
277
−56.373
−3.491
−6.947
1
47.25
W


ATOM
4268
OH2
WAT
W
278
−63.906
−6.002
−29.092
1
38.3
W


ATOM
4269
OH2
WAT
W
279
−6.353
−16.779
6.373
1
35.03
W


ATOM
4270
OH2
WAT
W
280
−42.524
−7.674
−1.884
1
40.79
W


ATOM
4271
OH2
WAT
W
281
−50.135
−2.304
3.398
1
43.58
W


ATOM
4272
OH2
WAT
W
282
−16.141
10.089
−2.56
1
33.87
W


ATOM
4273
OH2
WAT
W
284
−21.282
9.545
−9.077
1
31.69
W


ATOM
4274
OH2
WAT
W
285
−50.552
0.916
−1.554
1
39.26
W


ATOM
4275
OH2
WAT
W
286
−54.679
10.607
−23.413
1
40.45
W


ATOM
4276
OH2
WAT
W
287
11.478
−7.558
33.467
1
26.36
W


ATOM
4277
OH2
WAT
W
288
−16.509
−20.371
17.394
1
32.57
W


ATOM
4278
OH2
WAT
W
289
−63.008
−1.711
−32.729
1
36.94
W


ATOM
4279
OH2
WAT
W
291
3.093
−5.855
28.07
1
38.84
W


ATOM
4280
OH2
WAT
W
292
−3.959
9.799
8.543
1
40.85
W


ATOM
4281
OH2
WAT
W
293
−9.428
−2.653
30.598
1
31
W


ATOM
4282
OH2
WAT
W
294
−48.06
−6.47
−2.302
1
37.85
W


ATOM
4283
OH2
WAT
W
295
−36.722
8.58
9.827
1
35.28
W


ATOM
4284
OH2
WAT
W
296
2.714
2.572
18.405
1
34.9
W


ATOM
4285
OH2
WAT
W
297
−25.826
6.21
20.872
1
37.94
W


ATOM
4286
OH2
WAT
W
298
−15.5
−21.562
44.837
1
35.67
W


ATOM
4287
OH2
WAT
W
301
−57.718
−5.868
−9.553
1
48.56
W


ATOM
4288
OH2
WAT
W
302
−5.575
−30.053
39.098
1
35.33
W


ATOM
4289
OH2
WAT
W
303
−14.494
−3.265
16.496
1
31.26
W


ATOM
4290
OH2
WAT
W
304
−42.319
−24.997
3.576
1
38.82
W


ATOM
4291
OH2
WAT
W
305
−3.526
−28.656
40.417
1
38.42
W


ATOM
4292
OH2
WAT
W
306
−8.2
−39.856
24.829
1
31.6
W


ATOM
4293
OH2
WAT
W
307
−45.478
−15.91
4.085
1
30.64
W


ATOM
4294
OH2
WAT
W
308
−14.914
6.149
19.737
1
44.58
W


ATOM
4295
OH2
WAT
W
309
−7.269
−5.334
36.669
1
33.28
W


ATOM
4296
OH2
WAT
W
311
−26.621
−26.002
21.739
1
28.54
W


ATOM
4297
OH2
WAT
W
312
−36.374
3.519
−13.879
1
41.18
W


ATOM
4298
OH2
WAT
W
313
−24.688
−24.74
15.049
1
39.88
W


ATOM
4299
OH2
WAT
W
314
−40.996
11.854
4.511
1
31.76
W


ATOM
4300
OH2
WAT
W
315
−24.136
−18.054
39.092
1
41.4
W









    Coordinates from minimization and B-factor refinement
    Refinement resolution: 50-2.50 A
    Starting r=0.2080 free_r=0.2831
    Final r=0.2046 free_r=0.2817
    Rmsd bonds=0.006567 rmsd angles=1.44990
    B rmsd for bonded mainchain atoms=1.833 target=2.0
    B rmsd for bonded sidechain atoms=2.568 target=2.5
    B rmsd for angle mainchain atoms=2.972 target=2.5
    B rmsd for angle sidechain atoms=3.689 target=3.0
    Target=mlf final wa=4.28786
    Final rweight=0.0733 (with wa=4.28786)
    Cycles=1 coordinate steps=50 B-factor steps=25
    Sg=P2(1)2(1)2(1) a=52.4 b=83.3 c=110.6 alpha=90 beta=90 gamma=90
    Topology file 1: CNS_TOPPAR:protein.top
    Topology file 2: CNS_TOPPAR:dna-ma.top
    Topology file 3: CNS_TOPPAR:water.top
    Topology file 4: CNS_TOPPAR:ion.top
    Parameter file 1: CNS_TOPPAR:protein—rep.param
    Parameter file 2: CNS_TOPPAR:dna-ma—rep.param
    Parameter file 3: CNS_TOPPAR:water_rep.param
    Parameter file 4: CNS_TOPPAR:ion.param
    Molecular structure file: gen.d.mtf
    Input coordinates: gen.d.pdb
    Reflection file=del_cv.hkl
    ncs=none
    B-correction resolution: 6.0-2.50
    Initial B-factor correction applied to fobs:
    B11=−5.213 B22=2.248 B33=2.965
    B12=0.000 B13=0.000 B23=0.000
    B-factor correction applied to coordinate array B: 0.640
    Bulk solvent: density level=0.356424 e/A^3, B-factor=44.9161 A^2
    Reflections with |Fobs|/sigma_F<0.0 rejected
    Reflections with |Fobs|>10000*rms(Fobs) rejected
    Theoretical total number of refl. in resol. range: 17370 (100.0%)
    Number of unobserved reflections (no entry or |F|=0): 618 (3.6%)
    Number of reflections rejected: 0 (0.0%)
    Total number of reflections used: 16752 (96.4%)
    Number of reflections in working set: 15935 (91.7%)
    Number of reflections in test set: 817 (4.7%)
    CRYST1 52.400 83.300 110.600 90.00 90.00 90.00 P21 21 21

## Claims

1. A crystal comprising domain III of West Nile Virus (WNV) Envelope protein (E protein) in association with an E16 Fab fragment in crystalline form, wherein the domain III of WNV E protein consists of SEQ ID NO: 1, wherein the E16 Fab fragment consists of SEQ ID NOS: 2 and 3, wherein said crystal forms in space group P212121 with unit cell dimensions a =52.4 ±0.2 Å, b =83.3 ±0.2 Å, and c =110.6 ±0.2 Å, and wherein α=β=γ=90 °.