# Patent

## Title
Crystal structure of monoacylglycerol lipase (MGLL)

## Abstract

A number of soluble engineered forms of MGLL that are suitable for high-throughput screening and protein crystallization, as well as a crystallized form of monoacylglycerol lipase protein (MGLL) and descriptions of the X-ray diffraction patterns are disclosed. The engineered constructs of MGLL permit the expression and purification of protein suitable for crystallography or high-throughput screening and identification of ligands, which can function as active agents to MGLL. The X-ray diffraction patterns allow the three dimensional structure of MGLL to be determined at atomic resolution so that ligand binding sites on MGLL can be identified and the interactions of ligands with MGLL amino acid residues can be modeled. Models prepared using such maps permit the design of ligands which can function as active agents which include, but are not limited to, those that function as inhibitors of MGLL.

## Background
TECHNICAL FIELD
The present invention generally pertains to the fields of molecular biology, protein purification, high-throughput screening, protein crystallization, X-ray diffraction analysis, three-dimensional structural determination, molecular modelling, and structure based rational drug design. The present invention provides a number of soluble engineered forms of MGLL that are suitable for high-throughput screening and protein crystallization, as well as a crystallized form of monoacylglycerol lipase protein (MGLL) and descriptions of the X-ray diffraction patterns.
SEQUENCE LISTING
The instant application contains a Sequence Listing which has been submitted in ASCII format via EFS-Web and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Oct. 11, 2010, is named PRD297US.txt and is 25,305 bytes in size.
The forms of MGLL provided by the present invention permit the expression and purification of protein suitable for high-throughput screening and crystallography. Thus forms of MGLL of the present invention have applications to the screening of MGLL to identify active agents which include, but are not limited to, those that find use as inhibitors of MGLL.
The X-ray diffraction patterns of the crystal of the present invention are of sufficient resolution so that the three-dimensional structure of MGLL can be determined at atomic resolution, ligand-binding sites on MGLL can be identified, and the interactions of ligands with amino acid residues of MGLL can be modeled. The high resolution maps provided by the present invention and the models prepared using such maps permit the design of ligands which can function as active agents. Thus, the three-dimensional structure of MGLL of the present invention has applications to the design of active agents, which include, but are not limited to, those that find use as inhibitors of MGLL.
BACKGROUND OF THE INVENTION
Various publications, which may include patents, published applications, technical articles and scholarly articles, are cited throughout the specification in parentheses, and full citations of each may be found at the end of the specification. Each of these cited publications is incorporated by reference herein, in its entirety.
Δ9-Tetrahydrocannabinol (THC) is the main psychoactive substance found in the cannabis plant. THC activates two distinct G protein-coupled receptors, cannabinoid 1 receptor (CB1) and cannabinoid 2 receptor (CB2) (Matsuda et al. 1990; Munro et al. 1993). CB1 is primarily expressed in the central nervous system (CNS) (Hohmann and Herkenham 1999; Farquhar-Smith et al. 2000; Rice et al. 2002; Walczak et al. 2005). CB2 expression, however, seems to be restricted to only peripheral tissues (Munro et al. 1993; Galiegue et al. 1995).
CNS mediated analgesic effects of cannabinoids have been well documented, but there is also accumulating evidence suggesting that cannabinoids can produce antinociception through peripheral mechanisms involving CB1 or CB2 (Hohmann 2002) (Malan et al. 2002). Richardson et al. demonstrated that cannabinoid antihyperalgesic effects were predominantly mediated by CB1 (Richardson et al. 1998; Richardson 2000). Hanus et al. showed that intraperitoneal injection of a CB2 selective agonist could suppress the late-phase response in the formalin test (Hanus et al. 1999). It was also shown that a CB2 selective agonist could attenuate thermal nociception and hyperalgesia (Malan et al. 2001; Malan et al. 2002; Quartilho et al. 2003) or suppress hyperalgesia evoked by intradermal administration of capsaicin (Hohmann et al. 2004). Ibrahim et al. showed that activation of CB2 with a selective CB2 agonist inhibited experimental neuropathic pain (Ibrahim et al. 2006). Taken together, the accumulating evidence clearly suggests great potential therapeutic value in targeting CB2 as a peripheral target for the treatment of pain. It should be noted that a significant advantage of this approach is that it would preclude unwanted CNS side effects caused by targeting CB1.
An arachidonic acid derivative, 2-arachidonyl glycerol (2-AG), is one of the two major and most well studied endogenous ligands for CB1 and CB2 (Gonsiorek et al. 2000). It has been shown that 2-AG acts as a potent and full-efficacy agonist of CB2 (Gonsiorek et al. 2000; Sugiura et al. 2000; Maresz et al. 2005) and that 2-AG is primarily hydrolysed by monoacylglycerol lipase (MGLL) (Dinh et al. 2002; Dinh et al. 2004; Saario et al. 2004). A non-competitive MGLL inhibitor that blocked 2-AG hydrolysis was found to enhance 2-AG levels and antinociception in stress models (Hohmann et al. 2005; Makara et al. 2005). It was also demonstrated that local administration of either 2-AG or a selective MGLL inhibitor induced a dose-dependent antinociceptive effect in an inflammatory pain model. Furthermore, local administration of the selective MGLL inhibitor in combination with 2-AG produced an additive antinociceptive effect (Guindon et al. 2007). Thus selective inhibition of MGLL may provide a novel therapeutic approach for the treatment of pain. Hitting this target, however, is inconceivable without good knowledge of the enzyme (Vandevoorde and Lambert 2005).
Lipases are lipolytic enzymes that have been differentiated from carboxylesterases by the fact that lipases have improved kinetics of hydrolysis for emulsions formed in oversaturated solutions. Carboxylesterases have been shown to have maximal activity using solutions of short-chain esters, with half-maximal activity at substrate concentrations far below the solubility limit. Exceeding the solubility limit was shown to have no effect on carboxylesterase activity. Lipases, on the other hand, were shown to have maximal activity using emulsified substrates, with half-maximal activity at substrate concentrations near the solubility limit (Chahinian et al. 2002). Early work with porcine pancreatic lipase showed that activity was low using a solution of ester substrates and abruptly increased as soon as an emulsion was formed. It was speculated that the porcine pancreatic lipase was activated by a conformational change of the enzyme as it bound to its water-insoluble substrate. The work with porcine pancreatic lipase was reviewed by Nini et al. (Nini et al. 2001).
In general, lipases share a similar α/β hydrolase fold with a catalytic Ser-His-Asp triad buried beneath a flexible cap-domain which is also referred to as a “lid” or “flap” (Brady et al. 1990; Winkler et al. 1990; Schrag et al. 1991). Although there is little conservation in the primary sequence of the cap-domain, it is generally formed of loops and one or more amphipathic helices. The cap-domains of human and dog gastric lipase are composed of intricate mixtures of 8 helices, turns, and random coils (Roussel et al. 1999; Roussel et al. 2002). In the crystal structure of human pancreatic lipase the lid adopts a helix-turn helix motif composed of two short amphipathic helices (van Tilbeurgh et al. 1992).
It has long been proposed that higher lipase activity for substrates presented as multimolecular aggregates (interfacial activation) is due to a conformational change in the cap-domain. It has also been proposed that changes from a closed to an open conformation of the lid is triggered by interaction with the substrate or lipid membrane (Brzozowski et al. 1991; van den Berg et al. 1995; van den Berg et al. 1995; Nini et al. 2001). Several other reports have also indicated that the loop covering the active site mediates lipase substrate specificity (Dugi et al. 1992; Dugi et al. 1995). It was demonstrated that movement of the helical lid results in a change in the hydrophobic-hydrophilic balance of the exterior surface of the lipase with the hydrophobic side of the lid becoming completely exposed in the active enzyme (Faustinella et al. 1992). Some lipases, such as guinea pig pancreatic lipase and bile salt-activated lipase, do not have a lid domain. Their active sites are freely accessible to solvent. As expected, based on the lack of the a cap-domain, these lipases are not activated by a lipid/water interface (Wang et al. 1997) (Carriere et al. 1997).
Although much has been learned about the structure of lipases through determination of three-dimensional structures of several microbial lipases and mammalian lipases, the three-dimensional structure of MGLL is unknown and its mechanism of action is not well understood. Furthermore, MGLL shows very little sequence similarity with other mammalian lipases and is unique among lipases in having monoglycerides as its only substrates. MGLL seems to be only distantly related to microbial proteins that include esterases, lysophospholipases, and haloperoxidases (Karlsson et al. 1997). A virtual molecular model of MGLL was built based on the crystal structure of chloroperoxidase (Saario et al. 2005; Saario et al. 2006). The model shows an alpha beta hydrolase fold with a lid domain comprised of four helices. The model, however, is only a virtual model and gives little insight into the actual mechanism of action of MGLL.
A crystal structure of MGLL would greatly facilitate the effort to discover MGLL selective inhibitors. A potential problem for crystallization experiments with MGLL is that detergents have been required to purify and stabilize MGLL in solution for both recombinant MGLL and MGLL from endogenous sources. (Tornqvist and Belfrage 1976; Somma-Delpero et al. 1995; Karlsson et al. 2000). Without detergent the purified MGLL protein was prone to aggregation. Crystallizing a detergent-solubilized protein into a structure of sufficient regularity to enable high-resolution X-ray crystallography can be problematic because well-ordered protein crystals can be difficult to obtain (U.S. Pat. No. 6,172,262B1).
The present invention provides a number of soluble engineered forms of monoacylglycerol lipase protein (MGLL) that do not require detergent for purification. The forms of MGLL provided by the present invention permit the expression and purification of protein suitable for identifying active agents in high-throughput screening and for crystallography. The present invention also provides a crystallized form of MGLL and descriptions of the X-ray diffraction patterns. Selective point mutations of hydrophobic residues in the cap-domain of MGLL generated soluble protein that did not require detergent for purification and stability. The protein displayed monomeric behaviour by gel filtration and was suitable for crystallization and high-throughput screening. In addition, selective mutation of surface lysine residues produced protein that generated crystals of improved quality. The crystal structure of MGLL was determined at atomic resolution. The forms of MGLL provide protein that can be used to identify inhibitors in high-throughput screens and the crystal structure of MGLL provides an important tool for structure-based drug design of MGLL inhibitors.
SUMMARY OF THE INVENTION
According to a first aspect of the present invention, there is provided a composition comprising a form of MGLL, or a fragment, or target structural motif or derivative thereof, wherein one or more hydrophobic residues of the cap-domain is mutated to improve solubility.
The present invention also provides a composition comprising a form of MGLL, or a fragment, or target structural motif or derivative thereof, wherein one or more hydrophobic Leucine residues of the cap-domain is mutated to improve solubility.
The present invention further provides a composition comprising a form of MGLL comprising one or more mutated hydrophobic Leucine residues of the cap-domain, wherein said one or more mutated hydrophobic Leucine residues of the cap-domain is selected from the group consisting of Leucine 162, Leucine 167, Leucine 169, Leucine 171, Leucine 174, Leucine 176, and Leucine 184, numbering based on the reference sequence for human MGLL Isoform 2 (SEQ ID NO: 1).
The present invention provides a composition comprising a form of MGLL, comprising one or more mutated hydrophobic residues of the cap-domain, wherein said one or more hydrophobic residues of the cap-domain is mutated to Serine, Glutamine, or Arginine.
In another aspect of the invention, the present invention includes a form of MGLL comprising one or more mutated hydrophobic Leucine residues of the cap-domain further comprising a Lysine mutated to an Alanine.
The present invention also includes a form of MGLL comprising one or more mutated hydrophobic Leucine residues of the cap-domain further comprising a Lysine mutated to an Alanine, wherein said Lysine residue is selected from the group consisting of Lysine 36, Lysine 160, Lysine 165, Lysine 188, Lysine 206, Lysine 226, Lysine 259 and Lysine 269, numbering based on the reference sequence for human MGLL Isoform 2.
The present invention further includes a method of identifying an agent that binds to the forms of MGLL of the present invention, comprising the steps of contacting the form of MGLL with the agent; measuring the binding of the agent to the form of MGLL; and, determining that the agent binds to the form of MGLL; thereby identifying an agent that binds to the form of MGLL.
In a preferred embodiment, the present invention includes a method of identifying an agent that binds to the forms of MGLL, wherein the form of MGLL has an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7.
In another preferred embodiment, the present invention includes a method of identifying an agent that binds to the forms of MGLL, wherein the binding is measured by the thermal stability of the form of MGLL.
The present invention further includes a method of identifying an agent that inhibits the activity of the forms of MGLL of the present invention comprising the steps of contacting the form of MGLL with the agent; measuring the biological activity of the form of MGLL in the presence of the agent; measuring the biological activity of the form of MGLL without the agent; and, comparing the biological activity of the form of MGLL measured in the presence of the agent and without the agent; thereby identifying the agent that decreases the biological activity the biological activity of the form of MGLL, when the activity measured in the presence of the agent is less than the activity measured without the agent.
In a preferred embodiment, the present invention includes a method of identifying an agent that inhibits the activity of the forms of MGLL, wherein the form of MGLL has an amino acid sequence selected from the group consisting of SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, and SEQ ID NO: 7.
In another preferred embodiment, the present invention includes a method of identifying an agent that inhibits the activity of the forms of MGLL of the present invention, wherein the biological activity is measured with an enzyme assay.
The present invention further includes methods of producing and using three-dimensional structure information derived from the crystal structure of monoacylglycerol lipase protein (MGLL).
The present invention also includes specific crystallization conditions to obtain crystals of the inhibitor-MGLL complex. The crystals are subsequently used to obtain a 3-dimensional structure of the complex using X-ray crystallography. The obtained data is used for rational drug discovery with the aim to design compounds that are inhibitors of MGLL.
The present invention includes a crystal comprising monoacylglycerol lipase (MGLL), or a fragment, or target structural motif or derivative thereof, and a ligand, wherein the ligand is a small molecule inhibitor. In another embodiment, the crystal has a spacegroup of C2221.
In another aspect of the invention, the present invention includes a crystal comprising a form of MGLL which comprises a peptide having at least 95% sequence identity to SEQ ID NO: 7.
In another aspect of the invention, the invention includes a computer system comprising: (a) a database containing information on the three dimensional structure of a crystal comprising MGLL, or a fragment or a target structural motif or derivative thereof, and a ligand, wherein the ligand is a small molecule inhibitor, stored on a computer readable storage medium; and, (b) a user interface to view the information.
The present invention also includes a method of evaluating the potential of an agent to associate with MGLL comprising: (a) exposing MGLL to the agent; and (b) detecting the association of said agent to MGLL amino acid residues SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 thereby evaluating the potential of the agent.
The invention further includes a method of evaluating the potential of an agent to associate with the peptide having the sequence of SEQ ID NO: 7, comprising: (a) exposing SEQ ID NO: 7 to the agent; and (b) detecting the level of association of the agent to SEQ ID NO: 7, thereby evaluating the potential of the agent.
Further included in the present invention is a method of identifying a potential agonist or antagonist against monoacylglycerol lipase comprising: (a) employing the three dimensional structure of MGLL cocrystallized with a small molecule inhibitor to design or select said potential agonist or antagonist.
The invention comprises a method of locating the attachment site of an inhibitor to monoacylglycerol lipase, comprising: (a) obtaining X-ray diffraction data for a crystal of MGLL; (b) obtaining X-ray diffraction data for a complex of MGLL and an inhibitor; (c) subtracting the X-ray diffraction data obtained in step (a) from the X-ray diffraction data obtained in step (b) to obtain the difference in the X-ray diffraction data; (d) obtaining phases that correspond to X-ray diffraction data obtained in step (a); (e) utilizing the phases obtained in step (d) and the difference in the X-ray diffraction data obtained in step (c) to compute a difference Fourier image of the inhibitor; and, (f) locating the attachment site of the inhibitor to MGLL based on the computations obtained in step (e).
The present invention further comprises a method of obtaining a modified inhibitor comprising: (a) obtaining a crystal comprising MGLL and an inhibitor; (b) obtaining the atomic coordinates of the crystal; (c) using the atomic coordinates and one or more molecular modelling techniques to determine how to modify the interaction of the inhibitor with MGLL; and, (d) modifying the inhibitor based on the determinations obtained in step (c) to produce a modified inhibitor.
In another aspect of the invention, the invention includes an isolated protein fragment comprising a binding pocket or active site defined by structure coordinates of MGLL amino acid residues SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279.
In another aspect of the invention, the invention includes an isolated nucleic acid molecule encoding the fragment which comprises a binding pocket or active site defined by structure coordinates of MGLL amino acid residues SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279. In another aspect of the invention, the invention includes a method of screening for an agent that associates with MGLL, comprising: (a) exposing a protein molecule fragment to the agent; and (b) detecting the level of association of the agent to the fragment. In another aspect of the invention, the invention includes a kit comprising a protein molecule fragment.
The invention additionally comprises a method for the production of a crystal complex comprising a MGLL polypeptide-ligand comprising: (a) contacting the MGLL polypeptide with said ligand in a suitable solution comprising PEG MME 5K, Na Citrate pH5.5 and n-Octyl-Beta-D-Glucopyranoside; and b) crystallizing said resulting complex of MGLL polypeptide-ligand from said solution.
The invention further includes a method for the production of a crystal comprising MGLL and a ligand wherein the ligand is a small molecule inhibitor comprising crystallizing a peptide comprising SEQ ID NO: 7 with a potential inhibitor.
The invention includes a method for identifying a potential inhibitor of monoacylglycerol lipase comprising: a) using a three dimensional structure of MGLL as defined by atomic coordinates according to Table 5; b) replacing one or more MGLL amino acids selected from SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 in said three-dimensional structure with a different amino acid to produce a modified MGLL; c) using said three-dimensional structure to design or select said potential inhibitor; d) synthesizing said potential inhibitor; and, e) contacting said potential inhibitor with said modified MGLL in the presence of a substrate to test the ability of said potential inhibitor to inhibit MGLL or said modified MGLL. Also included in the invention is an inhibitor identified by the method.



BRIEF DESCRIPTION OF THE DRAWINGS
A preferred embodiment of the present invention will now be described, by way of an example only, with reference to the accompanying drawings wherein:
FIG. 1: A. Shown is SEQ ID NO: 1, the amino acid sequence of human monoglyceride lipase isoform 2 (Karlsson et al. 2001), accession NP—001003794, version NP—001003794.1, GI:51242953. B. Shown is SEQ ID NO: 2, the amino acid sequence of human monoglyceride lipase isoform 1 (Wall et al. 1997), Accession NP—009214, Version NP—009214.1, GI:6005786. C. Shown is the sequence alignment for SEQ ID NO: 1 and residues 11-314 of SEQ ID NO: 2, which are human monoglyceride lipase isoform 2 and isoform 1, respectively. Sequence alignment was done with the online BLAST 2 SEQUENCES software (Tatusova and Madden 1999).
FIG. 2: A. Shown is the sequence alignment of human MGLL isoform 2 (SEQ ID NO: 1) and crystallized RsbQ (SEQ ID NO: 11) (RsbQ, PDB 1WOM). Sequence alignment was done with the ClustalW software. Homologous residues are shown in black. Identical residues are shown in green. Position of helices and β sheets are indicated by red stars and green columns, respectively. The active site residues are highlighted in red. Leu 169 and Leu 176 are indicated by blue arrows. B. Shown is ribbon diagram representation of a homology model of human MGLL isoform 2 based on RsbQ (Pdb 1bro). Leu 169 and Leu 176 are shown in magenta. The active site residues are shown in green. C. Shown is surface representation of a homology model of human MGLL isoform 2 based on RsbQ (Pdb 1bro). Leu 169 and Leu 176 are shown in magenta. The active site residues are shown in green.
FIG. 3: A. Shown is SEQ ID NO: 3, the amino acid sequence of the construct used to express wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3). The N-terminal His tag is shown in non-capitalized text and the TEV cleavage site is shown in bold non-capitalized text with an arrow over the site of cleavage. B. Shown is SEQ ID NO: 4, the amino acid sequence of the construct used to express a form of mut-MGLL (hMGLL 1-303 L169S, L176S). The N-terminal His tag is shown in non-capitalized text, the TEV cleavage site is shown in bold non-capitalized text with an arrow over the site of cleavage, and the mutations are shown in bold non-capitalized text. C. Shown is SEQ ID NO: 5, the amino acid sequence of mut-MGLL (hMGLL 1-303 L169S, L176S) after TEV cleavage of the N-terminal His tag. The one amino acid from the TEV cleavage site that remains after TEV cleavage is shown in bold non-capitalized text and the mutations are shown in bold non-capitalized text. D. Shown is SEQ ID NO: 6, the amino acid sequence of the construct used to express a form of mut-MGLL (hMGLL 1-303 L169S, L176S, K36A). The N-terminal His tag is shown in non-capitalized text, the TEV cleavage site is shown in bold non-capitalized text with an arrow over the site of cleavage, and the mutations are shown in bold non-capitalized text. E. Shown is SEQ ID NO: 7, the amino acid sequence of mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) after TEV cleavage of the N-terminal His tag. The one amino acid from the TEV cleavage site that remains after TEV cleavage is shown in bold non-capitalized text and the mutations are shown in bold non-capitalized text.
FIG. 4: A. Shown are size exclusion elution profiles for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) (magenta dotted lines) and TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5) (blue solid line) purified in the absence of detergent showing 100% aggregation for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and 90% monomer for TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5). B. Shown is circular dichroism structural analysis of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5). Far UV scans from 200 to 260 nm are shown in (i) and temperature melts from 25 to 80° C. monitored at 210 nm are shown in (ii).
FIG. 5: Shown are duplicate Michaelis-Menten curves for the hydrolysis of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5).
FIG. 6: Shown are thermal shift data of the melting transitions for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) (green line) and TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5) (red line). The midpoint of the melting transition was 58° C. for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and 56.7° C. for TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5).
FIG. 7: A. Shown is the structure of Compound 1. B. Shown is the structure of Compound 2.
FIG. 8: Shown is a ribbon diagram of the structure of the complex of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) and Compound 2. Color-coding is according to secondary structure (α-helices: magenta, β-sheets: yellow). The ligand (Compound 2) is drawn in ball-and-stick representation (green). The protein adopts a typical α/β hydrolase fold comprised of 8 β-sheets, with β2 being antiparallel to the other sheets. The inhibitor is located in the active site, which is capped by loops connecting α4 to α6.
FIG. 9: Shown is a surface representation of the structure for the complex of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) and Compound 2.
FIG. 10: A. Shown is a ribbon diagram of MGLL with the cap-domain (residues 145-206) colored cyan to illustrate the location of the cap-domain. B. Shown is an overlay of the ribbon diagram of MGLL (orange) onto Bromoperoxidase A1 (salmon) and Chloroperoxidase L (green); C. Shown is an overlay of the ribbon diagram of MGLL (orange) onto P. putida Esterase (grey) and Gamma Lactamase (yellow). The cap-domain was omitted for clarity to show the good alignment between the core α/β-hydrolase in all structures. D. Shown is a ribbon diagram of the MGLL cap-domain. Superposition of different cap-domains highlighting the different arrangement between MGLL and other hydrolases. E. Shown is an overlay of the ribbon diagrams of the cap-domains of Bromoperoxidase A1 (salmon) and Chloroperoxidase L (green); F. Shown is an overlay of the ribbon diagrams of the cap-domains of P. putida Esterase (grey) and Gamma Lactamase (yellow).
FIG. 11: Shown is a ribbon diagram of MGLL showing the location of mutations L169S and L176S in the cap-domain that prevented protein aggregation and K36A on the loop between β2 and β4 that would make interactions with a symmetry mate if present in the wild-type protein.



Table 1: Shown is a table of the forms of MGLL of the present invention and the purification yield in mg/liter.
Table 2: Shown is a table of the kinetic constants of the various MGLL constructs using 4MC-B or C-A as substrates. Values for the 4MC-B substrate are the average of 2 or 4 separate assays. The kcat/KM values for the C-A substrate are the average values for the hydrolysis of five different substrate concentrations at [S]<<KM.
Table 3: Shown are the data collection and refinement statistics for the complex of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) and Compound 2.
Table 4: Shown is are the superposition statistics for selected α/β hydrolases without the cap-domain superimposed onto MGLL without the cap-domain
Table 5: Shown are the coordinates for the complex of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 10) and Compound 2.
DEFINITIONS
As is generally the case in biotechnology and chemistry, the description of the present invention has required the use of a number of terms of art. Although it is not practical to do so exhaustively, definitions for some of these terms are provided here for ease of reference. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Definitions for other terms may also appear elsewhere herein. However, the definitions provided here and elsewhere herein should always be considered in determining the intended scope and meaning of the defined terms. Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods and materials are described.
The term “comprising” means “including principally, but not necessarily solely”. Furthermore, variations of the word “comprising”, such as “comprise” and “comprises”, have correspondingly varied meanings.
As used herein, the terms “containing”, “having” and “including” are used in their open, non-limiting sense.
As used herein, “sequence” means the linear order in which monomers occur in a polymer, for example, the order of amino acids in a polypeptide or the order of nucleotides in a polynucleotide.
The terms “polypeptide,” “protein,” and “peptide” are used herein interchangeably to refer to amino acid chains in which the amino acid residues are linked by peptide bonds or modified peptide bonds. The amino acid chains can be of any length of greater than two amino acids. Unless otherwise specified, the terms “polypeptide,” “protein,” and “peptide” also encompass various modified forms thereof. Such modified forms may be naturally occurring modified forms or chemically modified forms. Examples of modified forms include, but are not limited to, glycosylated forms, phosphorylated forms, myristoylated forms, palmitoylated forms, ribosylated forms, acetylated forms, ubiquitinated forms, etc. Modifications also include intra-molecular crosslinking and covalent attachment to various moieties such as lipids, flavin, biotin, polyethylene glycol or derivatives thereof, etc. In addition, modifications may also include cyclization, branching and cross-linking. Further, amino acids other than the conventional twenty amino acids encoded by the codons of genes may also be included in a polypeptide.
As used herein, a protein or nucleic acid molecule is said to be “isolated” when the protein or nucleic acid molecule is substantially separated from contaminants from the source of the protein or nucleic acid.
As used herein, the term “native protein” refers to a protein comprising an amino acid sequence identical to that of a protein isolated from its natural source or organism.
As used herein, the term “amino acids” refers to the L-isomers of the naturally occurring amino acids. The naturally occurring amino acids are glycine, alanine, valine, leucine, isoleucine, serine, methionine, threonine, phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine, aspartic acid, asparagine, glutamic acid, glutamine, γ-carboxylglutamic acid, arginine, ornithine, and lysine. Unless specifically indicated, all amino acids are referred to in this application are in the L-form.
As used herein, the term “nonnatural amino acids” refers to amino acids that are not naturally found in proteins. For example, selenomethionine.
As used herein, the term “positively charged amino acid” includes any amino acids having a positively charged side chain under normal physiological conditions. Examples of positively charged naturally occurring amino acids are arginine, lysine, and histidine.
As used herein, the term “negatively charged amino acid” includes any amino acids having a negatively charged side chains under normal physiological conditions. Examples of negatively charged naturally occurring amino acids are aspartic acid and glutamic acid.
As used herein, the term “hydrophobic amino acid” includes any amino acids having an uncharged, nonpolar side chain that is relatively insoluble in water. Examples of naturally occurring hydrophobic amino acids are alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan, and methionine.
As used herein, the term “hydrophilic amino acid” refers to any amino acids having an uncharged, polar side chain that is relatively soluble in water. Examples of naturally occurring hydrophilic amino acids are serine, threonine, tyrosine, asparagine, glutamine and cysteine.
As used herein, “nucleic acid” is defined as RNA or DNA that encodes a protein or peptide as defined herein, or is complementary to nucleic acid sequence encoding such peptides, or hybridizes to such nucleic acid and remains stably bound to it under appropriate stringency conditions. Nucleic acid sequences can be composed of natural nucleotides of the following bases: thymidine, adenine, cytosine, guanine, and uracil; abbreviated T, A, C, G, and U, respectively, and/or synthetic analogs of the natural nucleotides.
The term “oligonucleotide” or “oligo” refers to a single-stranded DNA or RNA sequence of a relatively short length, for example, less than 100 residues long. For many methods, oligonucleotides of about 16-25 nucleotides in length are useful, although longer oligonucleotides of greater than about 25 nucleotides may sometimes be utilized. Some oligonucleotides can be used as “primers” for the synthesis of complimentary nucleic acid strands. For example, DNA primers can hybridize to a complimentary nucleic acid sequence to prime the synthesis of a complimentary DNA strand in reactions using DNA polymerases. Oligonucleotides are also useful for hybridization in several methods of nucleic acid detection, for example, in Northern blotting or in situ hybridization.
“Recombinant” refers to a nucleic acid, a protein encoded by a nucleic acid, a cell, or a viral particle, that has been modified using molecular biology techniques to something other than its natural state. For example, recombinant cells can contain nucleotide sequence that is not found within the native (non-recombinant) form of the cell or can express native genes that are otherwise abnormally, under-expressed, or not expressed at all. Recombinant cells can also contain genes found in the native form of the cell wherein the genes are modified and re-introduced into the cell by artificial means. The term also encompasses cells that contain an endogenous nucleic acid that has been modified without removing the nucleic acid from the cell; such modifications include those obtained, for example, by gene replacement, and site-specific mutation.
The term “high stringency” as used herein refers to the conditions under which two nucleic acids may be hybridized, and may include, for example, the concentration of salts and/or detergents in a solution, the temperature of a solution that is used during the hybridization of the two nucleic acids and time period of the hybridization. Accordingly, the term “high stringency” as used herein refers to conditions in a solution that are conducive to hybridization of two nucleic acids only where such nucleic acids share a high degree of complementarity. The degree of complementarity may include, but not be limited to, a range of from about 90% to 100%. Thus, “high stringency” conditions may involve, but are not limited to, the use of a varying temperature and a buffer comprising various concentrations of detergents, salts, and divalent cations.
As used herein, “vector” refers to a nucleic acid molecule into which a heterologous nucleic acid can be or is inserted. Some vectors can be introduced into a host cell allowing for replication of the vector or for expression of a protein that is encoded by the vector or construct. Vectors typically have selectable markers, for example, genes that encode proteins allowing for drug resistance, origins of replication sequences, and multiple cloning sites that allow for insertion of a heterologous sequence. Vectors are typically plasmid-based and are designated by a lower case “p” followed by a combination of letters and/or numbers. Starting plasmids disclosed herein are either commercially available, publicly available on an unrestricted basis, or can be constructed from available plasmids by application of procedures known in the art. Many plasmids and other cloning and expression vectors that can be used in accordance with the present invention are well-known and readily available to those of skill in the art. Moreover, those of skill readily may construct any number of other plasmids suitable for use in the invention. The properties, construction and use of such plasmids, as well as other vectors, in the present invention will be readily apparent to those of skill from the present disclosure.
As used herein, the term “activity” refers to an activity exerted by MGLL as determined in vivo or in vitro, according to standard techniques. Examples of such activity include, but are not limited to, direct activity such as the ability to bind to a ligand or an analog thereof, enzymatic activity, or functional changes of cell physiology that result from changes in activity.
The term “high-throughput assay” or “high-throughput screening” refers to assay designs that allow easy screening of multiple samples simultaneously and/or in rapid succession, and may include the capacity for robotic manipulation. Another desired feature of high-throughput assays is an assay design that is optimized to reduce reagent usage, or minimize the number of manipulations in order to achieve the analysis desired. Examples of high-throughput assay formats include, but are not limited to, formats that utilize 96-well, 384-well, and 1536-well plates, or “lab on a chip” microchannel chips used for liquid handling experiments. It is well known by those in the art that as miniaturization of plastic molds and liquid handling devices are advanced, or as improved assay devices are designed, greater numbers of samples can be processed using the forms of the present invention. Any high-throughput screening may be utilized to test new compounds, which are identified or designed for their ability to interact with MGLL. For general information on high-throughput screening see, for example, (Devlin (editor) 1998); and (U.S. Pat. No. 5,763,263).
By the term “selecting” or “select” compounds it is intended to encompass both (a) choosing compounds from a group previously unknown to be modulators of a protein complex or interacting protein members thereof, and (b) testing compounds that are known to be capable of binding, or modulating the functions and activities of, a protein complex or interacting protein members thereof. The compounds encompass numerous chemical classes, including but not limited to, small organic or inorganic compounds, natural or synthetic molecules, such as antibodies, proteins or fragments thereof, antisense nucleotides, interfering RNA (iRNA) and ribozymes, and derivatives, mimetics and analogs thereof. Preferably, they are small organic compounds, i.e., those having a molecular weight of no greater than 10,000 daltons, more preferably less than 5,000 daltons.
As used herein, the term “atomic coordinates” or “structure coordinates” refers to mathematical coordinates that describe the positions of atoms in crystals of MGLL in Protein Data Bank (PDB) format, including X, Y, Z and B, for each atom. The diffraction data obtained from the crystals are used to calculate an electron density map of the repeating unit of the crystal. The electron density maps may be used to establish the positions (i.e. coordinates X, Y and Z) of the individual atoms within the crystal. Those of skill in the art understand that a set of structure coordinates determined by X-ray crystallography is not without standard error. For the purpose of this invention, any set of structure coordinates for MGLL from any source having a root mean square deviation of non-hydrogen atoms of less than about 1.5 Å when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5 are considered substantially identical or homologous. In a more preferred embodiment, any set of structure coordinates for MGLL from any source having a root mean square deviation of non-hydrogen atoms of less than about 0.75 Å. when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5 are considered substantially identical or homologous.
The term “atom type” refers to the chemical element whose coordinates are measured. The abbreviations in column 3 of Table 5 identifies the element.
The terms “X,” “Y” and “Z” refer to the crystallographically-defined atomic position of the element measured with respect to the chosen crystallographic origin. The term “B” refers to a thermal factor that measures the mean variation of an atom's position with respect to its average position.
As used herein, the term “crystal” refers to any three-dimensional ordered array of molecules that diffracts X-rays.
As used herein, the term “carrier” in a composition refers to a diluent, adjuvant, excipient, or vehicle with which the product is mixed.
As used herein, the term “composition” refers to the combining of distinct elements or ingredients to form a whole. A composition comprises more than one element or ingredient. For the purposes of this invention, a composition will often, but not always comprise a carrier.
As used herein, “MGLL” is used to mean a protein obtained as a result of expression of monoacylglycerol lipase. Within the meaning of this term, it will be understood that human MGLL encompasses all proteins encoded by monoacylglycerol lipase, mutants thereof, conservative amino acid substitutions, alternative splice proteins thereof, and phosphorylated proteins thereof. Additionally, as used herein, it will be understood that the term “MGLL” includes monoacylglycerol lipase and homologues from other animals. As an example, MGLL includes the protein comprising SEQ ID NO: 7 and variants thereof comprising at least about 70% amino acid sequence identity to SEQ ID NO: 7, or preferably 80%, 85%, 90% and 95% sequence identity to SEQ ID NO: 7, or more preferably, at least about 95% or more sequence identity to SEQ ID NO: 7.
As used herein, the term “SAR,” an abbreviation for Structure-Activity Relationships, collectively refers to the structure-activity/structure property relationships pertaining to the relationship(s) between a compound's activity/properties and its chemical structure.
As used herein, the term “molecular structure” refers to the three dimensional arrangement of molecules of a particular compound or complex of molecules (e.g., the three dimensional structure of MGLL and ligands that interact with MGLL.
As used herein, the term “molecular modeling” refers to the use of computational methods, preferably computer assisted methods, to draw realistic models of what molecules look like and to make predictions about structure activity relationships of ligands. The methods used in molecular modeling range from molecular graphics to computational chemistry.
As used herein, the term “molecular model” refers to the three dimensional arrangement of the atoms of a molecule connected by covalent bonds or the three dimensional arrangement of the atoms of a complex comprising more than one molecule, e.g., a protein-ligand complex.
As used herein, the term “molecular graphics” refers to three dimensional (3D) representations of the molecules; for instance, a 3D representation produced using computer assisted computational methods.
As used herein, “computer readable medium” refers to any medium, which can be read and accessed directly by a computer. Such media include, but are not limited to: magnetic storage media, such as floppy discs, hard disc storage media, and magnetic tape; optical storage media such as optical discs or CD-ROM; electrical storage media such as RAM and ROM; and hybrids of these categories such as magnetic/optical storage media.
As used herein, “recorded” refers to a process for storing information on computer readable media. A skilled artisan can readily adopt any of the presently known methods for recording information on computer readable media to generate manufactures comprising an amino acid sequence and/or atomic coordinate/X-ray diffraction data information of the present invention.
As used herein, “a computer-based system” refers to the hardware means, software means, and data storage means used to analyze the sequence and/or X-ray diffraction data of the present invention. The minimum hardware means of the computer-based systems of the present invention comprises a central processing unit (CPU), input means, output means, and data storage means. A skilled artisan can readily appreciate which of the currently available computer-based systems are suitable for use in the present invention. A visualization device, such as a monitor, is optionally provided to visualize structure data.
As stated above, the computer-based systems of the present invention comprise a data storage means having stored therein sequence and/or atomic coordinate/X-ray diffraction data of the present invention and the necessary hardware means and software means for supporting and implementing an analysis means. As used herein, “data storage means” refers to memory which can store sequence or atomic coordinate/X-ray diffraction data of the present invention, or a memory access means which can access manufactures having recorded thereon the sequence or X-ray data of the present invention.
As used herein, “search means” or “analysis means” refers to one or more programs which are implemented on the computer-based system to compare a target sequence or target structural motif with the sequence or X-ray data stored within the data storage means. Search means are used to identify fragments or regions of a protein which match a particular target sequence or target motif. A variety of known algorithms are disclosed publicly and a variety of commercially available software for conducting search means are and can be used in the computer-based systems of the present invention. A skilled artisan can readily recognize that any one of the available algorithms or implementing software packages for conducting computer analyses can be adapted for use in the present computer-based systems.
As used herein, “a target structural motif,” or “target motif,” refers to any rationally selected sequence or combination of sequences in which the sequence(s) are chosen based on a three-dimensional configuration or electron density map which is formed upon the folding of the target motif. There are a variety of target motifs known in the art. Protein target motifs include, but are not limited to, enzymatic active sites, inhibitor binding sites, structural subdomains, epitopes, functional domains and signal sequences. Similar motifs are known for RNA. A variety of structural formats for the input and output means can be used to input and output the information in the computer-based systems of the present invention.
As used herein, the term “computational chemistry” refers to calculations of the physical and chemical properties of the molecules.
As used herein, the term “molecular replacement” refers to a method that involves generating a preliminary model of a crystal of MGLL whose coordinates are unknown, by orienting and positioning the said atomic coordinates described in the present invention so as best to account for the observed diffraction pattern of the unknown crystal. Phases can then be calculated from this model and combined with the observed amplitudes to give an approximate Fourier synthesis of the structure whose coordinates are unknown. (Rossmann 1972)
As used herein, the term “homolog” refers to the MGLL protein molecule or the nucleic acid molecule which encodes the protein, or a functional domain from said protein from a first source having at least about 70% or 75% sequence identity, or at least about 80% sequence identity, or more preferably at least about 85% sequence identity, or even more preferably at least about 90% sequence identity, and most preferably at least about 95%, 97% or 99% amino acid or nucleotide sequence identity, with the protein, encoding nucleic acid molecule or any functional domain thereof, from a second source. The second source may be a version of the molecule from the first source that has been genetically altered by any available means to change the primary amino acid or nucleotide sequence or may be from the same or a different species than that of the first source.
As used herein, the term “active site” refers to regions on MGLL or a structural motif of MGLL that are directly involved in the function or activity of human MGLL.
As used herein, the terms “binding site” or “binding pocket” refer to a region of human MGLL or a molecular complex comprising MGLL that, as a result of the primary amino acid sequence of human MGLL and/or its three-dimensional shape, favourably associates with another chemical entity or compound including ligands, cofactors, or inhibitors. For the purpose of this invention, any active site, binding site or binding pocket defined by a set of structure coordinates for MGLL or for a homolog of MGLL from any source having a root mean square deviation of non-hydrogen atoms of less than about 1.5 Å. when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5 are considered substantially identical or homologous. In a more preferred embodiment, any set of structure coordinates for MGLL or a homolog of MGLL from any source having a root mean square deviation of non-hydrogen atoms of less than about 0.75 Å. when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5 are considered substantially identical or homologous.
The term “root mean square deviation” means the square root of the arithmetic mean of the squares of the deviations from the mean.
As used herein, the term “hydrogen bond” refers to two hydrophilic atoms (either O or N), which share a hydrogen that is covalently bonded to only one atom, while interacting with the other.
As used herein, the term “hydrophobic interaction” refers to interactions made by two hydrophobic residues or atoms (such as C).
As used herein, the term “conjugated system” refers to more than two double bonds adjacent to each other, in which electrons are completely delocalized with the entire system. This also includes aromatic residues.
As used herein, the term “aromatic residue” refers to amino acids with side chains having a delocalized conjugated system. Examples of aromatic residues are phenylalanine, tryptophan, and tyrosine.
As used herein, the phrase “inhibiting the binding” refers to preventing or reducing the direct or indirect association of one or more molecules, peptides, proteins, enzymes, or receptors, or preventing or reducing the normal activity of one or more molecules, peptides, proteins, enzymes or receptors, e.g., preventing or reducing the direct or indirect association with human MGLL.
As used herein, the term “competitive inhibitor” refers to inhibitors that bind to human MGLL, thus directly competing with a substrate or ligand. Competitive inhibition may, in some instances, be reversed completely by increasing the substrate or ligand concentration.
As used herein, the term “uncompetitive inhibitor” refers to one that inhibits the functional activity of human MGLL by binding to a different site than does its substrate(s). As used herein, the term “non-competitive inhibitor” refers to one that can bind to either the free or bound form of MGLL. Those of skill in the art may identify inhibitors as competitive, uncompetitive, or non-competitive by computer fitting enzyme kinetic data using standard methods. See, for example, (Segel 1975)
As used herein, the term “R or S-isomer” refers to two possible stereoisomers of a chiral carbon according to the Cahn-Ingold-Prelog system adopted by International Union of Pure and Applied Chemistry (IUPAC). Each group attached to the chiral carbon is first assigned to a preference or priority a, b, c, or d on the basis of the atomic number of the atom that is directly attached to the chiral carbon. The group with the highest atomic number is given the highest preference a, the group with next highest atomic number is given the next highest preference b, and so on. The group with the lowest preference (d) is then directed away from the viewer. If the trace of a path from a to b to c is counter clockwise, the isomer is designated (S); in the opposite direction, clockwise, the isomer is designated (R).
As used herein, the term “stereoisomers” is a general term for all isomers of individual molecules that differ only in the orientation of their atoms in space. It includes enantiomers and isomers of compounds with more than one chiral center that are not mirror images of one another (diastereomers).
As used herein, the term “chiral center” refers to a carbon atom to which four different groups are attached.
As used herein, the term “enantiomer” or “enantiomeric” refers to a molecule that is nonsuperimposable on its mirror image and hence optically active wherein the enantiomer rotates the plane of polarized light in one direction and its mirror image rotates the plane of polarized light in the opposite direction.
As used herein, the term “racemic” refers to a mixture of equal parts of enantiomers and which is optically active.
As used herein, the term “resolution” refers to the separation or concentration or depletion of one of the two enantiomeric forms of a molecule. In the context of this application. The term “resolution” also refers to the amount of detail, which can be resolved by the diffraction experiment. Or in other terms, since the inherent disorder of a protein crystal diffraction pattern fades away at some diffraction angle thetamax, the corresponding distance dmin of the reciprocal lattices is determined by Bragg's law. In practice in protein crystallography it is usual to quote the nominal resolution of a protein electron density in terms of dmin, the minimum lattice distance to which data is included in the calculation of the map.
As used herein, the term “ligand” refers to any molecule, or chemical entity, which binds with or to MGLL, a subunit of MGLL, a domain of MGLL, a target structural motif of MGLL, or a fragment of MGLL. Thus, ligands include, but are not limited to, small molecule inhibitors, for example.
As used herein, the term “small molecule inhibitor” refers to compounds useful in the present invention having measurable MGLL inhibiting activity. In addition to small organic molecules, peptides, antibodies, cyclic peptides and peptidomimetics are contemplated as being useful in the disclosed methods. Preferred inhibitors are small molecules, preferably less than 10,000 daltons, and more preferably less than 5,000 daltons.
As used herein the terms “bind,” “binding,” “bond,” or “bonded” when used in reference to the association of atoms, molecules, or chemical groups, refer to any physical contact or association of two or more atoms, molecules, or chemical groups.
As used herein, the terms “covalent bond” or “valence bond” refer to a chemical bond between two atoms in a molecule created by the sharing of electrons, usually in pairs, by the bonded atoms.
As used herein, “noncovalent bond” refers to an interaction between atoms and/or molecules that does not involve the formation of a covalent bond between them.
DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS
It is to be understood at the outset, that the figures and examples provided herein are to exemplify, and not to limit the invention and its various embodiments.
The present invention includes a crystal comprising monoacylglycerol lipase (MGLL), or a fragment, or target structural motif or derivative thereof, and a ligand, wherein the ligand is a small molecule inhibitor. In one embodiment, the fragment or derivative thereof is a peptide comprising SEQ ID NO: 7
In another embodiment, the crystal has a spacegroup of C2221. In a different embodiment, the crystal effectively diffracts X-rays for determination of atomic coordinates to a resolution of at least about 3.2 Å. In a preferred embodiment, the ligand is in crystalline form. In a highly preferred embodiment, the ligand is the structure depicted in FIG. 7A or FIG. 7B, and, derivatives thereof.
The present invention also includes a crystal comprising MGLL, which comprises a peptide having at least 95% sequence identity to SEQ ID NO. 6. In a preferred embodiment, the crystal comprising SEQ ID NO: 7 comprises an atomic structure characterized by the coordinates of Table 5. In another preferred embodiment, the crystal comprises a unit cell selected from the group consisting of: a cell having dimensions of a=93.95, b=128.45, c=60.6.
In another aspect of the invention, the invention includes a computer system comprising: (a) a database containing information on the three dimensional structure of a crystal comprising MGLL, or a fragment or a target structural motif or derivative thereof, and a ligand, wherein the ligand is a small molecule inhibitor, stored on a computer readable storage medium; and, (b) a user interface to view the information. In one embodiment, the information comprises diffraction data obtained from a crystal comprising SEQ ID NO: 7.
In another embodiment, the information comprises an electron density map of a crystal form comprising SEQ ID NO: 7. In a different embodiment, the information comprises the structure coordinates of Table 5 or homologous structure coordinates comprising a root mean square deviation of non-hydrogen atoms of less than about 1.5 Å when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5. In a preferred embodiment, the information comprises structure coordinates comprising a root mean square deviation of non-hydrogen atoms of less than about 0.75 Å when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5. In a highly preferred embodiment, the information comprises the structure coordinates for amino acids SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 according to Table 5 or similar structure coordinates for said amino acids comprising a root mean square deviation of non-hydrogen atoms of less than about 1.5 Å to 0.75 Å when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5.
The present invention also includes a method of evaluating the potential of an agent to associate with MGLL comprising: (a) exposing MGLL to the agent; and (b) detecting the association of said agent to MGLL amino acid residues SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 thereby evaluating the potential. In one embodiment of the invention, the agent is a virtual compound. In another embodiment of the invention, step (a) comprises comparing the atomic structure of the compound to the three dimensional structure of MGLL. In a different embodiment, the comparing of step (a) comprises employing a computational means to perform a fitting operation between the compound and at least one binding site of MGLL. In a preferred embodiment, the binding site is defined by structure coordinates for amino acids SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 according to Table 5 or similar structure coordinates for said amino acids comprising a root mean square deviation of non-hydrogen atoms of less than about 1.5 Å when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5. In a highly preferred embodiment, step (a) comprise exposing the agent to crystalline SEQ ID NO: 7 and the detecting of step (b) comprises determining the three dimensional structure of the agent-SEQ ID NO: 7 complex.
The present invention includes a method of identifying a potential agonist or antagonist against MGLL comprising: (a) employing the three dimensional structure of MGLL cocrystallized with a small molecule inhibitor to design or select said potential agonist or antagonist. In one embodiment, the three dimensional structure corresponds to the atomic structure characterized by the coordinates of Table 5 or similar structure coordinates comprising a root mean square deviation of non-hydrogen atoms of less than about 1.5 Å when superimposed on the non-hydrogen atom positions of the corresponding atomic coordinates of Table 5. In a different embodiment, the method further comprises the steps of: (b) synthesizing the potential agonist or antagonist; and (c) contacting the potential agonist or antagonist with MGLL.
The instant invention comprises a method of locating the attachment site of an inhibitor to MGLL, comprising: (a) obtaining X-ray diffraction data for a crystal of MGLL; (b) obtaining X-ray diffraction data for a complex of MGLL and an inhibitor; (c) subtracting the X-ray diffraction data obtained in step (a) from the X-ray diffraction data obtained in step (b) to obtain the difference in the X-ray diffraction data; (d) obtaining phases that correspond to X-ray diffraction data obtained in step (a); (e) utilizing the phases obtained in step (d) and the difference in the X-ray diffraction data obtained in step (c) to compute a difference Fourier image of the inhibitor; and, (f) locating the attachment site of the inhibitor to MGLL based on the computations obtained in step (e).
The present invention further comprises a method of obtaining a modified inhibitor comprising: (a) obtaining a crystal comprising MGLL and an inhibitor; (b) obtaining the atomic coordinates of the crystal; (c) using the atomic coordinates and one or more molecular modeling techniques to determine how to modify the interaction of the inhibitor with MGLL; and, (d) modifying the inhibitor based on the determinations obtained in step (c) to produce a modified inhibitor. In one embodiment, the crystal comprises a peptide having a sequence comprising SEQ ID NO: 7. In a different embodiment, the one or more molecular modeling techniques are selected from the group consisting of graphic molecular modeling and computational chemistry. In a preferred embodiment, step (a) comprises detecting the interaction of the inhibitor to MGLL amino acid residues SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279. In another embodiment of the invention, the invention includes an MGLL inhibitor identified by this method.
In another aspect of the invention, the invention includes an isolated protein fragment comprising a binding pocket or active site defined by structure coordinates of MGLL amino acid residues SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279. In one embodiment, the isolated fragment is linked to a solid support.
In another aspect of the invention, the invention includes an isolated nucleic acid molecule encoding the fragment, which comprises a binding pocket or active site defined by structure coordinates of MGLL. In one embodiment, a vector comprises the nucleic acid molecule. In another embodiment, a host cell comprises the vector. In yet another aspect of the invention, the invention includes a method of producing a protein fragment, comprising culturing the host cell under conditions in which the fragment is expressed. In another aspect of the invention, the invention includes a method of screening for an agent that associates with MGLL, comprising: (a) exposing a protein molecule fragment to the agent; and (b) detecting the level of association of the agent to the fragment. In another aspect of the invention, the invention includes a kit comprising a protein molecule fragment.
In another aspect of the invention, the invention includes a method for the production of a crystal complex comprising an MGLL polypeptide-ligand comprising: (a) contacting the MGLL polypeptide with said ligand in a suitable solution comprising PEG MME 5K, Na Citrate pH 5.5, and n-Octyl-Beta-D-Glucopyranoside; and, b) crystallizing said resulting complex of MGLL polypeptide-ligand from said solution. In one embodiment, the MGLL polypeptide is a polypeptide SEQ ID NO: 7. In another embodiment, PEG MME has an average molecular weight range from 2000 to 10000, wherein said PEG MME is present in solution at a range from about 1% w/v to about 5% w/v and said n-Octyl-Beta-D-Glucopyranoside is present in solution at a range of from about 0.2% to 2%. In a preferred embodiment, PEG MME has an average molecular weight of about 5000 and is present in solution at about 2.4% w/v and said n-Octyl-Beta-D-Glucopyranoside is present in solution at about 0.6%.
The invention further includes a method for the production of a crystal comprising MGLL and a ligand wherein the ligand is a small molecule inhibitor comprising crystallizing a peptide comprising SEQ ID NO: 7 with a potential inhibitor.
The invention includes a method for identifying a potential inhibitor of MGLL comprising: a) using a three dimensional structure of MGLL as defined by atomic coordinates according to Table 5; b) replacing one or more MGLL amino acids selected from SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 in said three-dimensional structure with a different amino acid to produce a modified MGLL; c) using said three-dimensional structure to design or select said potential inhibitor; d) synthesizing said potential inhibitor; and, e) contacting said potential inhibitor with said modified MGLL in the presence of a substrate to test the ability of said potential inhibitor to inhibit MGLL or said modified MGLL. In another embodiment, the potential inhibitor is selected from a database. In a preferred embodiment, the potential inhibitor is designed de novo. In another preferred embodiment, the potential inhibitor is designed from a known inhibitor. In a highly preferred embodiment, the step of employing said three-dimensional structure to design or select said potential inhibitor comprises the steps of: a) identifying chemical entities or fragments capable of associating with modified MGLL; and b) assembling the identified chemical entities or fragments into a single molecule to provide the structure of said potential inhibitor. In one embodiment, the potential inhibitor is a competitive inhibitor of SEQ ID NO: 7. In a different embodiment, the potential inhibitor is a non-competitive or uncompetitive inhibitor of SEQ ID NO: 7. In yet another embodiment, an inhibitor is identified by the method.
Engineered Forms and Fragments
Engineered forms of MGLL or fragments thereof, for instance engineered forms or fragments comprising active sites defined by two or more amino acids selected from the group consisting of: SER48-HIS54, ARG57, TYR58, HIS77, HIS80, MET88, PHE93, PHE96, GLY120-ILE127, ILE145-VAL161, ALA163, SER176-ASN195, ASP197, ILE200, CYS201, ALA203, LEU205-VAL207, PHE209-SER218, ASP239-ASP243, TYR268-LEU275, THR279 may be prepared by any available means including synthetic or recombinant means. Such fragments may then be used in the assays as described herein, for example, but not limited to, high-throughput assays to detect interactions between prospective agents and the active site within the fragment.
For recombinant expression or production of the forms or fragments of the invention, nucleic acid molecules encoding the form or fragment may be prepared. Nucleic acid molecules encoding engineered forms or fragments of the invention may differ in sequence because of the degeneracy in the genetic code or may differ in sequence as they encode proteins or protein fragments that differ in amino acid sequence. Homology or sequence identity between two or more such nucleic acid molecules is determined by BLAST (Basic Local Alignment Search Tool) analysis using the algorithm employed by the programs blastp, blastn, blastx, tblastn and tblastx (Karlin and Altschul 1990) and (Altschul 1993), fully incorporated by reference, which are tailored for sequence similarity searching.
The approach used by the BLAST program is to first consider similar segments between a query sequence and a database sequence, then to evaluate the statistical significance of all matches that are identified and finally to summarize only those matches which satisfy a preselected threshold of significance. For a discussion of basic issues in similarity searching of sequence databases, see (Altschul et al. 1994) which is fully incorporated by reference. The search parameters for histogram, descriptions, alignments, expect (i.e., the statistical significance threshold for reporting matches against database sequences), cutoff, matrix and filter are at the default settings. For a discussion of default scoring matrix used by blastp, blastx, tblastn, and tblastx, see (Henikoff 1992).
The encoding nucleic acid molecules of the present invention or fragments thereof (i.e., synthetic oligonucleotides) and those that are used as probes or specific primers for polymerase chain reaction (PCR) or to synthesize gene sequences encoding proteins of the invention can easily be synthesized by chemical techniques, for example, the phosphotriester method of (Matteucci and Caruthers 1981) or using automated synthesis methods. In addition, larger DNA segments can readily be prepared by well-known methods, such as synthesis of a group of oligonucleotides that define various modular segments of the gene, followed by ligation of oligonucleotides to build the complete modified gene.
The encoding nucleic acid molecules of the present invention may further be modified so as to contain a detectable label for diagnostic and probe purposes. A variety of such labels are known in the art and can readily be employed with the encoding molecules herein described. Suitable labels include, but are not limited to, biotin, radiolabeled nucleotides and the like. A skilled artisan can employ any of the art-known labels to obtain a labeled encoding nucleic acid molecule.
The present invention further provides recombinant DNA molecules (rDNA) that contain a coding sequence for a protein or protein fragment as described herein. As used herein, an rDNA molecule is a DNA molecule that has been subjected to molecular manipulation. Methods for generating rDNA molecules are well known in the art, for example, see (Sambrook et al. 1989). In the preferred rDNA molecules, a coding DNA sequence is operably linked to expression control sequences and/or vector sequences.
The choice of vector and expression control sequences to which one of the protein encoding sequences of the present invention is operably linked depends directly, as is well known in the art, on the functional properties desired (e.g., protein expression, and the host cell to be transformed). A vector of the present invention may be capable of directing the replication or insertion into the host chromosome, and preferably also expression, of the structural gene included in the rDNA molecule.
Expression control elements that are used for regulating the expression of an operably linked protein encoding sequence are known in the art and include, but are not limited to, inducible promoters, constitutive promoters, secretion signals, and other regulatory elements. Preferably, the inducible promoter is readily controlled, such as being responsive to a nutrient in the host cell's medium.
The present invention further provides host cells transformed with a nucleic acid molecule that encodes a protein or protein fragment of the present invention. The host cell can be either prokaryotic or eukaryotic. Eukaryotic cells useful for expression of a protein of the invention are not limited, so long as the cell line is compatible with cell culture methods and compatible with the propagation of the expression vector and expression of the gene product. Preferred eukaryotic host cells include, but are not limited to, insect, yeast, and mammalian cells. Preferred eukaryotic host cells include Spodoptera frugiperda (Sf9 or Sf21) insect cells.
Transformed host cells of the invention may be cultured under conditions that allow the production of the recombinant protein. Optionally the recombinant protein is isolated from the medium or from the cells; recovery and purification of the protein may not be necessary in some instances where some impurities may be tolerated.
Kits may also be prepared with any of the above described nucleic acid molecules, proteins, protein fragments, vector and/or host cells optionally packaged with the reagents needed for a specific assay, such as those described above. In such kits, the protein, protein fragments, or other reagents may be attached to a solid support, such as glass or plastic beads.
High-Throughput Assays
Compound identification methods can be performed using conventional laboratory assay formats or in high-throughput assays, including, but not limited to, those described below.
Immunoassays are a group of techniques used for the measurement of specific biochemical substances, commonly at low concentrations in complex mixtures such as biological fluids. The assays depend upon suitably prepared and selected antibodies with specificity and high affinity for their complementary antigens. A substance to be measured must, of necessity, be antigenic, either an immunogenic macromolecule or a haptenic small molecule. To each sample a known limited amount of specific antibody is added and the fraction of the antigen combining with it, often expressed as the bound:free ratio, is estimated by quantifying the signal from the antibody. Quantification can be achieved with a number of readily identifiable labels and used for various types of assays, including, but not limited to, radioisotopes for radioimmunoassays (RIA), fluorescent molecules for fluoroimmunoassays (FIA), stable free radicals for spin immunoassays, chemiluminescent molecules for chemiluminescent immunoassays (CLIA), colloidal gold particles for immunogold assays, and enzymes for enzyme-linked immunosorbent assays (ELISA).
A common immunoassay format is the ELISA, which avoids the hazards of radiochemicals and the expense of fluorescence detection systems. Instead, an ELISA is a form of quantitative immunoassay based on the use of antibodies (or antigens) that may be linked to an insoluble carrier surface, which is then used to “capture” the relevant antigen (or antibody) the test solution. The antigen-antibody complex is then detected by measuring the activity of an appropriate enzyme that can be covalently attached to the capture antigen (or antibody) or to a subsequent “detection” antibody (or antigen). For more information on ELISA techniques, see, for example, (Crowther 1995); (Kemeny (editor) and Challacombe (editor) 1988), (Kemeny 1991), and (Ishikawa 1999).
Colorimetric assays for enzymes are methods of quantitative chemical analysis in which the concentration or amount of a compound is determined by comparing the color produced by the reaction of a reagent with both standard and test amounts of the compound, often using a calorimeter. A calorimeter is a device for measuring color intensity or differences in color intensity, either visually or photoelectrically. Standard calorimetric assays of beta-galactosidase enzymatic activity are well known to those skilled in the art, see for example, (Norton and Coffin 1985). A calorimetric assay can be performed on whole cell lysates using O-nitrophenyl-beta-D-galacto-pyranoside (ONPG, Sigma) as the substrate in a standard calorimetric beta-galactosidase assay (Sambrook et al. 1989). Automated calorimetric assays are also available for the detection of beta-galactosidase activity, as described in U.S. Pat. No. 5,733,720.
Enzymatic substrates that become fluorescent after being acted upon by an enzyme generally are well known. Such fluorescent substrates typically have two components that are bound to one another through, for example, a covalent chemical bond. One component is a fluorescent molecule that is capable of fluorescing by first accepting light energy and then emitting light energy. The other component is an entity that prevents the fluorescent molecule from accepting or emitting light energy when the two components are covalently bound to one another. In the presence of an appropriate enzyme, the enzyme cleaves the covalent bond between the two components and separates one component from the other to permit the fluorescent molecule to accept and emit light energy. In other words, the enzyme frees the fluorescent molecule and allows it to fluoresce. Ideally, fluorescent substrates should be soluble and stable in aqueous buffers, should have a high affinity for the enzymes that act upon them, and should yield a strong signal upon enzymatic action (U.S. Pat. No. 5,998,593A).
Detecting fluorescence emitted from the fluorescent component of a fluorescent enzyme substrate is typically achieved in two steps. In particular, the fluorescent molecule is first excited with light energy and subsequently the fluorescence emitted from the fluorescent component is then detected. Generally, fluorescent molecules can be excited with light energy from, for example, a laser or another suitable light source. Fluorescence is detected with a device designed to detect light energy of a wavelength that is emitted by the fluorescent molecule. Such excitation and emission detection systems generally are designed to operate at particular wavelength ranges (U.S. Pat. No. 5,998,593A).
Thermofluor® assays detect small changes in the intrinsic melting temperature of proteins based on binding of ligands. Compounds that interact preferentially with the native form of the protein will increase the Tm, the temperature at which half of the protein is unfolded (Pantoliano et al. 2001). The technique monitors changes in the fluorescent intensity of dyes such as 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS). The fluorescent dyes are quenched in aqueous environments but increase in fluorescence on binding to the hydrophobic core of denatured proteins.
Modeling the Three-Dimensional Structure of MGLL
The atomic coordinate data provided in Table 5, or the coordinate data derived from homologous proteins may be used to build a three-dimensional model of MGLL. Any available computational methods may be used to build the three dimensional model. As a starting point, the X-ray diffraction pattern obtained from the assemblage of the molecules or atoms in a crystalline version of MGLL or an MGLL homolog can be used to build an electron density map using tools well known to those skilled in the art of crystallography and X-ray diffraction techniques. Additional phase information extracted either from the diffraction data and available in the published literature and/or from supplementing experiments may then be used to complete the reconstruction.
For basic concepts and procedures of collecting, analyzing, and utilizing X-ray diffraction data for the construction of electron densities see, for example, (Campbell 1984), (Cantor and Schimmel 1980), (Brunger 1993), (Woolfson 1997), (Drenth 1999), (Tsirelson and Ozerov 1996), and (U.S. Pat. No. 5,942,428A); (U.S. Pat. No. 6,037,117A); (U.S. Pat. No. 5,200,910A); and (U.S. Pat. No. 5,365,456A), each of which is herein specifically incorporated by reference in their entirety.
For basic information on molecular modeling, see, for example, (Schlecht 1998); (Gans et al. 1996); (Cohen (editor) 1996); and (Smith 1996). U.S. patents which provide detailed information on molecular modeling include (U.S. Pat. No. 4,906,122A; U.S. Pat. No. 5,030,103A; U.S. Pat. No. 5,583,973A; U.S. Pat. No. 5,612,894A; U.S. Pat. No. 5,994,503A; U.S. Pat. No. 6,071,700A; U.S. Pat. No. 6,075,014A; U.S. Pat. No. 6,075,123A; U.S. Pat. No. 6,080,576A; U.S. Pat. No. 6,093,573A), each of which are incorporated by reference herein in their entirety.
Methods of Using the Atomic Coordinates to Identify and Design Ligands of Interest
The atomic coordinates of the invention, such as those described in Table 5, or coordinates substantially identical to or homologous to those of Table 5 may be used with any available methods to prepare three dimensional models of MGLL as well as to identify and design MGLL ligands, inhibitors or antagonists or agonist molecules. Such a method provides the amino acid sequence and/or X-ray diffraction data in a form which allows a skilled artisan to analyze and molecular model the three-dimensional structure of MGLL or related molecules, including a subdomain thereof.
For instance, three-dimensional modeling may be performed using the experimentally determined coordinates derived from X-ray diffraction patterns, such as those in Table 5, for example, wherein such modeling includes, but is not limited to, drawing pictures of the actual structures, building physical models of the actual structures, and determining the structures of related subunits and MGLL/ligand and MGLL subunit/ligand complexes using the coordinates. Such molecular modeling can utilize known X-ray diffraction molecular modeling algorithms or molecular modeling software to generate atomic coordinates corresponding to the three-dimensional structure of MGLL.
As described above, molecular modeling involves the use of computational methods, preferably computer assisted methods, to build realistic models of molecules that are identifiably related in sequence to the known crystal structure. It also involves modeling new small molecule inhibitors bound to MGLL starting with the structures of MGLL and or MGLL complexed with known ligands or inhibitors. The methods utilized in ligand modeling range from molecular graphics (i.e., 3D representations) to computational chemistry (i.e., calculations of the physical and chemical properties) to make predictions about the binding of ligands or activities of ligands; to design new ligands; and to predict novel molecules, including ligands such as drugs, for chemical synthesis, collectively referred to as rational drug design.
One approach to rational drug design is to search for known molecular structures that might bind to an active site. Using molecular modeling, rational drug design programs can look at a range of different molecular structures of drugs that may fit into the active site of an enzyme, and by moving them in a three-dimensional environment it can be decided which structures actually fit the site well.
An alternative but related rational drug design approach starts with the known structure of a complex with a small molecule ligand and models modifications of that small molecule in an effort to make additional favourable interactions with MGLL.
The present invention includes the use of molecular and computer modeling techniques to design and select and design ligands, such as small molecule agonists or antagonists or other therapeutic agents that interact with MGLL. For example, the invention as herein described includes the design of ligands that act as competitive inhibitors of at least one MGLL function by binding to all, or a portion of, the active sites or other regions of MGLL.
This invention also includes the design of compounds that act as uncompetitive inhibitors of at least one function of MGLL. These inhibitors may bind to all, or a portion of, the active sites or other regions of MGLL already bound to its substrate and may be more potent and less non-specific than competitive inhibitors that compete for MGLL active sites. Similarly, non-competitive inhibitors that bind to and inhibit at least one function of MGLL whether or not it is bound to another chemical entity may be designed using the atomic coordinates of MGLL or complexes comprising MGLL of this invention.
The atomic coordinates of the present invention also provide the needed information to probe a crystal of MGLL with molecules composed of a variety of different chemical features to determine optimal sites for interaction between candidate inhibitors and/or activators and MGLL. For example, high resolution X-ray diffraction data collected from crystals saturated with solvent allows the determination of where each type of solvent molecule sticks. Small molecules that bind to those sites can then be designed and synthesized and tested for their inhibitory activity (Travis 1993).
The present invention also includes methods for computationally screening small molecule databases and libraries for chemical entities, agents, ligands, or compounds that can bind in whole, or in part, to MGLL. In this screening, the quality of fit of such entities or compounds to the binding site or sites may be judged either by shape complementarity or by estimated interaction energy (Meng et al. 1992).
The design of compounds that bind to, promote or inhibit the functional activity of MGLL according to this invention generally involves consideration of two factors. First, the compound must be capable of physically and structurally associating with MGLL. Non-covalent molecular interactions important in the association of MGLL with the compound include hydrogen bonding, van der Waals and hydrophobic interactions. Second, the compound must be able to assume a conformation that allows it to associate with MGLL. Although certain portions of the compound may not directly participate in the association with MGLL, those portions may still influence the overall conformation of the molecule. This, in turn, may have a significant impact on binding affinities, therapeutic efficacy, drug-like qualities and potency. Such conformational requirements include the overall three-dimensional structure and orientation of the chemical entity or compound in relation to all or a portion of the active site or other region of MGLL, or the spacing between functional groups of a compound comprising several chemical entities that directly interact with MGLL.
The potential, predicted, inhibitory agonist, antagonist or binding effect of a ligand or other compound on MGLL 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 MGLL, synthesis and testing of the compound may be obviated. However, if computer modeling indicates a strong interaction, the molecule may then be synthesized and tested for its ability to interact with MGLL. In this manner, synthesis of inoperative compounds may be avoided. In some cases, inactive compounds are synthesized predicted on modeling and then tested to develop a SAR (structure-activity relationship) for compounds interacting with a specific region of MGLL.
One skilled in the art may use one of several methods to screen chemical entities fragments, compounds, or agents for their ability to associate with MGLL and more particularly with the individual binding pockets or active sites of MGLL. This process may begin by visual inspection of, for example, the active site on the computer screen based on the atomic coordinates of MGLL or MGLL complexed with a ligand. Selected chemical entities, compounds, or agents may then be positioned in a variety of orientations, or docked within an individual binding pocket of MGLL. Docking may be accomplished using software such as QUANTA, available from Accelrys, Inc., San Diego, Calif.; and SYBYL, available for Tripos, St. Louis, Mo.; followed by energy minimization and molecular dynamics with standard molecular mechanics forcefields, such as CHARMm; available from Accelrys, Inc., San Diego, Calif.; and AMBER, University of California, San Francisco.
Specialized computer programs may also assist in the process of selecting chemical entities. These include but are not limited to: GRID (Goodford 1985), available from Oxford University, Oxford, UK); MCSS (Miranker and Karplus 1991), available from Molecular Simulations, Burlington, Mass.; AUTODOCK (Goodsell and Olsen 1990), available from Scripps Research Institute, La Jolla, Calif.; and DOCK (Kuntz et al. 1982), available from University of California, San Francisco, Calif.
The use of software such as GRID, a program that determines probable interaction sites between probes with various functional group characteristics and the macromolecular surface, is used to analyze the surface sites to determine structures of similar inhibiting proteins or compounds. The GRID calculations, with suitable inhibiting groups on molecules (e.g., protonated primary amines) as the probe, are used to identify potential hotspots around accessible positions at suitable energy contour levels. The program DOCK may be used to analyze an active site or ligand-binding site and suggest ligands with complementary steric properties.
Once suitable chemical entities, compounds, or agents have been selected, they can be assembled into a single ligand or compound or inhibitor or activator. Assembly may proceed by visual inspection of the relationship of the fragments to each other on the three-dimensional image. This may be followed by manual model building using software such as QUANTA or SYBYL.
Useful programs to aid in connecting the individual chemical entities, compounds, or agents include but are not limited to: CAVEAT (Bartlett et al. 1989); 3D Database systems such as MACCS-3D (Martin 1992), available from MDL Information Systems, San Leandro, Calif.; and HOOK, available from Molecular Simulations, Burlington, Mass.
Several methodologies for searching three-dimensional databases to test pharmacophore hypotheses and select compounds for screening are available. These include the program CAVEAT (Bacon and Moult 1992). For instance, CAVEAT uses databases of cyclic compounds which can act as “spacers” to connect any number of chemical fragments already positioned in the active site. This allows one skilled in the art to quickly generate hundreds of possible ways to connect the fragments already known or suspected to be necessary for tight binding.
Instead of proceeding to build an inhibitor activator, agonist or antagonist of MGLL in a step-wise fashion one chemical entity at a time as described above, such compounds may be designed as a whole or “de novo” using either an empty active site or optionally including some portion(s) of a known molecule(s). These methods include: LUDI (Bohm 1992), available from Biosym Technologies, San Diego, Calif.; LEGEND (Nishibata and Itai 1991), available from Molecular Simulations, Burlington, Mass.; and LeapFrog, available from Tripos Associates, St. Louis, Mo., USA.
For instance, the program LUDI can determine a list of interaction sites into which to place both hydrogen bonding and hydrophobic fragments. LUDI then uses a library of linkers to connect up to four different interaction sites into fragments. Then smaller “bridging” groups such as —CH2- and —COO— are used to connect these fragments. For example, for the enzyme DHFR, the placements of key functional groups in the well-known inhibitor methotrexate were reproduced by LUDI. See also, (Rotstein and Murcko 1993).
Other molecular modeling techniques may also be employed in accordance with this invention. See, e.g., (Cohen et al. 1990). See also, (Navia and Murcko 1992).
Once a compound has been designed or selected by the above methods, the affinity with which that compound may bind or associate with MGLL may be tested and optimized by computational evaluation and/or by testing biological activity after synthesizing the compound. Inhibitors or compounds may interact with the MGLL 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 compound binds to MGLL.
A compound designed or selected as binding or associating with MGLL may be further computationally optimized so that in its bound state it would preferably lack repulsive electrostatic interaction with MGLL. Such non-complementary (e.g., electrostatic) interactions include repulsive charge-charge, dipole-dipole and charge-dipole interactions. Specifically, the sum of all electrostatic interactions between the inhibitor and MGLL when the inhibitor is bound, preferably make a neutral or favourable contribution to the enthalpy of binding. Weak binding compounds will also be designed by these methods so as to determine SAR.
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 et al. 1992); AMBER, University of California, San Francisco; QUANTA and CHARMm, available from Accelrys, Inc., San Diego, Calif.; and Insight II/Discover, from Biosysm Technologies Inc., San Diego, Calif., USA. Other hardware systems and software packages will be known to those skilled in the art.
Once a compound that associates with MGLL has been optimally selected or designed, as described above, substitutions may then be made in some of its atoms or side 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. It should, of course, be understood that components known in the art to alter conformation may be avoided. Such substituted chemical compounds may then be analyzed for efficiency of fit to MGLL by the same computer methods described in detail, above.
Use of Homology Structure Modeling to Design Ligands with Modulated Binding or Activity to MGLL.
The present invention includes the use of the atomic coordinates and structures of MGLL and/or MGLL complexed with an inhibitor to design modifications to starting compounds and derivatives thereof that will bind more tightly or interact more specifically to the target enzyme.
The structure of a complex between the MGLL and the starting compound can be used to guide the modification of that compound to produce new compounds that have other desirable properties for applicable industrial and other uses (e.g., as pharmaceuticals), such as chemical stability, solubility or membrane permeability. (Lipinski et al. 1997).
Binding compounds, agonists, antagonists and such that are known in the art. Such compounds can be diffused into or soaked with the stabilized crystals of MGLL to form a complex for collecting X-ray diffraction data. Alternatively, the compounds, known and unknown in the art, can be cocrystallized with MGLL by mixing the compound with MGLL before precipitation.
To produce custom high affinity and very specific compounds, the structure of MGLL can be compared to the structure of a selected non-targeted molecule and a hybrid constructed by changing the structure of residues at the binding site for a ligand for the residues at the same positions of the non-target molecule. The process whereby this modeling is achieved is referred to as homology structure modeling. This is done computationally by removing the side chains from the molecule or target of known structure and replacing them with the side chains of the unknown structure put in sterically plausible positions. In this way it can be understood how the shapes of the active site cavities of the targeted and non-targeted molecules differ. This process, therefore, provides information concerning how a bound ligand can be chemically altered in order to produce compounds that will bind tightly and specifically to the desired target but will simultaneously be sterically prevented from binding to the non-targeted molecule. Likewise, knowledge of portions of the bound ligands that are facing to the solvent would allow introduction of other functional groups for additional pharmaceutical purposes. The use of homology structure modeling to design molecules (ligands) that bind more tightly to the target enzyme than to the non-target enzyme has wide spread applicability.
Databases and Computer Systems
An amino acid sequence or nucleotide sequence of MGLL and/or X-ray diffraction data, useful for computer molecular modeling of MGLL or a portion thereof, can be provided in a variety of mediums to facilitate use thereof. In one application of this embodiment, databases comprising data pertaining to MGLL, or at least one subdomain thereof, amino acid and nucleic acid sequence and/or X-ray diffraction data of the present invention is recorded on computer readable medium. A skilled artisan can readily appreciate how any of the presently known computer readable media can be used to create a manufacture comprising computer readable medium having recorded thereon an amino acid sequence and/or X-ray diffraction data of the present invention.
A variety of data storage structures are available to a skilled artisan for creating a computer readable medium having recorded thereon an amino acid sequence and/or atomic coordinate/X-ray diffraction data of the present invention. 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 of the present invention on computer readable media. The sequence information can be represented in a word processing text file, formatted in commercially-available software such as WordPerfect and MICROSOFT Word, or represented in the form of an ASCII file, stored in a database application, such as DB2, Sybase, Oracle, or the like. A skilled artisan can readily adapt any number of dataprocessor structuring formats (e.g., text file or database) in order to obtain computer readable media having recorded thereon the information of the present invention.
By providing computer readable media having sequence and/or atomic coordinates based on X-ray diffraction data, a skilled artisan can routinely access the sequence and atomic coordinate or X-ray diffraction data to model a related molecule, a subdomain, mimetic, or a ligand thereof. Computer algorithms are publicly and commercially available which allow a skilled artisan to access this data provided in a computer readable medium and analyze it for molecular modeling and/or RDD (rational drug design). See, e.g., (Mary Ann Liebert (Publishers) 1995).
The present invention further provides systems, particularly computer-based systems, which contain the sequence and/or diffraction data described herein. Such systems are designed to do structure determination and RDD for MGLL or at least one subdomain thereof. Non-limiting examples are microcomputer workstations available from Silicon Graphics Incorporated and Sun Microsystems running UNIX based, Windows NT or IBM OS/2 operating systems.
A variety of comparing means can also be used to compare a target sequence or target motif with the data storage means to identify structural motifs or electron density maps derived in part from the atomic coordinate/X-ray diffraction data. A skilled artisan can readily recognize that any one of the publicly available computer modeling programs can be used as the search means for the computer-based systems of the present invention.
Integrated Procedures Which Utilize the Present Invention
Molecular modeling is provided by the present invention for rational drug design (RDD) of mimetics and ligands of MGLL. As described above, the drug design paradigm uses computer-modeling programs to determine potential mimetics and ligands which are expected to interact with sites on the protein. The potential mimetics or ligands are then screened for activity and/or binding and/or interaction. For MGLL-related mimetics or ligands, screening methods can be selected from assays for at least one biological activity of MGLL, e.g., such as hydrolysis by MGLL.
Thus, the tools and methodologies provided by the present invention may be used in procedures for identifying and designing ligands which bind in desirable ways with the target. Such procedures utilize an iterative process whereby ligands are synthesized, tested and characterized. New ligands can be designed based on the information gained in the testing and characterization of the initial ligands and then such newly identified ligands can themselves be tested and characterized. This series of processes may be repeated as many times as necessary to obtain ligands with the desirable binding properties.
The following steps (1-7) serve as an example of the overall procedure:

    
    
        1. A biological activity of a target is selected (e.g., hydrolysis by MGLL).
        2. A ligand is identified that appears to be in some way associated with the chosen biological activity (e.g., the ligand may be an inhibitor of a known activity). The activity of the ligand may be tested by in vivo and/or in vitro methods. A ligand of the present invention can be, but is not limited to, at least one selected from a lipid, a nucleic acid, a compound, a protein, an element, an antibody, a saccharide, an isotope, a carbohydrate, an imaging agent, a lipoprotein, a glycoprotein, an enzyme, a detectable probe, and antibody or fragment thereof, or any combination thereof, which can be detectably labeled as for labeling antibodies. Such labels include, but are not limited to, enzymatic labels, radioisotope or radioactive compounds or elements, fluorescent compounds or metals, chemiluminescent compounds and bioluminescent compounds. Alternatively, any other known diagnostic or therapeutic agent can be used in a method of the invention. Suitable compounds are then tested for activities in relationship to the target. Complexes between MGLL and ligands are made either by co-crystallization or more commonly by diffusing the small molecule ligand into the crystal. X-ray diffraction data from the complex crystal are measured and a difference electron density map is calculated. This process provides the precise location of the bound ligand on the target molecule. The difference Fourier is calculated using measure diffraction amplitudes and the phases of these reflections calculated from the coordinates.
        3. Using the methods of the present invention, X-ray crystallography is utilized to create electron density maps and/or molecular models of the interaction of the ligand with the target molecule. The entry of the coordinates of the target into the computer programs discussed above results in the calculation of most probable structure of the macromolecule. These structures are combined and refined by additional calculations using such programs to determine the probable or actual three-dimensional structure of the target including potential or actual active or binding sites of ligands. Such molecular modeling (and related) programs useful for rational drug design of ligands or mimetics are also provided by the present invention.
        4. The electron density maps and/or molecular models obtained in Step 3 are compared to the electron density maps and/or molecular models of a non-ligand containing target and the observed/calculated differences are used to specifically locate the binding of the ligand on the target or subunit.
        5. Modeling tools, such as computational chemistry and computer modeling, are used to adjust or modify the structure of the ligand so that it can make additional or different interactions with the target. The ligand design uses computer-modeling programs which calculate how different molecules interact with the various sites of the target, subunit, or a fragment thereof. Thus, this procedure determines potential ligands or ligand mimetics.
        6. The newly designed ligand from Step 5 can be tested for its biological activity using appropriate in vivo or in vitro tests, including the high-throughput screening methods discussed above. The potential ligands or mimetics are then screened for activity relating to MGLL, or at least a fragment thereof. Such screening methods are selected from assays for at least one biological activity of the native target. The resulting ligands or mimetics, provided by methods of the present invention, are useful for treating, screening or preventing diseases in animals, such as mammals (including humans).
        7. Of course, each of the above steps can be modified as desired by those of skill in the art so as to refine the procedure for the particular goal in mind. Also, additional X-ray diffraction data may be collected on MGLL, MGLL/ligand complexes, MGLL structural target motifs and MGLL subunit/ligand complexes at any step or phase of the procedure. Such additional diffraction data can be used to reconstruct electron density maps and molecular models, which may further assist in the design and selection of ligands with the desirable binding attributes.
    
    


It is to be understood that the present invention is considered to include stereoisomers as well as optical isomers, e.g., mixtures of enantiomers as well as individual enantiomers and diastereomers, which arise as a consequence of structural asymmetry in selected compounds, ligands or mimetics of the present series.
Some of the compounds or agents disclosed or discovered by the methods herein may contain one or more asymmetric centers and thus give rise to enantiomers, diastereomers, and other stereoisomeric forms. The present invention is also meant to encompass all such possible forms as well as their racemic and resolved forms and mixtures thereof. When the compounds described or discovered herein contain olefinic double bonds or other centers of geometric asymmetry, and unless otherwise specified, it is intended to include both E and Z geometric isomers. All tautomers are intended to be encompassed by the present invention as well.
EXAMPLES
Without further description, it is believed that one of ordinary skill in the art can, using the preceding description and the following illustrative examples, make and utilize the present invention and practice the claimed methods. The following working examples therefore, specifically point out preferred embodiments of the present invention, and are not to be construed as limiting in any way the remainder of the disclosure.
BLAST Search and Sequence Alignment
The reference sequences for MGLL Isoform 2 (Karlsson et al. 2001) and Isoform 1 (Wall et al. 1997) are shown in FIG. 1: A and FIG. 1: B, respectively. Note that the sequences of MGLL Isoform 2 and MGLL Isoform 1 are 100% identical, except that MGLL Isoform 1 has an additional 10 amino acids at the N-terminus. The alignment for MGLL Isoform 2 and MGLL Isoform 1 is shown in FIG. 1: C. The numbering of amino acids for MGLL Isoform 2 (SEQ ID NO: 1) was used throughout the following description to refer to amino acids in the engineered constructs of the present invention.
A BLAST search (Altschul et al. 1990) against the sequence data set deposited in the Protein Data Bank (Sussman et al. 1998) was conducted to identify a protein of known crystal structure with reasonable sequence homology to MGLL. The closest relative to MGLL found in the Protein Data Bank was RsbQ, a stress-response regulator in Bacillus subtilis (Kaneko et al. 2005). RsbQ shares 25% sequence identity with human MGLL. RsbQ also had the highest sequence identity and smallest insertions and deletions relative to MGLL of any structure available in the PDB at the time of this work. The protein RsbQ, PDB ID 1wom (Kaneko et al. 2005), was used as a template. The sequences of RsbQ and MGLL isoform 2 were aligned using the ClustalW software (Thompson et al. 1994; Higgins et al. 1996). RsbQ is a α/β hydrolase with a catalytic triad composed of Ser96, His247 and Asp219. The Asp-His-Ser catalytic triad of MGLL matched the corresponding residues in RsbQ. This alignment was duplicated within the GeneMine software (Lee and Irizarry 2001) and adjusted to eliminate insertions or deletions within elements of secondary structure without disturbing the alignment of the catalytic residues. The final alignment of human MGLL isoform 2 and RsbQ is shown in FIG. 2A. Although the sequence identity between MGLL and RsbQ is low for generating a homology model for MGLL, it was estimated that a low accuracy model would be sufficient to identify hydrophobic residues in the outside of the molecule that could trigger aggregation.
Homology Model
A homology model of MGLL was created using RsbQ as a template and the “quick refine” option in GeneMine software (FIG. 2A) (Levitt 1992) (Lee and Irizarry 2001). The model shows a α/β hydrolase domain and a cap-domain composed of 4 helices. Helix 151-185 of the cap-domain shows amphiphilic properties, characteristic of proteins involved in lipid binding. The helix contains a slight bend due to the presence of a proline residue at position 172. 14 out of the 32 residues constituting the helix are hydrophobic (FIG. 2A). 7 of the hydrophobic residues are Leucines (Leu 152, 157, 159, 171, 174, 176, 184). In the model, the side chains of Leu 159 and 176 point towards the solvent, possibly constituting a recognition site for the interaction of MGLL with the membrane. It should be noted that because of the low accuracy of the model, other Leucine residues, whose side chains appear to point toward the core of the molecule in the homology model, may also contribute to the hydrophobic properties of MGLL and trigger the need for detergent.
Construct Design
A library of constructs was designed in an effort to generate MGLL protein that would be less prone to aggregation, not require detergent for purification, and be more suitable for high-throughput screening and crystallization. A total of 52 mut-MGLL constructs were generated by mixing and matching the cap-domain mutations, surface mutations, and truncations (Table 1). Seven different hydrophobic Leucine residues (designated as Leu or L) of the cap-domain were selected for mutations (Leu 162, 167, 169, 171, 174, 176, and 184). The Leucine residues were replaced by Serine (designated as Ser or S), Glutamine (designated as Gln, or Q), or Arginine (designated as Arg or R). In addition, eight Lysine residues (designated as Lys or K) were identified at the surface of the MGLL homology model (Lys 36, 160, 165, 188, 206, 226, 259 and 269) and were mutated to Alanine (designated as Ala or A) to increase crystal contacts, promote crystallization, and improve crystal quality. The surface mutations were introduced into the mut-MGLL (hMGLL 1-303 L169S, 176S) double cap-domain mutant construct either independently or in combination with other surface mutations. N-terminal and C-terminal truncation constructs were also designed (Table 1). The N-terminus was truncated at amino acid 9, 19, 26, and 33. The C-terminus was truncated at 297 and 292. The N-terminal and C-terminal truncations were introduced independently or combined with other truncations and were introduced into the mut-MGLL (hMGLL 1-303 L169S, 176S) double cap-domain mutant construct (Table 1). All constructs, including the wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) construct, were engineered with an N-terminal histidine tag (His tag) followed by a TEV protease cleavage site so that the tag could be cleaved after purification. TEV is highly site-specific protease that is found in the Tobacco Etch Virus (Invitrogen).
Cloning
The cDNA for MGLL was cloned from human brain DNA and used as a template to generate a PCR fragment of full-length wt-MGLL corresponding to amino acids 1-303 of the reference sequence for human MGLL Isoform 2 (SEQ ID NO: 1) The sequences for the 3′ and 5′ PCR primers are shown below.









5′ primer:



gagaatttggtattttcaaggtatgccagaggaaagttcccc



(SEQ ID NO: 8)


 



3′ primer:



tggatgtgtatgtttctatcagggtggggacgaagttcc



(SEQ ID NO: 9)






The PCR product was purified (GENECLEAN SPIN kits, Qbiogene, Inc), treated with T4 polymerase (New England Biolabs), ligated into the modified pENTR.11cLIC vector, and transformed into TOP10 one shot competent cells (Invitrogen). After sequence confirmation, the mutations were added by Quickchange mutagenesis, (Stratagene). The sequence confirmed plasmids were purified for transfection into insect cells using the BaculoDirect Baculovirus Expression System (Invitrogen). All of the resulting proteins contained an N-terminal His tag followed by a TEV cleavage site and the amino acids of the different MGLL constructs. Viral stock was propagated for two more amplifications at a low multiplicity of infection (MOI) to render a P2 virus stock.
Recombinant Production of wt-MGLL and mut-MGLL
Large-scale expression was carried out in 2-liter shake flasks or WAVE bioreactors (WAVE Products Group, GE Healthcare). The P2 virus was expanded to generate a high titer P3 stock by infecting Sf9 cells in suspension at MOI of 0.3 and harvesting the virus after 72 hours. Cell paste for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and mut-MGLL were obtained by infecting Sf9 cells at a density of 1.5×106 cells/ml with a MOI of 1. Infected cultures were maintained at 27° C. under constant shaking at 140 rpm. Cells were harvested 65-72 hours post-infection by centrifugation at 1000×g for 10 minutes at 4° C. Cell viability were determined by Guava ViaCount or Trypan Blue and routinely were between 60 and 80% at time of harvest. Cell pellets were washed once in phosphate-buffered saline with broad range protease inhibitors and stored at −80° C.
Purification of Wild-Type MGLL (wt-MGLL)
A pilot purification of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3), performed in the complete absence of detergent, generated no protein (data not shown). A second purification of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) was done with detergent in the lysis buffer only. Frozen cell pellets for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) were thawed, resuspended, and lysed in Bugbuster® lysis buffer for 1 hour at 4° C. Bugbuster® lysis buffer is a proprietary lysis buffer from Invitrogen that contains detergent. The lysate was clarified by centrifugation at 40,000×g for 1 hr. No detergent was added at this point or during the rest of the purification. From this point forward, the purification protocol and buffers were the same as described below for mut-MGLL. An average of 2.2 mg of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) per liter of cell culture was obtained. Further analysis by size exclusion chromatography showed complete aggregation of the purified wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3), which confirmed the need for detergent for wt-MGLL purification as previously described in the literature.
Purification of mut-MGLL
Mutant MGLL (mut-MGLL) constructs were purified in the absence of detergent. Frozen cell pellets were thawed and resuspended in buffer A (50 mM Hepes buffer pH 7.5, 400 mM NaCl, 5% glycerol, 0.05% BME, 1× Complete EDTA-free protease inhibitor cocktail tablets (Roche)), dounce homogenized and mechanically lysed with a microfluidizer processor (Microfluidics). The extract was clarified by centrifugation at 40,000×g for 1 hr. The cleared lysate was loaded on a 1 ml His-Trap FF Crude column (GE-Healthcare) at 4° C. using the AktaXpress system. For larger preparations, a 5 ml His-Trap FF Crude column was used. The column was washed with 10-15 column volumes (CV) of buffer A containing 30 mM imidazole and mut-MGLL was eluted with 5 CV of 50 mM Hepes buffer pH 7.5, 400 mM NaCl, 5% glycerol, 0.05% BME, 400 mM imidazole. In most preparations, 30 mM imidazole was included in Buffer A from the beginning of the preparation to reduce non-specific binding on the His-Trap column. In addition, a slightly lower imidazole concentration of 350 mM was used in the final elution during later preparations to further improve purity. Glycerol concentration was also reduced to 4% to avoid back pressure problems on the AktaXpress, after it was determined that mut-MGLL constructs were stable in 2% glycerol. The elution peak was directly loaded on a Superdex 200 HR 16/60 preequilibrated with 50 mM Hepes pH7.5 buffer containing 200 mM NaCl, 2% glycerol, 2 mM DTT, 2 mM EDTA.DTT. Fractions were analysed by SDS-PAGE. Fractions containing mut-MGLL were pooled. Purification yields were determined by Bradford assay using the protein assay kit from BioRad according to manufacturer's instruction with BSA as a standard (Bradford 1976).
The majority of constructs containing N-terminal and/or C-terminal truncations did not have high enough expression to allow for purification of soluble protein (Table 1).
Constructs that were evaluated containing just the cap-domain mutations generated between 0.7 and 4.5 mg/L, except the mut-MGLL construct containing the L174Q mutation, which showed no expression (Table 1). Analysis by size exclusion chromatography showed that the purified mut-MGLL proteins were 90% monomeric and only 10% aggregated compared to 100% aggregation for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) (FIG. 1A), which indicated that the mutations significantly improved protein solubility and eliminated the need for detergent during purification.
Constructs that were evaluated with a combination of cap-domain and surface mutations showed expression levels between 0.5 and 3.6 mg/L and were also  ̃90% monomeric on SDS Page as well (data not shown).
TEV Cleavage
To remove the N-terminal His tag, 0.2 units of TEV Protease for each ug of mut-MGLL were added to the mut-MGLL pool. The reaction was done overnight at 4° C. Cleavage of the histidine tag was monitored by SDS-PAGE.
Complex Formation
For crystallization trials, compounds were added in a 1:2 molar ratio (mut-MGLL:compound). TEV cleaved mut-MGLL was first diluted to 0.3 mg/ml with buffer containing 50 mM Hepes pH 7.5, 200 mM NaCl, 2% Glycerol, 2 mM DTT, and 2 mM EDTA. Compounds were added to the diluted protein and the mixture was incubated overnight at 4° C. After the overnight incubation, the mixture was concentrated to a final protein concentration of 6.0 mg/ml using a Ultrafree membrane (10 KDa cut-off). At this stage the purity was >98% as determined by SDS-PAGE and the protein was ready for crystallization trials.
Circular Dichroism (CD)
One construct, TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5), was selected for further characterization by CD to ensure that the mutations introduced did not adversely affect protein conformation and activity. Circular dichroism experiments were performed on a Circular Dichroism Spectrometer Model 202 from Aviv Instruments Inc. The CD scans of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5) (5 μM protein in 10 mM cacodylic acid pH 7 and 140 mM NaCl) were measured from 200 to 260 nm. Temperature melts were monitored at 210 nm. The CD spectra were converted to molar ellipticity and are shown in FIG. 1B.
The CD scans for the wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) purified in the presence of detergent and TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5) were similar indicating that the two enzymes had a similar conformation (FIG. 1B). The scans were characteristic of proteins with high alpha-helical content as expected for a lipase.
Kinetic Analysis
To ensure that the mutations engineered did not adversely affect protein activity, a number of the newly generated MGLL mutants were analyzed using by enzyme assay and then compared to wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3). A small fluorescent substrate, 4-methyl coumarin butyrate (4MC-B) was used to compare the activity of the engineered mutants to the activity of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3). The catalytic efficiency (kcat/KM) for the hydrolysis of the 4MC-B was similar for wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and all MGLL mutants tested (Table 2). A larger more aliphatic fluorescent substrate, Coumarin Arachidonate (C-A), structurally more closely related to the MGLL natural substrate, 2-AG, was used to compare the activity of wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) to TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5). The catalytic efficiency for the hydrolysis of the C-A substrate was equivalent between wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5), which confirmed that the mutations did not affect MGLL activity (Table 2).
The Michaelis-Menten parameters for the hydrolysis of 4-methylcoumarin butyrate (4MC-B) and coumarin arachidonate (C-A) substrate were determined using 4-5 nM of MGLL in 20 mM Pipes pH 7 and 150 mM NaCl at 37° C. The change in fluorescence due to substrate hydrolysis was monitored using excitation/emission wavelengths of 335/440 in a Safire II instrument from Tecan. The hyperbolic rates versus substrate concentration curves for the hydrolysis of 4MC-B were fit to the Michaelis-Menten equation using Excel.




  v
  =
  
    
      
        V
        max
      
      *
      
        [
        S
        ]
      
    
    
      
        K
        M
      
      +
      
        [
        S
        ]
      
    
  




The solubility limit of coumarin arachidonate (C-A) substrate did not allow for the determination of KM and kcat. The apparent kcat/KM ratio for the hydrolysis of C-A was determined at [S]<<KM. The apparent KM for C-A was estimated to be >30 μM. The kcat/KM values reported are the average from independent values determined from five substrate concentrations ranging from 700 to 40 nM.
Thermal Stability
The Thermofluor® assay is a powerful tool to screen for small molecule inhibitors interacting with a protein's active site or allosteric site. The assay detects small changes in the intrinsic melting temperature of proteins based on binding of ligands. Compounds that interact preferentially with the native form of the protein will increase the Tm, the temperature at which half of the protein is unfolded (Pantoliano et al. 2001). The technique monitors changes in the fluorescent intensity of dyes such as 1-anilinonaphthalene-8-sulfonic acid (1,8-ANS). The fluorescent dyes are quenched in aqueous environments but increase in fluorescence on binding to the hydrophobic core of denatured proteins.
Thermofluor® assays were conducted to characterize wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) and mut-MGLL (hMGLL 1-303 L169S, L176S) and evaluate if the MGLL mutants could be used for high-throughput screening using Thermofluor®. Three microliters of protein at a concentration of 0.05 mg/ml in 50 mM Pipes pH 7, 200 mM NaCl, 100 μM 1,8-ANS, and 0.001% Tween was added to pre-dispensed compound plates. Wells were overlaid with silicone oil (1 μL, Fluka, type DC 200) to prevent evaporation. Final compound concentrations varied from 150 to 0.15 μM. Assay plates were heated at a rate of 1° C./min for all experiments over a temperature range sufficient to measure protein unfolding. Fluorescence was measured by continuous illumination with UV light (Hamamatsu LC6) supplied via fiber optic and filtered through a custom band-pass filter (380-400 nm; >60D cutoff). Fluorescence emission was detected by measuring light intensity using a CCD camera (Sensys, Roper Scientific) filtered to detect emission at 500±25 nm, resulting in simultaneous and independent readings of all 384 wells. One or more images were collected at each temperature, and the sum of the pixel intensity in a given area of the assay plate was recorded vs temperature, and fit to standard equations to yield the Tm.
The study using Thermofluor® showed that wt-MGLL (hMGLL 1-303) (SEQ ID NO: 3) had a very poor transition, characteristic for aggregated or unfolded proteins (FIG. 5). TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S) (SEQ ID NO: 5), however, gave a strong transition with a Tm value of 56.7° C., indicating that the engineered mutation produced a more soluble MGLL protein that was suitable for high-throughput screening.
Crystallization
All mutants were purified according to the procedure described above and submitted for crystallization trials. Purity greater than 95% as determine by SDS Page was achieved for all constructs. Combinations of high-throughput and manual crystallization screens were used. Several constructs generated crystals but only crystallization conditions containing detergent yielded crystals (data not shown). Apo proteins generated crystals diffracting between 8.0 Å and 9.0 Å only, despite extensive optimization trials. Co-crystallization with methyl arachidonyl fluorophosphonate (MAFP) did not significantly improve diffraction.
Co-crystallization of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) with Compound 1 generated crystals that diffracted to 2.3 Å, but with diffused scattering in one orientation. Further optimization experiments did not improve data quality with that complex. High quality diffraction was achieved by co-crystallization of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) with Compound 2, a compound that was 10 fold more potent than Compound 1. Crystallization of the TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) and Compound 2 complex was achieved at 22° C. with a hanging droplet containing 6 mg/ml protein solution combined with a modified well solution containing 8% polyethylene glycol monomethyl ether 5000 molecular weight (PEG MME 5K), 100 mM Na Citrate pH 5.5 and 2% n-Octyl-Beta-D-Glucopyranoside (OBG), which was suspended over a well solution containing 6% PEG MME 5K, 100 mM Na Citrate pH 5.5 and 2% OBG. Crystals, however, were not generated spontaneously. A seed solution generated from poor quality crystals obtained previously was used to seed crystallization droplets. The optimal volume ratios for obtaining good quality crystals were 1 ul protein solution, 0.5 ul well solution, and 0.2 ul diluted seed stock solution. Any increase in protein concentration resulted in heavy showers and stacked plate crystals despite adjustments in crystallization reagent concentration. Protein supplied at concentrations higher than 6 mg/ml and diluted to 6 mg/ml before crystallization trials also resulted in heavy crystal showers. Final resolution was 1.3 Å for the TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) and Compound 2 complex.
Structure Determination
Crystals were harvested, transferred to 16% PEG MME 5K, 100 mM Na-MES pH 6.0, 25% glycerol and flash frozen in liquid nitrogen. Datasets were collected on a Rigaku M007HF generator at 100K or at the ID19 beamline at IMCA-CAD at the Advanced Photon Source, Chicago. A summary of the data-collection statistics is in Table 3. The data was processed in the HKL2000 suite (Otwinowski and Minor 1997) and the structure was solved by molecular replacement using a modified structure of “Non-haem bromoperoxidase BPO-A1” (PDB ID 1A8Q) as search model in PHASER (McCoy et al. 2007). The initial rebuilding was performed using the default protocol in the AutoBuild Wizard in PHENIX (Adams et al. 2002; Adams et al. 2004; Terwilliger et al. 2008), refinement and automated water picking was carried out in PHENIX.refine (Adams et al. 2002; Adams et al. 2004; Terwilliger et al. 2008); Coot (Emsley and Cowtan 2004) was employed for model building, ligand placement and manual assignment of water molecules. Ligand restraints were generated in PHENIX.elbow (Adams et al. 2002; Adams et al. 2004) and the final model validated using tools implemented in Coot; Figures were generated in PyMol (DeLano 2002). Coordinates for the structure of the complex of TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) with Compound 2 are included as Table 5.
Overall Structure of MGLL
MGLL is part of the sub-family of lipid hydrolases, which in turn is part of a larger family of α/β-hydrolases with diverse catalytic functions. Members of this super-family include: ester hydrolases, lipid hydrolases, thioester hydrolases, peptide hydrolases, haloperoxidases, dehalogenases epoxide hydrolases and C—C bond breaking enzymes (Holmquist 2000). All of these enzymes share a common folding motif called the α/β-hydrolase fold (Ollis et al. 1992; Heikinheimo et al. 1999). This fold is characterized by eight β-sheets flanked on both sides by α-helices. β-sheet 2 is antiparallel to the other sheets and the first and last helix (α1 or αA and α6 or αF) are located on one side of the sheets, whereas the remainder of the helices are present on the opposite side. The α/β-hydrolase fold tolerates a vide variety of inserts without losing the core folding motif. These inserts serve to modify and regulate the catalytic activity of the respective proteins. They can occur in several locations, but are mostly located in a loop region between strand β6 and helix α6.
Herein is described the structure of the inhibitor bound form of human MGLL Isoform 2 (TEV cleaved mut-MGLL (hMGLL 1-303 L169S, L176S, K36A) (SEQ ID NO: 7) with Compound 2), which has been determined by molecular replacement to a resolution of 1.3 Å. The structure of MGLL conforms very closely to the canonical α/β-hydrolase fold. The structure is characterized by eight β-sheets, which form a partial β-barrel adorned on both sides with eight α-helices. MGLL contains two additional helices (α4 (αD′1) and α5 D′2)), which are part of the cap-domain and are inserted in the protein sequence between sheet β6 and helix α6 (αD). Helices α1 (αA) and (α8 (αF) are located on the concave side of the barrel and helices α2 (αB), α3 (αC), α6(D) and α7(E) are on the convex side. Both cap-domain helices are oriented in front of the molecule perpendicular to the plane of the β-barrel.
Interestingly the overall structure of this mammalian MGLL is closer to bacterial lipases than any mammalian lipase when the structure is compared to the latest release of the Protein Databank using the protein structure-matching tool (SSM) at the European Institute of Bioinformatics (EBI) (Boutselakis et al. 2003). The 3D-alignment produces several close hits against bacterial Bromoperoxidases, Chloroperoxidases and Arylesterases. The same hits were also produced by a PHI-blast search of the Protein Databank against the protein sequence alone. Since no similar 3D-hits were found against any diacylglycerol lipases or triacylglycerol lipases, it can be inferred that the structural requirements for cleavage of triacylglycerol and diacylglycerol esters are substantially different from those required for the cleavage of mono-glycerol esters. It appears as if these classes of proteins, even though they perform similar functions, represent a different branch on the evolutionary tree of lipases.
Superposition of several hits from the 3D-alignment (Chloroperoxidase L, PDBID:1A88; Bromoperoxidase A1, PDBID:1A8Q; P. putida Esterase, PDBID:1ZOI; Gamma Lactamase, PDBID: 1HKH) shows that the (α/β-hydroxylase core without the cap-domain superimposes very well (Table 4 and FIG. 10). The largest differences are seen in the first 20 residues of the N-terminus of MGLL, for which there is no complement in the other proteins. Further differences are evident in helix α6 (D). In MGLL α6 continuously spans approximately 20 residues, whereas in the other structures it is partially unraveled and split into two pieces connected by a short loop.
MGLL Binding Pocket
Compound 2 is bound in an extended and closed binding pocket, which is located between helices α4, α6, α7 and α5. Even though the solvent accessible surface area of the compound (712 Å2) is fairly large, it is almost completely enclosed by the protein. The protein accomplishes this by employing a so-called “cap-domain”, “lid”, or “flap”, which regulates access to the binding site based on the membrane bound state of the protein. The cap-domain is compromised of residues from helices α4 to α5 (also referred to as αD′1 and αD′2 throughout the literature). The catalytic triad of MGLL consists of residues Ser122, Asp239, His269 and is located in the center of the binding pocket. The catalytic nucleophile Ser122 resides on a tight turn between strand β5 and helix α3, which is also commonly referred to as the “nucleophilic elbow”. The structurally conserved network of hydrogen-bond donors, which comprises the nucleophilic elbow and the loop connecting α1 and β3 (Gly50, Ala51, Met 123 and Gly124) is called the oxyanion hole and serves to stabilize the anionic transition state of the catalytic reaction. The amide carbonyl of Compound 2 points into the oxyanion hole and forms a critical hydrogen bond with the backbone amide nitrogen of Met123 adjacent to the catalytic Serine. The azetidine-piperazine-pyrazine part of the ligand projects into a narrow amphiphilic pocket and fills the available space almost completely. This portion of the ligand does not participate in hydrogen bond interactions with the protein, but one of the pyrazine nitrogens forms an H-bond to a water-network involving two buried water molecules and the side-chains of residues Glu53, Arg57 and His272. A face-to-face π-stacking interaction with of the pyrazine ring with Tyr194 provides further interaction energy.
The binding pocket on the benzoxazole-cyclohexane site of the ligand is less occluded than its counter part on the opposite site. The benzoxazole portion of the ligand is located in a hydrophobic environment constituted mainly from side chains of aliphatic residues. The cyclohexane portion projects into a more spacious void, and along with the benzoxazole, is the only part of the inhibitor, which is accessible by solvent in the protein bound state. These parts of the ligand form mostly van der Waals interactions with the protein. The cyclohexane part of the molecule is less well ordered than the remainder of the ligand. This can be explained by the fact that this region of the cap-domain (α4 and part of the loop connecting to α5) with which the inhibitor interacts, is displays significantly higher temperature factors as compared to the rest of the protein. The elevated temperature factors signify the inherent flexibility of this region, which probably facilitates its displacement from the surface of the protein during ligand binding and release.
Enabling Mutations
In order to obtain the MGLL structure several enabling mutations were required. Two mutations in the lid sub-domain (L169S and L176S) helped to increase solubility of the protein enough to prevent aggregation and to eliminate the need for detergents in protein purification. L169S is located at the C-terminal end of helix α4 (αD′1) and L176S on a loop connecting (α4 to α5. Interestingly, the cap-domain in the engineered protein still contains quite a few surface exposed aliphatic residues, but the mutations are apparently sufficient to reverse the inherent lipophilic character of the protein enough to prevent aggregation in solution.
The K36A surface mutation was inspired by a series of reports indicating that the replacement of flexible residues with high conformational entropy present on the surface of proteins helps to promote crystallization under certain circumstances (Longenecker et al. 2001; Mateja et al. 2002). The K36A mutation is present on a loop connecting sheets β2 and β3. This loop interacts with the cap-domain of a neighboring symmetry related molecule between Val170 and Pro172. Analysis of this packing interaction reveals that the Lysine would have fit snugly into this packing interface, so the immediate reason necessitating its absence for crystallization is not obvious.
The mutation appears nevertheless to be beneficial, since this particular part of the cap-domain exhibits relatively high temperature factors and is less well ordered than other parts of the molecule. It is conceivable that this high dynamic mobility would cause the lid to clash into Lys36 in certain parts of the conformational pool. The K36A mutation would eliminate this potential for clashes and may thus contribute to the successful crystallization of the molecule.
Tables







TABLE 1







Engineered forms of MGLL









Purification



yield mg/L












Cap Mutations



mut-MGLL (1-303) L169S, L176S
4.5


mut-MGLL (1-303) L167Q
2.3


mut-MGLL (1-303) L171Q
0.7


mut-MGLL (1-303) L174Q
0


mut-MGLL (1-303) L167Q, L171Q
5


mut-MGLL (1-303) L167Q, L174Q
7


mut-MGLL (1-303) L171Q, L174Q
1.5


mut-MGLL (1-303) L167Q, L171Q, L174Q
0


mut-MGLL (1-303) L169Q, L176Q
*N.D.


mut-MGLL (1-303) L169S
*N.D.


mut-MGLL (1-303) L176S
*N.D.


mut-MGLL (1-303) L162S
*N.D.


mut-MGLL (1-303) L162Q
*N.D.


mut-MGLL (1-303) L162R
*N.D.


mut-MGLL (1-303) L184S
*N.D.


mut-MGLL (1-303) L184Q
*N.D.


mut-MGLL (1-303) L184R
*N.D.


mut-MGLL (1-303) L169S
*N.D.


mut-MGLL (1-303) L169Q
*N.D.


mut-MGLL (1-303) L169R
*N.D.


mut-MGLL (1-303) L176S
*N.D.


mut-MGLL (1-303) L176Q
*N.D.


mut-MGLL (1-303) L176R
*N.D.


mut-MGLL (1-303) L167S
*N.D.


mut-MGLL (1-303) L167R
*N.D.


mut-MGLL (1-303) L171S
*N.D.


mut-MGLL (1-303) L171R
*N.D.


mut-MGLL (1-303) L174S
*N.D.


mut-MGLL (1-303) L174R
*N.D.


Cap Mutations + Surface Mutations


mut-MGLL (1-303) L169S, L176S, K36A
3.6


mut-MGLL (1-303) L169S, L176S, K160A
0.7


mut-MGLL (1-303) L169S, L176S, K165A
0.5


mut-MGLL (1-303) L169S, L176S, K226A
2.3


mut-MGLL (1-303) L169S, L176S, K36A, K226A
1.5


mut-MGLL (1-303) L169S, L176S, K36A, K188A
*N.D.


mut-MGLL (1-303) L169S, L176S, K36A, K206A
*N.D.


mut-MGLL (1-303) L169S, L176S, K36A, K269A
*N.D.


mut-MGLL (1-303) L169S, L176S, K188A
*N.D.


mut-MGLL (1-303) L169S, L176S, K206A
*N.D.


mut-MGLL (1-303) L169S, L176S, K259A
*N.D.


Cap Mutations + Truncations


mut-MGLL (9-303) L169S, L176S
2


mut-MGLL (9-297) L169S, L176S
1


mut-MGLL (1-292) L169S, L176S
0.5


mut-MGLL (19-303) L169S, L176S
0


mut-MGLL (19-297) L169S, L176S
0


mut-MGLL (19-292) L169S, L176S
0


mut-MGLL (26-303) L169S, L176S
0


mut-MGLL (26-297) L169S, L176S
0


mut-MGLL (26-292) L169S, L176S
0


mut-MGLL (33-303) L169S, L176S
0


mut-MGLL (33-297) L169S, L176S
0


mut-MGLL (33-292) L169S, L176S
0





*N.D. is Not Determined













TABLE 2










 4MC-B


 











KM
Kcat
Kcat/KM


Construct
(uM)
(uM−1min−1)
(uM−1min−1)













wt-MGLL.1-303
162
68
0.42


CAP MUTANTS


mut-MGLL. 1-303, L169S, L176S
136
48
0.35


mut-MGLL 9-303, L169S, L176S
88
27
0.31


mut-MGLL 9-297, L169S, L176S
126
26
0.21


mut-MGLL 1-303, L171Q
105
51
0.48


mut-MGLL 1-303, L167Q, L171Q
84
59
0.71


mut-MGLL. 1-303, L167Q, L174Q
84
70
0.83


mut-MGLL 1-303, L171Q, L174Q
89
47
0.52


CAP + SURFACE MUTANTS


mut-MGLL 1-303, L169S, L176S, K36A
124
51
0.41


mut-MGLL. 1-303, L169S, L176S, K160A
90
30
0.33


mut-MGLL. 1-303, L169S, L176S, K165A
137
27
0.2


mut-MGLL 1-303, L169S, L176S, K226A
110
38
0.35


mut-MGLL 1-303, L169S, L176S, K36A, K226A
123
30
0.25


 





 C-A






Kcat/KM


Construct
(uM−1min−1





wt-MGLL 1-303
0.09


mut-MGLL 1-303, L169S, L176S
0.1


















TABLE 3







mut-MGLL complex



with Compound 2

















Data collection



Wavelength (Å)
1.0


Resolution (Å)
1.3


Space group
C2221


Unit cell parameters (Å)
a = 93.95, b = 128.45, c = 60.6


No. of reflections
336035


No. of unique reflections
79954


Redundancy
4.2 (1.8)


Completeness (%)
89.2 (53.3)


Rmerge
 6.2 (31.9)


I/σ(I)
19.0 (1.75)


Refinement


No. of reflections
75577


No. of reflections in Rfree set
1896


Total No. of non H atoms
2720


No. of protein atoms
2320


No. of ligand atoms
33


No. of solvent molecules
377


R-factor (%)
17.8


Rfree (%)
20.6


R.M.S. Deviation from ideal geometry


Bonds (Å)
0.006


Angles (°)
1.050


B-factors (Å2)


Protein
14.8


Ligand
10.2


Ramachandran Plot


Preferred Regions (%)
96.6


Allowed regions (%)
3.0


Disallowed regions (%)
0.4





















TABLE 4







Chloro-
Bromoperoxidase
P. fluorescens
Gamma



peroxidase L
A1
Arylesterase
Lactamase




















Distance
1.23
1.27
1.29
1.44


[Å]


No. of
168
165
175
173


matching


residues
















TABLE 5





(Table 5 discloses SEQ ID NO: 10):






















CRYST1
93.947
128.145
60.602
90.00
90.00
90.00
C 2 2 21












SCALE1
0.010644
−0.000000
−0.000000
0.00000



SCALE2
0.000000
 0.007804
 0.000000
0.00000



SCALE3
0.000000
 0.000000
 0.016501
0.00000



















ATOM
1
N
PRO
A
7
−24.135
50.110
−0.751
1.00
38.62
N


ATOM
2
CA
PRO
A
7
−24.819
48.812
−0.835
1.00
48.66
C


ATOM
3
C
PRO
A
7
−23.834
47.683
−1.121
1.00
27.13
C


ATOM
4
CB
PRO
A
7
−25.762
48.987
−2.031
1.00
56.71
C


ATOM
5
CG
PRO
A
7
−25.901
50.460
−2.210
1.00
69.43
C


ATOM
6
CD
PRO
A
7
−24.588
51.041
−1.797
1.00
66.62
C


ATOM
7
O
PRO
A
7
−23.163
47.709
−2.152
1.00
27.37
O


ATOM
8
N
ARG
A
8
−23.737
46.710
−0.219
1.00
19.22
N


ATOM
9
CA
ARG
A
8
−22.858
45.567
−0.448
1.00
17.80
C


ATOM
10
C
ARG
A
8
−23.445
44.739
−1.589
1.00
10.71
C


ATOM
11
CB
ARG
A
8
−22.696
44.715
0.818
1.00
16.23
C


ATOM
12
CG
ARG
A
8
−22.085
45.452
2.003
1.00
25.04
C


ATOM
13
CD
ARG
A
8
−20.586
45.608
1.857
1.00
23.77
C


ATOM
14
NE
ARG
A
8
−19.862
44.389
2.217
1.00
19.99
N


ATOM
15
CZ
ARG
A
8
−19.479
44.080
3.453
1.00
18.56
C


ATOM
16
NH1
ARG
A
8
−19.744
44.894
4.472
1.00
16.20
N


ATOM
17
NH2
ARG
A
8
−18.816
42.959
3.671
1.00
13.17
N


ATOM
18
O
ARG
A
8
−24.639
44.451
−1.605
1.00
11.63
O


ATOM
19
N
ARG
A
9
−22.589
44.386
−2.546
1.00
17.31
N


ATOM
20
CA
ARG
A
9
−23.017
43.667
−3.747
1.00
15.23
C


ATOM
21
C
ARG
A
9
−22.202
42.401
−3.960
1.00
12.13
C


ATOM
22
O
ARG
A
9
−21.048
42.328
−3.554
1.00
15.48
O


ATOM
23
CB
ARG
A
9
−22.851
44.558
−4.984
1.00
20.10
C


ATOM
24
CG
ARG
A
9
−23.745
45.788
−5.019
1.00
21.95
C


ATOM
25
CD
ARG
A
9
−23.339
46.711
−6.167
1.00
29.40
C


ATOM
26
NE
ARG
A
9
−24.116
47.949
−6.182
1.00
31.93
N


ATOM
27
CZ
ARG
A
9
−25.269
48.098
−6.824
1.00
41.04
C


ATOM
28
NH1
ARG
A
9
−25.782
47.087
−7.507
1.00
25.69
N


ATOM
29
NH2
ARG
A
9
−25.911
49.259
−6.786
1.00
35.67
N


ATOM
30
N
THR
A
10
−22.815
41.422
−4.621
1.00
13.54
N


ATOM
31
CA
THR
A
10
−22.103
40.220
−5.047
1.00
14.38
C


ATOM
32
C
THR
A
10
−21.025
40.604
−6.058
1.00
21.65
C


ATOM
33
O
THR
A
10
−21.064
41.705
−6.620
1.00
18.22
O


ATOM
34
CB
THR
A
10
−23.056
39.216
−5.709
1.00
14.17
C


ATOM
35
OG1
THR
A
10
−23.504
39.739
−6.972
1.00
14.78
O


ATOM
36
CG2
THR
A
10
−24.260
38.949
−4.824
1.00
13.27
C


ATOM
37
N
PRO
A
11
−20.050
39.708
−6.288
1.00
18.16
N


ATOM
38
CA
PRO
A
11
−19.002
39.977
−7.280
1.00
20.97
C


ATOM
39
C
PRO
A
11
−19.595
40.202
−8.663
1.00
18.43
C


ATOM
40
O
PRO
A
11
−18.911
40.714
−9.553
1.00
28.22
O


ATOM
41
CB
PRO
A
11
−18.166
38.692
−7.261
1.00
23.08
C


ATOM
42
CG
PRO
A
11
−18.376
38.133
−5.880
1.00
20.36
C


ATOM
43
CD
PRO
A
11
−19.814
38.445
−5.561
1.00
18.78
C


ATOM
44
N
GLN
A
12
−20.857
39.834
−8.837
1.00
16.57
N


ATOM
45
CA
GLN
A
12
−21.559
40.039
−10.095
1.00
20.23
C


ATOM
46
C
GLN
A
12
−22.422
41.311
−10.049
1.00
22.29
C


ATOM
47
O
GLN
A
12
−23.222
41.573
−10.953
1.00
24.66
O


ATOM
48
CB
GLN
A
12
−22.396
38.797
−10.439
1.00
25.03
C


ATOM
49
CG
GLN
A
12
−21.571
37.496
−10.571
1.00
15.70
C


ATOM
50
CD
GLN
A
12
−21.138
36.897
−9.229
1.00
17.63
C


ATOM
51
OE1
GLN
A
12
−20.084
36.261
−9.130
1.00
21.07
O


ATOM
52
NE2
GLN
A
12
−21.951
37.096
−8.198
1.00
14.53
N


ATOM
53
N
SER
A
13
−22.250
42.082
−8.974
1.00
21.48
N


ATOM
54
CA
SER
A
13
−22.858
43.409
−8.801
1.00
20.72
C


ATOM
55
C
SER
A
13
−24.332
43.457
−8.369
1.00
23.48
C


ATOM
56
O
SER
A
13
−24.984
44.492
−8.496
1.00
31.32
O


ATOM
57
CB
SER
A
13
−22.605
44.294
−10.028
1.00
28.85
C


ATOM
58
OG
SER
A
13
−21.220
44.583
−10.147
1.00
30.46
O


ATOM
59
N
ILE
A
14
−24.847
42.350
−7.835
1.00
14.51
N


ATOM
60
CA
AILE
A
14
−26.217
42.289
−7.340
0.50
15.83
C


ATOM
61
C
ILE
A
14
−26.235
42.614
−5.846
1.00
16.95
C


ATOM
62
O
ILE
A
14
−25.442
42.063
−5.084
1.00
15.91
O


ATOM
63
CB
AILE
A
14
−26.824
40.881
−7.537
0.50
15.33
C


ATOM
64
CG1
AILE
A
14
−26.665
40.422
−8.991
0.50
17.26
C


ATOM
65
CG2
AILE
A
14
−28.283
40.859
−7.120
0.50
16.38
C


ATOM
66
CD1
AILE
A
14
−27.420
41.277
−9.991
0.50
17.84
C


ATOM
67
CA
BILE
A
14
−26.216
42.309
−7.341
0.50
15.20
C


ATOM
68
CB
BILE
A
14
−26.875
40.935
−7.587
0.50
16.62
C


ATOM
69
CG1
BILE
A
14
−26.944
40.644
−9.088
0.50
33.69
C


ATOM
70
CG2
BILE
A
14
−28.261
40.883
−6.973
0.50
19.08
C


ATOM
71
CD1
BILE
A
14
−27.766
39.427
−9.436
0.50
19.98
C


ATOM
72
N
PRO
A
15
−27.141
43.507
−5.417
1.00
16.97
N


ATOM
73
CA
PRO
A
15
−27.184
43.839
−3.985
1.00
14.74
C


ATOM
74
C
PRO
A
15
−27.555
42.632
−3.125
1.00
12.12
C


ATOM
75
O
PRO
A
15
−28.517
41.925
−3.421
1.00
14.38
O


ATOM
76
CB
PRO
A
15
−28.292
44.898
−3.903
1.00
18.44
C


ATOM
77
CG
PRO
A
15
−28.364
45.482
−5.283
1.00
18.54
C


ATOM
78
CD
PRO
A
15
−28.069
44.335
−6.209
1.00
17.88
C


ATOM
79
N
TYR
A
16
−26.805
42.419
−2.048
1.00
10.96
N


ATOM
80
CA
TYR
A
16
−27.138
41.352
−1.097
1.00
10.67
C


ATOM
81
C
TYR
A
16
−28.503
41.516
−0.436
1.00
12.83
C


ATOM
82
O
TYR
A
16
−29.123
40.536
−0.034
1.00
13.23
O


ATOM
83
CB
TYR
A
16
−26.052
41.183
−0.025
1.00
9.62
C


ATOM
84
CG
TYR
A
16
−24.826
40.476
−0.542
1.00
8.84
C


ATOM
85
CD1
TYR
A
16
−24.876
39.117
−0.866
1.00
8.85
C


ATOM
86
CD2
TYR
A
16
−23.633
41.153
−0.724
1.00
8.95
C


ATOM
87
CE1
TYR
A
16
−23.765
38.465
−1.346
1.00
8.41
C


ATOM
88
CE2
TYR
A
16
−22.515
40.516
−1.210
1.00
10.20
C


ATOM
89
CZ
TYR
A
16
−22.587
39.168
−1.531
1.00
8.74
C


ATOM
90
OH
TYR
A
16
−21.451
38.558
−2.013
1.00
10.69
O


ATOM
91
N
GLN
A
17
−28.983
42.753
−0.314
1.00
14.18
N


ATOM
92
CA
GLN
A
17
−30.280
42.981
0.326
1.00
17.55
C


ATOM
93
C
GLN
A
17
−31.422
42.262
−0.388
1.00
15.26
C


ATOM
94
O
GLN
A
17
−32.478
42.023
0.195
1.00
23.55
O


ATOM
95
CB
GLN
A
17
−30.581
44.479
0.434
1.00
24.16
C


ATOM
96
CG
GLN
A
17
−30.565
45.205
−0.895
1.00
28.39
C


ATOM
97
CD
GLN
A
17
−30.822
46.694
−0.755
1.00
82.31
C


ATOM
98
OE1
GLN
A
17
−31.628
47.121
0.072
1.00
39.90
O


ATOM
99
NE2
GLN
A
17
−30.142
47.492
−1.572
1.00
47.43
N


ATOM
100
N
ASP
A
18
−31.190
41.907
−1.648
1.00
17.60
N


ATOM
101
CA
ASP
A
18
−32.195
41.258
−2.480
1.00
23.44
C


ATOM
102
C
ASP
A
18
−32.059
39.734
−2.474
1.00
19.90
C


ATOM
103
O
ASP
A
18
−32.862
39.029
−3.085
1.00
23.24
O


ATOM
104
CB
ASP
A
18
−32.068
41.765
−3.922
1.00
23.94
C


ATOM
105
CG
ASP
A
18
−32.228
43.271
−4.029
1.00
69.17
C


ATOM
106
OD1
ASP
A
18
−32.980
43.855
−3.221
1.00
35.65
O


ATOM
107
OD2
ASP
A
18
−31.604
43.873
−4.928
1.00
45.97
O


ATOM
108
N
LEU
A
19
−31.041
39.233
−1.785
1.00
14.53
N


ATOM
109
CA
LEU
A
19
−30.671
37.821
−1.885
1.00
16.13
C


ATOM
110
C
LEU
A
19
−30.589
37.165
−0.516
1.00
14.97
C


ATOM
111
O
LEU
A
19
−30.294
37.835
0.469
1.00
13.14
O


ATOM
112
CB
LEU
A
19
−29.303
37.700
−2.550
1.00
14.55
C


ATOM
113
CG
LEU
A
19
−29.136
38.357
−3.924
1.00
17.35
C


ATOM
114
CD1
LEU
A
19
−27.694
38.277
−4.394
1.00
16.13
C


ATOM
115
CD2
LEU
A
19
−30.080
37.710
−4.927
1.00
21.15
C


ATOM
116
N
PRO
A
20
−30.821
35.844
−0.446
1.00
12.60
N


ATOM
117
CA
PRO
A
20
−30.533
35.158
0.818
1.00
9.83
C


ATOM
118
C
PRO
A
20
−29.035
35.204
1.093
1.00
10.92
C


ATOM
119
O
PRO
A
20
−28.221
34.932
0.209
1.00
10.00
O


ATOM
120
CB
PRO
A
20
−30.996
33.714
0.551
1.00
12.85
C


ATOM
121
CG
PRO
A
20
−31.910
33.798
−0.652
1.00
12.45
C


ATOM
122
CD
PRO
A
20
−31.355
34.929
−1.471
1.00
13.91
C


ATOM
123
N
HIS
A
21
−28.659
35.564
2.315
1.00
9.52
N


ATOM
124
CA
HIS
A
21
−27.255
35.688
2.652
1.00
8.77
C


ATOM
125
C
HIS
A
21
−27.013
35.531
4.145
1.00
8.21
C


ATOM
126
O
HIS
A
21
−27.963
35.527
4.940
1.00
11.76
O


ATOM
127
CB
HIS
A
21
−26.707
37.044
2.193
1.00
12.50
C


ATOM
128
CG
HIS
A
21
−27.346
38.212
2.880
1.00
12.08
C


ATOM
129
ND1
HIS
A
21
−28.470
38.836
2.386
1.00
17.03
N


ATOM
130
CD2
HIS
A
21
−27.033
38.852
4.032
1.00
18.83
C


ATOM
131
CE1
HIS
A
21
−28.819
39.819
3.200
1.00
26.65
C


ATOM
132
NE2
HIS
A
21
−27.966
39.847
4.206
1.00
21.10
N


ATOM
133
N
LEU
A
22
−25.747
35.406
4.502
1.00
8.64
N


ATOM
134
CA
LEU
A
22
−25.311
35.446
5.895
1.00
6.76
C


ATOM
135
C
LEU
A
22
−23.982
36.170
5.920
1.00
9.47
C


ATOM
136
O
LEU
A
22
−23.294
36.247
4.898
1.00
14.08
O


ATOM
137
CB
LEU
A
22
−25.178
34.038
6.483
1.00
13.24
C


ATOM
138
CG
LEU
A
22
−24.073
33.100
6.000
1.00
15.59
C


ATOM
139
CD1
LEU
A
22
−22.694
33.547
6.456
1.00
17.18
C


ATOM
140
CD2
LEU
A
22
−24.349
31.688
6.516
1.00
20.78
C


ATOM
141
N
VAL
A
23
−23.620
36.746
7.058
1.00
6.83
N


ATOM
142
CA
VAL
A
23
−22.341
37.409
7.176
1.00
6.32
C


ATOM
143
C
VAL
A
23
−21.464
36.555
8.079
1.00
6.79
C


ATOM
144
O
VAL
A
23
−21.885
36.200
9.196
1.00
7.54
O


ATOM
145
CB
VAL
A
23
−22.500
38.837
7.772
1.00
7.74
C


ATOM
146
CG1
VAL
A
23
−21.163
39.562
7.798
1.00
10.24
C


ATOM
147
CG2
VAL
A
23
−23.538
39.638
6.977
1.00
9.52
C


ATOM
148
N
ASN
A
24
−20.288
36.172
7.603
1.00
6.28
N


ATOM
149
CA
ASN
A
24
−19.422
35.309
8.392
1.00
7.25
C


ATOM
150
C
ASN
A
24
−18.643
36.082
9.467
1.00
8.55
C


ATOM
151
O
ASN
A
24
−18.799
37.308
9.611
1.00
10.11
O


ATOM
152
CB
ASN
A
24
−18.509
34.447
7.496
1.00
6.73
C


ATOM
153
CG
ASN
A
24
−17.357
35.219
6.885
1.00
8.40
C


ATOM
154
OD1
ASN
A
24
−17.116
36.389
7.213
1.00
8.49
O


ATOM
155
ND2
ASN
A
24
−16.601
34.553
6.007
1.00
8.52
N


ATOM
156
N
ALA
A
25
−17.824
35.369
10.233
1.00
8.14
N


ATOM
157
CA
ALA
A
25
−17.137
35.971
11.376
1.00
8.35
C


ATOM
158
C
ALA
A
25
−16.101
37.007
10.966
1.00
11.92
C


ATOM
159
O
ALA
A
25
−15.619
37.777
11.803
1.00
15.43
O


ATOM
160
CB
ALA
A
25
−16.497
34.897
12.238
1.00
12.36
C


ATOM
161
N
ASP
A
26
−15.737
37.019
9.687
1.00
10.81
N


ATOM
162
CA
ASP
A
26
−14.794
37.998
9.162
1.00
11.24
C


ATOM
163
C
ASP
A
26
−15.527
39.170
8.504
1.00
9.54
C


ATOM
164
O
ASP
A
26
−14.896
40.025
7.867
1.00
15.54
O


ATOM
165
CB
ASP
A
26
−13.840
37.328
8.166
1.00
11.66
C


ATOM
166
CG
ASP
A
26
−12.891
36.357
8.840
1.00
22.26
C


ATOM
167
OD1
ASP
A
26
−12.372
36.694
9.930
1.00
24.50
O


ATOM
168
OD2
ASP
A
26
−12.657
35.258
8.288
1.00
19.65
O


ATOM
169
N
GLY
A
27
−16.847
39.201
8.644
1.00
8.36
N


ATOM
170
CA
GLY
A
27
−17.647
40.275
8.091
1.00
10.32
C


ATOM
171
C
GLY
A
27
−17.901
40.175
6.598
1.00
12.61
C


ATOM
172
O
GLY
A
27
−18.324
41.148
5.976
1.00
12.32
O


ATOM
173
N
GLN
A
28
−17.661
38.998
6.021
1.00
9.34
N


ATOM
174
CA
GLN
A
28
−17.881
38.776
4.588
1.00
8.07
C


ATOM
175
C
GLN
A
28
−19.263
38.219
4.330
1.00
6.30
C


ATOM
176
O
GLN
A
28
−19.725
37.332
5.051
1.00
8.36
O


ATOM
177
CB
GLN
A
28
−16.833
37.810
4.037
1.00
9.11
C


ATOM
178
CG
GLN
A
28
−15.413
38.318
4.197
1.00
10.98
C


ATOM
179
CD
GLN
A
28
−14.370
37.258
3.910
1.00
11.09
C


ATOM
180
OE1
GLN
A
28
−13.533
37.414
3.005
1.00
15.32
O


ATOM
181
NE2
GLN
A
28
−14.409
36.177
4.668
1.00
8.14
N


ATOM
182
N
TYR
A
29
−19.950
38.742
3.326
1.00
6.77
N


ATOM
183
CA
TYR
A
29
−21.250
38.230
2.942
1.00
6.19
C


ATOM
184
C
TYR
A
29
−21.135
36.937
2.139
1.00
6.61
C


ATOM
185
O
TYR
A
29
−20.410
36.886
1.130
1.00
8.93
O


ATOM
186
CB
TYR
A
29
−22.002
39.271
2.106
1.00
9.22
C


ATOM
187
CG
TYR
A
29
−22.628
40.389
2.924
1.00
8.73
C


ATOM
188
CD1
TYR
A
29
−21.838
41.288
3.634
1.00
10.30
C


ATOM
189
CD2
TYR
A
29
−23.999
40.536
2.983
1.00
10.59
C


ATOM
190
CE1
TYR
A
29
−22.413
42.307
4.381
1.00
11.19
C


ATOM
191
CE2
TYR
A
29
−24.582
41.561
3.725
1.00
10.78
C


ATOM
192
CZ
TYR
A
29
−23.773
42.435
4.413
1.00
8.82
C


ATOM
193
OH
TYR
A
29
−24.344
43.451
5.158
1.00
12.21
O


ATOM
194
N
LEU
A
30
−21.853
35.911
2.585
1.00
6.17
N


ATOM
195
CA
LEU
A
30
−21.936
34.654
1.848
1.00
5.98
C


ATOM
196
C
LEU
A
30
−23.334
34.514
1.280
1.00
7.23
C


ATOM
197
O
LEU
A
30
−24.333
34.704
1.975
1.00
9.08
O


ATOM
198
CB
LEU
A
30
−21.629
33.462
2.760
1.00
8.48
C


ATOM
199
CG
LEU
A
30
−20.300
33.501
3.519
1.00
8.51
C


ATOM
200
CD1
LEU
A
30
−20.093
32.171
4.263
1.00
11.37
C


ATOM
201
CD2
LEU
A
30
−19.126
33.807
2.603
1.00
10.67
C


ATOM
202
N
PHE
A
31
−23.418
34.168
−0.003
1.00
6.05
N


ATOM
203
CA
PHE
A
31
−24.692
33.900
−0.642
1.00
6.03
C


ATOM
204
C
PHE
A
31
−25.241
32.543
−0.202
1.00
6.46
C


ATOM
205
O
PHE
A
31
−24.484
31.573
−0.167
1.00
6.52
O


ATOM
206
CB
PHE
A
31
−24.508
33.934
−2.167
1.00
6.59
C


ATOM
207
CG
PHE
A
31
−25.711
33.500
−2.921
1.00
7.69
C


ATOM
208
CD1
PHE
A
31
−26.739
34.395
−3.200
1.00
10.06
C


ATOM
209
CD2
PHE
A
31
−25.835
32.185
−3.363
1.00
8.63
C


ATOM
210
CE1
PHE
A
31
−27.869
33.984
−3.895
1.00
12.38
C


ATOM
211
CE2
PHE
A
31
−26.955
31.778
−4.049
1.00
10.13
C


ATOM
212
CZ
PHE
A
31
−27.974
32.674
−4.321
1.00
11.21
C


ATOM
213
N
CYS
A
32
−26.527
32.483
0.110
1.00
5.60
N


ATOM
214
CA
CYS
A
32
−27.173
31.252
0.594
1.00
5.42
C


ATOM
215
C
CYS
A
32
−28.256
30.757
−0.343
1.00
7.15
C


ATOM
216
O
CYS
A
32
−28.868
31.533
−1.082
1.00
7.80
O


ATOM
217
CB
CYS
A
32
−27.782
31.476
1.982
1.00
7.97
C


ATOM
218
SG
CYS
A
32
−26.589
31.940
3.264
1.00
9.76
S


ATOM
219
N
ARG
A
33
−28.524
29.455
−0.263
1.00
6.41
N


ATOM
220
CA
ARG
A
33
−29.521
28.801
−1.095
1.00
6.58
C


ATOM
221
C
ARG
A
33
−30.301
27.808
−0.238
1.00
6.95
C


ATOM
222
O
ARG
A
33
−29.699
27.074
0.576
1.00
9.26
O


ATOM
223
CB
ARG
A
33
−28.790
28.056
−2.217
1.00
12.39
C


ATOM
224
CG
ARG
A
33
−29.458
28.057
−3.570
1.00
21.52
C


ATOM
225
CD
ARG
A
33
−28.543
27.396
−4.608
1.00
13.00
C


ATOM
226
NE
ARG
A
33
−27.652
28.315
−5.317
1.00
9.53
N


ATOM
227
CZ
ARG
A
33
−28.037
29.049
−6.361
1.00
8.94
C


ATOM
228
NH1
ARG
A
33
−29.287
28.978
−6.788
1.00
13.78
N


ATOM
229
NH2
ARG
A
33
−27.172
29.848
−6.972
1.00
10.48
N


ATOM
230
N
TYR
A
34
−31.619
27.773
−0.405
1.00
8.28
N


ATOM
231
CA
TYR
A
34
−32.485
26.905
0.388
1.00
9.23
C


ATOM
232
C
TYR
A
34
−33.474
26.170
−0.498
1.00
12.77
C


ATOM
233
O
TYR
A
34
−33.998
26.735
−1.466
1.00
12.90
O


ATOM
234
CB
TYR
A
34
−33.270
27.722
1.427
1.00
9.73
C


ATOM
235
CG
TYR
A
34
−32.366
28.475
2.366
1.00
10.63
C


ATOM
236
CD1
TYR
A
34
−31.785
27.835
3.453
1.00
11.93
C


ATOM
237
CD2
TYR
A
34
−32.066
29.820
2.157
1.00
10.99
C


ATOM
238
CE1
TYR
A
34
−30.933
28.511
4.301
1.00
12.94
C


ATOM
239
CE2
TYR
A
34
−31.212
30.505
3.005
1.00
9.78
C


ATOM
240
CZ
TYR
A
34
−30.651
29.838
4.072
1.00
10.20
C


ATOM
241
OH
TYR
A
34
−29.795
30.492
4.921
1.00
11.98
O


ATOM
242
N
TRP
A
35
−33.742
24.916
−0.154
1.00
10.36
N


ATOM
243
CA
TRP
A
35
−34.792
24.152
−0.817
1.00
10.15
C


ATOM
244
C
TRP
A
35
−35.623
23.508
0.286
1.00
11.14
C


ATOM
245
O
TRP
A
35
−35.197
22.524
0.909
1.00
11.77
O


ATOM
246
CB
TRP
A
35
−34.206
23.065
−1.721
1.00
10.42
C


ATOM
247
CG
TRP
A
35
−33.285
23.499
−2.831
1.00
12.61
C


ATOM
248
CD1
TRP
A
35
−33.609
23.625
−4.160
1.00
13.17
C


ATOM
249
CD2
TRP
A
35
−31.879
23.777
−2.736
1.00
9.72
C


ATOM
250
NE1
TRP
A
35
−32.500
23.988
−4.884
1.00
13.29
N


ATOM
251
CE2
TRP
A
35
−31.421
24.082
−4.041
1.00
11.08
C


ATOM
252
CE3
TRP
A
35
−30.959
23.791
−1.677
1.00
9.17
C


ATOM
253
CZ2
TRP
A
35
−30.091
24.403
−4.310
1.00
11.75
C


ATOM
254
CZ3
TRP
A
35
−29.643
24.108
−1.943
1.00
9.91
C


ATOM
255
CH2
TRP
A
35
−29.218
24.414
−3.254
1.00
12.71
C


ATOM
256
N
ALA
A
36
−36.793
24.077
0.554
1.00
13.64
N


ATOM
257
CA
ALA
A
36
−37.613
23.643
1.682
1.00
14.33
C


ATOM
258
C
ALA
A
36
−38.871
22.927
1.209
1.00
17.02
C


ATOM
259
CB
ALA
A
36
−37.990
24.850
2.559
1.00
16.53
C


ATOM
260
O
ALA
A
36
−39.523
23.383
0.274
1.00
19.88
O


ATOM
261
N
PRO
A
37
−39.221
21.807
1.859
1.00
14.84
N


ATOM
262
CA
PRO
A
37
−40.460
21.101
1.510
1.00
20.24
C


ATOM
263
C
PRO
A
37
−41.685
21.874
2.015
1.00
18.75
C


ATOM
264
O
PRO
A
37
−41.540
22.811
2.801
1.00
20.85
O


ATOM
265
CB
PRO
A
37
−40.315
19.769
2.257
1.00
19.92
C


ATOM
266
CG
PRO
A
37
−39.457
20.111
3.431
1.00
20.70
C


ATOM
267
CD
PRO
A
37
−38.444
21.075
2.875
1.00
14.82
C


ATOM
268
N
THR
A
38
−42.872
21.479
1.572
1.00
25.39
N


ATOM
269
CA
THR
A
38
−44.096
22.193
1.923
1.00
30.82
C


ATOM
270
C
THR
A
38
−44.415
22.120
3.413
1.00
36.92
C


ATOM
271
O
THR
A
38
−44.878
23.097
4.002
1.00
62.88
O


ATOM
272
CB
THR
A
38
−45.298
21.661
1.127
1.00
40.57
C


ATOM
273
OG1
THR
A
38
−45.516
20.283
1.455
1.00
75.45
O


ATOM
274
CG2
THR
A
38
−45.039
21.786
−0.365
1.00
33.30
C


ATOM
275
N
GLY
A
39
−44.169
20.963
4.019
1.00
26.40
N


ATOM
276
CA
GLY
A
39
−44.489
20.769
5.423
1.00
34.27
C


ATOM
277
C
GLY
A
39
−43.270
20.683
6.320
1.00
44.58
C


ATOM
278
O
GLY
A
39
−42.160
21.033
5.914
1.00
35.00
O


ATOM
279
N
THR
A
40
−43.481
20.224
7.550
1.00
28.10
N


ATOM
280
CA
THR
A
40
−42.392
20.033
8.500
1.00
24.60
C


ATOM
281
C
THR
A
40
−41.346
19.088
7.915
1.00
26.04
C


ATOM
282
O
THR
A
40
−41.672
17.976
7.501
1.00
22.08
O


ATOM
283
CB
THR
A
40
−42.902
19.444
9.829
1.00
30.60
C


ATOM
284
OG1
THR
A
40
−43.947
20.276
10.349
1.00
44.32
O


ATOM
285
CG2
THR
A
40
−41.772
19.352
10.852
1.00
25.32
C


ATOM
286
N
PRO
A
41
−40.083
19.532
7.871
1.00
20.06
N


ATOM
287
CA
PRO
A
41
−39.027
18.659
7.350
1.00
14.83
C


ATOM
288
C
PRO
A
41
−38.722
17.536
8.332
1.00
15.43
C


ATOM
289
O
PRO
A
41
−38.907
17.706
9.542
1.00
18.25
O


ATOM
290
CB
PRO
A
41
−37.807
19.590
7.246
1.00
15.58
C


ATOM
291
CG
PRO
A
41
−38.331
20.986
7.462
1.00
21.18
C


ATOM
292
CD
PRO
A
41
−39.568
20.847
8.283
1.00
20.08
C


ATOM
293
N
LYS
A
42
−38.244
16.407
7.817
1.00
12.49
N


ATOM
294
CA
LYS
A
42
−37.840
15.276
8.648
1.00
15.17
C


ATOM
295
C
LYS
A
42
−36.405
15.425
9.127
1.00
13.51
C


ATOM
296
O
LYS
A
42
−36.002
14.831
10.133
1.00
15.33
O


ATOM
297
CB
LYS
A
42
−37.978
13.968
7.866
1.00
20.90
C


ATOM
298
CG
LYS
A
42
−39.415
13.620
7.520
1.00
33.71
C


ATOM
299
CD
LYS
A
42
−39.587
12.137
7.262
1.00
65.22
C


ATOM
300
CE
LYS
A
42
−38.819
11.702
6.034
1.00
53.35
C


ATOM
301
NZ
LYS
A
42
−39.110
10.284
5.687
1.00
78.02
N


ATOM
302
N
ALA
A
43
−35.630
16.213
8.387
1.00
11.61
N


ATOM
303
CA
ALA
A
43
−34.228
16.414
8.693
1.00
9.88
C


ATOM
304
C
ALA
A
43
−33.707
17.595
7.880
1.00
7.84
C


ATOM
305
O
ALA
A
43
−34.377
18.073
6.961
1.00
9.50
O


ATOM
306
CB
ALA
A
43
−33.421
15.156
8.373
1.00
13.77
C


ATOM
307
N
LEU
A
44
−32.517
18.047
8.250
1.00
8.06
N


ATOM
308
CA
LEU
A
44
−31.815
19.114
7.551
1.00
7.73
C


ATOM
309
C
LEU
A
44
−30.613
18.507
6.858
1.00
7.26
C


ATOM
310
O
LEU
A
44
−30.001
17.577
7.379
1.00
8.67
O


ATOM
311
CB
LEU
A
44
−31.312
20.154
8.560
1.00
10.75
C


ATOM
312
CG
LEU
A
44
−32.320
20.734
9.558
1.00
14.46
C


ATOM
313
CD1
LEU
A
44
−31.600
21.580
10.594
1.00
18.22
C


ATOM
314
CD2
LEU
A
44
−33.367
21.546
8.837
1.00
14.89
C


ATOM
315
N
ILE
A
45
−30.232
19.069
5.718
1.00
7.30
N


ATOM
316
CA
ILE
A
45
−28.991
18.659
5.084
1.00
7.17
C


ATOM
317
C
ILE
A
45
−28.270
19.843
4.445
1.00
6.26
C


ATOM
318
O
ILE
A
45
−28.850
20.592
3.644
1.00
7.44
O


ATOM
319
CB
ILE
A
45
−29.206
17.489
4.079
1.00
7.19
C


ATOM
320
CG1
ILE
A
45
−27.868
16.995
3.511
1.00
7.97
C


ATOM
321
CG2
ILE
A
45
−30.224
17.865
3.008
1.00
9.73
C


ATOM
322
CD1
ILE
A
45
−27.975
15.599
2.857
1.00
11.27
C


ATOM
323
N
PHE
A
46
−27.021
20.011
4.837
1.00
5.88
N


ATOM
324
CA
PHE
A
46
−26.169
21.039
4.251
1.00
5.70
C


ATOM
325
C
PHE
A
46
−25.371
20.481
3.084
1.00
5.52
C


ATOM
326
O
PHE
A
46
−24.738
19.426
3.211
1.00
7.72
O


ATOM
327
CB
PHE
A
46
−25.216
21.629
5.298
1.00
5.92
C


ATOM
328
CG
PHE
A
46
−24.325
22.708
4.757
1.00
6.50
C


ATOM
329
CD1
PHE
A
46
−23.082
22.413
4.216
1.00
7.76
C


ATOM
330
CD2
PHE
A
46
−24.758
24.027
4.769
1.00
7.33
C


ATOM
331
CE1
PHE
A
46
−22.267
23.423
3.693
1.00
7.27
C


ATOM
332
CE2
PHE
A
46
−23.954
25.038
4.250
1.00
8.02
C


ATOM
333
CZ
PHE
A
46
−22.715
24.742
3.711
1.00
8.11
C


ATOM
334
N
VAL
A
47
−25.375
21.211
1.970
1.00
5.46
N


ATOM
335
CA
VAL
A
47
−24.617
20.813
0.783
1.00
6.17
C


ATOM
336
C
VAL
A
47
−23.379
21.693
0.648
1.00
7.61
C


ATOM
337
O
VAL
A
47
−23.491
22.930
0.600
1.00
8.01
O


ATOM
338
CB
VAL
A
47
−25.480
20.904
−0.495
1.00
9.99
C


ATOM
339
CG1
VAL
A
47
−24.642
20.584
−1.753
1.00
10.76
C


ATOM
340
CG2
VAL
A
47
−26.673
19.965
−0.389
1.00
9.71
C


ATOM
341
N
SER
A
48
−22.210
21.053
0.604
1.00
5.98
N


ATOM
342
CA
SER
A
48
−20.915
21.711
0.581
1.00
6.93
C


ATOM
343
C
SER
A
48
−20.251
21.506
−0.791
1.00
7.30
C


ATOM
344
O
SER
A
48
−19.782
20.401
−1.130
1.00
6.87
O


ATOM
345
CB
SER
A
48
−20.035
21.111
1.688
1.00
8.37
C


ATOM
346
OG
SER
A
48
−18.758
21.704
1.745
1.00
8.59
O


ATOM
347
N
HIS
A
49
−20.194
22.579
−1.580
1.00
5.91
N


ATOM
348
CA
HIS
A
49
−19.621
22.526
−2.923
1.00
5.97
C


ATOM
349
C
HIS
A
49
−18.089
22.530
−2.893
1.00
5.83
C


ATOM
350
O
HIS
A
49
−17.461
22.768
−1.861
1.00
5.76
O


ATOM
351
CB
HIS
A
49
−20.161
23.674
−3.800
1.00
6.87
C


ATOM
352
CG
HIS
A
49
−19.608
25.028
−3.457
1.00
4.44
C


ATOM
353
ND1
HIS
A
49
−18.365
25.450
−3.876
1.00
5.38
N


ATOM
354
CD2
HIS
A
49
−20.152
26.072
−2.781
1.00
6.37
C


ATOM
355
CE1
HIS
A
49
−18.151
26.685
−3.445
1.00
7.05
C


ATOM
356
NE2
HIS
A
49
−19.224
27.087
−2.786
1.00
5.73
N


ATOM
357
N
GLY
A
50
−17.475
22.238
−4.039
1.00
6.83
N


ATOM
358
CA
GLY
A
50
−16.033
22.155
−4.121
1.00
6.75
C


ATOM
359
C
GLY
A
50
−15.362
23.413
−4.651
1.00
6.47
C


ATOM
360
O
GLY
A
50
−16.022
24.420
−4.955
1.00
6.59
O


ATOM
361
N
ALA
A
51
−14.050
23.344
−4.781
1.00
5.98
N


ATOM
362
CA
ALA
A
51
−13.268
24.487
−5.244
1.00
5.62
C


ATOM
363
C
ALA
A
51
−13.654
24.862
−6.662
1.00
8.12
C


ATOM
364
O
ALA
A
51
−13.866
23.984
−7.511
1.00
7.71
O


ATOM
365
CB
ALA
A
51
−11.805
24.181
−5.188
1.00
8.37
C


ATOM
366
N
GLY
A
52
−13.719
26.164
−6.927
1.00
6.60
N


ATOM
367
CA
GLY
A
52
−13.966
26.656
−8.270
1.00
9.14
C


ATOM
368
C
GLY
A
52
−15.413
26.649
−8.709
1.00
7.08
C


ATOM
369
O
GLY
A
52
−15.730
27.192
−9.773
1.00
8.55
O


ATOM
370
N
GLU
A
53
−16.302
26.044
−7.927
1.00
5.30
N


ATOM
371
CA
GLU
A
53
−17.719
26.013
−8.276
1.00
6.20
C


ATOM
372
C
GLU
A
53
−18.570
26.767
−7.242
1.00
5.56
C


ATOM
373
O
GLU
A
53
−18.087
27.730
−6.635
1.00
6.90
O


ATOM
374
CB
GLU
A
53
−18.233
24.582
−8.536
1.00
8.29
C


ATOM
375
CG
GLU
A
53
−17.953
23.602
−7.388
1.00
6.83
C


ATOM
376
CD
GLU
A
53
−18.750
22.300
−7.486
1.00
10.34
C


ATOM
377
OE1
GLU
A
53
−19.440
22.076
−8.496
1.00
10.71
O


ATOM
378
OE2
GLU
A
53
−18.712
21.494
−6.522
1.00
9.53
O


ATOM
379
N
HIS
A
54
−19.811
26.356
−7.054
1.00
5.72
N


ATOM
380
CA
HIS
A
54
−20.727
27.085
−6.173
1.00
6.19
C


ATOM
381
C
HIS
A
54
−21.987
26.260
−5.929
1.00
5.74
C


ATOM
382
O
HIS
A
54
−22.179
25.207
−6.563
1.00
7.20
O


ATOM
383
CB
HIS
A
54
−21.089
28.443
−6.801
1.00
7.71
C


ATOM
384
CG
HIS
A
54
−21.857
28.312
−8.079
1.00
6.04
C


ATOM
385
ND1
HIS
A
54
−23.223
28.457
−8.149
1.00
7.69
N


ATOM
386
CD2
HIS
A
54
−21.445
27.990
−9.332
1.00
6.29
C


ATOM
387
CE1
HIS
A
54
−23.622
28.253
−9.395
1.00
8.02
C


ATOM
388
NE2
HIS
A
54
−22.561
27.967
−10.129
1.00
7.68
N


ATOM
389
N
SER
A
55
−22.869
26.749
−5.064
1.00
6.06
N


ATOM
390
CA
SER
A
55
−24.028
25.992
−4.616
1.00
5.79
C


ATOM
391
C
SER
A
55
−25.064
25.729
−5.710
1.00
6.93
C


ATOM
392
O
SER
A
55
−25.864
24.794
−5.609
1.00
8.85
O


ATOM
393
CB
SER
A
55
−24.697
26.711
−3.434
1.00
7.37
C


ATOM
394
OG
SER
A
55
−25.185
27.981
−3.824
1.00
9.12
O


ATOM
395
N
GLY
A
56
−25.093
26.577
−6.736
1.00
7.19
N


ATOM
396
CA
GLY
A
56
−26.086
26.425
−7.789
1.00
9.39
C


ATOM
397
C
GLY
A
56
−25.890
25.189
−8.663
1.00
8.15
C


ATOM
398
O
GLY
A
56
−26.812
24.773
−9.367
1.00
11.61
O


ATOM
399
N
ARG
A
57
−24.706
24.599
−8.591
1.00
7.21
N


ATOM
400
CA
ARG
A
57
−24.409
23.405
−9.371
1.00
8.23
C


ATOM
401
C
ARG
A
57
−25.023
22.147
−8.738
1.00
12.55
C


ATOM
402
O
ARG
A
57
−24.861
21.049
−9.268
1.00
15.41
O


ATOM
403
CB
ARG
A
57
−22.900
23.280
−9.594
1.00
11.50
C


ATOM
404
CG
ARG
A
57
−22.300
24.548
−10.261
1.00
11.08
C


ATOM
405
CD
ARG
A
57
−21.326
24.258
−11.403
1.00
14.23
C


ATOM
406
NE
ARG
A
57
−21.863
23.251
−12.312
1.00
16.18
N


ATOM
407
CZ
ARG
A
57
−22.803
23.478
−13.219
1.00
20.13
C


ATOM
408
NH1
ARG
A
57
−23.305
24.698
−13.381
1.00
23.25
N


ATOM
409
NH2
ARG
A
57
−23.241
22.477
−13.971
1.00
20.33
N


ATOM
410
N
TYR
A
58
−25.759
22.318
−7.639
1.00
8.94
N


ATOM
411
CA
TYR
A
58
−26.351
21.196
−6.889
1.00
8.78
C


ATOM
412
C
TYR
A
58
−27.876
21.187
−6.952
1.00
10.23
C


ATOM
413
O
TYR
A
58
−28.545
20.471
−6.201
1.00
10.07
O


ATOM
414
CB
TYR
A
58
−25.808
21.182
−5.437
1.00
10.33
C


ATOM
415
CG
TYR
A
58
−24.356
20.813
−5.471
1.00
7.86
C


ATOM
416
CD1
TYR
A
58
−23.367
21.764
−5.755
1.00
8.45
C


ATOM
417
CD2
TYR
A
58
−23.968
19.487
−5.324
1.00
9.90
C


ATOM
418
CE1
TYR
A
58
−22.039
21.403
−5.859
1.00
8.11
C


ATOM
419
CE2
TYR
A
58
−22.649
19.117
−5.413
1.00
8.69
C


ATOM
420
CZ
TYR
A
58
−21.682
20.061
−5.696
1.00
9.51
C


ATOM
421
OH
TYR
A
58
−20.374
19.682
−5.811
1.00
12.95
O


ATOM
422
N
GLU
A
59
−28.429
21.975
−7.869
1.00
9.80
N


ATOM
423
CA
GLU
A
59
−29.868
22.136
−7.978
1.00
11.59
C


ATOM
424
C
GLU
A
59
−30.645
20.816
−8.039
1.00
10.87
C


ATOM
425
O
GLU
A
59
−31.608
20.616
−7.297
1.00
10.96
O


ATOM
426
CB
GLU
A
59
−30.203
22.987
−9.208
1.00
13.77
C


ATOM
427
CG
GLU
A
59
−31.654
23.394
−9.288
1.00
14.77
C


ATOM
428
CD
GLU
A
59
−32.069
24.310
−8.144
1.00
16.52
C


ATOM
429
OE1
GLU
A
59
−31.216
25.088
−7.671
1.00
20.67
O


ATOM
430
OE2
GLU
A
59
−33.245
24.251
−7.728
1.00
25.13
O


ATOM
431
N
GLU
A
60
−30.235
19.916
−8.930
1.00
10.53
N


ATOM
432
CA
GLU
A
60
−30.993
18.683
−9.117
1.00
10.68
C


ATOM
433
C
GLU
A
60
−30.896
17.758
−7.908
1.00
9.92
C


ATOM
434
O
GLU
A
60
−31.894
17.181
−7.476
1.00
10.77
O


ATOM
435
CB
GLU
A
60
−30.553
17.959
−10.391
1.00
14.50
C


ATOM
436
CG
GLU
A
60
−30.925
18.708
−11.667
1.00
24.44
C


ATOM
437
CD
GLU
A
60
−32.401
19.063
−11.722
1.00
89.95
C


ATOM
438
OE1
GLU
A
60
−33.241
18.168
−11.485
1.00
55.36
O


ATOM
439
OE2
GLU
A
60
−32.723
20.237
−12.003
1.00
106.69
O


ATOM
440
N
LEU
A
61
−29.694
17.608
−7.372
1.00
8.91
N


ATOM
441
CA
ALEU
A
61
−29.506
16.811
−6.161
0.50
9.95
C


ATOM
442
C
LEU
A
61
−30.339
17.376
−5.013
1.00
10.94
C


ATOM
443
O
LEU
A
61
−31.003
16.637
−4.278
1.00
9.90
O


ATOM
444
CB
ALEU
A
61
−28.031
16.777
−5.757
0.50
11.94
C


ATOM
445
CG
ALEU
A
61
−27.140
15.693
−6.367
0.50
9.56
C


ATOM
446
CD1
ALEU
A
61
−25.700
15.937
−5.975
0.50
8.30
C


ATOM
447
CD2
ALEU
A
61
−27.591
14.303
−5.926
0.50
9.49
C


ATOM
448
CA
BLEU
A
61
−29.513
16.807
−6.168
0.50
9.77
C


ATOM
449
CB
BLEU
A
61
−28.039
16.757
−5.782
0.50
12.63
C


ATOM
450
CG
BLEU
A
61
−27.659
15.622
−4.834
0.50
14.96
C


ATOM
451
CD1
BLEU
A
61
−28.276
14.307
−5.303
0.50
15.85
C


ATOM
452
CD2
BLEU
A
61
−26.151
15.511
−4.742
0.50
14.56
C


ATOM
453
N
ALA
A
62
−30.303
18.694
−4.849
1.00
8.48
N


ATOM
454
CA
ALA
A
62
−31.054
19.331
−3.778
1.00
10.11
C


ATOM
455
C
ALA
A
62
−32.555
19.111
−3.938
1.00
13.13
C


ATOM
456
O
ALA
A
62
−33.267
18.881
−2.955
1.00
10.27
O


ATOM
457
CB
ALA
A
62
−30.732
20.826
−3.722
1.00
10.95
C


ATOM
458
N
ARG
A
63
−33.052
19.194
−5.169
1.00
9.46
N


ATOM
459
CA
ARG
A
63
−34.475
18.973
−5.402
1.00
13.75
C


ATOM
460
C
ARG
A
63
−34.891
17.549
−5.042
1.00
11.63
C


ATOM
461
O
ARG
A
63
−35.964
17.327
−4.481
1.00
12.65
O


ATOM
462
CB
ARG
A
63
−34.846
19.301
−6.849
1.00
11.63
C


ATOM
463
CG
ARG
A
63
−34.838
20.793
−7.131
1.00
15.77
C


ATOM
464
CD
ARG
A
63
−35.202
21.085
−8.579
1.00
27.72
C


ATOM
465
NE
ARG
A
63
−35.167
22.516
−8.871
1.00
33.09
N


ATOM
466
CZ
ARG
A
63
−35.623
23.058
−9.995
1.00
84.41
C


ATOM
467
NH1
ARG
A
63
−36.156
22.291
−10.937
1.00
43.78
N


ATOM
468
NH2
ARG
A
63
−35.549
24.370
−10.175
1.00
46.62
N


ATOM
469
N
MET
A
64
−34.035
16.584
−5.362
1.00
10.52
N


ATOM
470
CA
AMET
A
64
−34.299
15.192
−5.002
0.75
11.44
C


ATOM
471
C
MET
A
64
−34.394
15.041
−3.485
1.00
11.99
C


ATOM
472
O
MET
A
64
−35.315
14.410
−2.965
1.00
12.08
O


ATOM
473
CB
AMET
A
64
−33.212
14.277
−5.575
0.75
11.53
C


ATOM
474
CG
AMET
A
64
−33.399
12.800
−5.229
0.75
11.45
C


ATOM
475
SD
AMET
A
64
−32.651
12.319
−3.663
0.75
14.16
S


ATOM
476
CE
AMET
A
64
−30.923
12.298
−4.124
0.75
13.84
C


ATOM
477
CA
BMET
A
64
−34.318
15.202
−5.003
0.25
12.08
C


ATOM
478
CB
BMET
A
64
−33.259
14.270
−5.583
0.25
12.80
C


ATOM
479
CG
BMET
A
64
−33.680
12.815
−5.583
0.25
26.52
C


ATOM
480
SD
BMET
A
64
−32.270
11.723
−5.382
0.25
12.18
S


ATOM
481
CE
BMET
A
64
−31.712
12.205
−3.742
0.25
0.00
C


ATOM
482
N
LEU
A
65
−33.437
15.628
−2.777
1.00
9.40
N


ATOM
483
CA
LEU
A
65
−33.394
15.534
−1.319
1.00
10.14
C


ATOM
484
C
LEU
A
65
−34.594
16.228
−0.671
1.00
10.86
C


ATOM
485
O
LEU
A
65
−35.174
15.723
0.296
1.00
10.56
O


ATOM
486
CB
LEU
A
65
−32.082
16.117
−0.793
1.00
8.51
C


ATOM
487
CG
LEU
A
65
−30.836
15.282
−1.120
1.00
10.20
C


ATOM
488
CD1
LEU
A
65
−29.562
16.095
−0.943
1.00
11.61
C


ATOM
489
CD2
LEU
A
65
−30.803
14.004
−0.274
1.00
12.05
C


ATOM
490
N
MET
A
66
−34.973
17.383
−1.209
1.00
11.05
N


ATOM
491
CA
AMET
A
66
−36.136
18.088
−0.703
0.50
11.92
C


ATOM
492
C
MET
A
66
−37.395
17.241
−0.931
1.00
14.01
C


ATOM
493
O
MET
A
66
−38.328
17.265
−0.120
1.00
14.48
O


ATOM
494
CB
AMET
A
66
−36.239
19.471
−1.354
0.50
7.66
C


ATOM
495
CG
AMET
A
66
−37.413
20.298
−0.867
0.50
16.42
C


ATOM
496
SD
AMET
A
66
−38.927
19.913
−1.763
0.50
19.91
S


ATOM
497
CE
AMET
A
66
−38.623
20.747
−3.320
0.50
29.87
C


ATOM
498
CA
BMET
A
66
−36.151
18.103
−0.735
0.50
10.75
C


ATOM
499
CB
BMET
A
66
−36.301
19.432
−1.483
0.50
20.22
C


ATOM
500
CG
BMET
A
66
−37.520
20.250
−1.071
0.50
16.81
C


ATOM
501
SD
BMET
A
66
−37.739
21.751
−2.054
0.50
18.63
S


ATOM
502
CE
BMET
A
66
−38.028
21.070
−3.685
0.50
28.73
C


ATOM
503
N
GLY
A
67
−37.404
16.473
−2.021
1.00
12.11
N


ATOM
504
CA
GLY
A
67
−38.501
15.567
−2.320
1.00
18.68
C


ATOM
505
C
GLY
A
67
−38.616
14.423
−1.324
1.00
13.55
C


ATOM
506
O
GLY
A
67
−39.651
13.763
−1.249
1.00
17.32
O


ATOM
507
N
LEU
A
68
−37.550
14.180
−0.570
1.00
14.11
N


ATOM
508
CA
LEU
A
68
−37.555
13.207
0.520
1.00
15.51
C


ATOM
509
C
LEU
A
68
−37.984
13.857
1.838
1.00
15.63
C


ATOM
510
O
LEU
A
68
−37.854
13.257
2.911
1.00
17.55
O


ATOM
511
CB
LEU
A
68
−36.164
12.605
0.693
1.00
14.48
C


ATOM
512
CG
LEU
A
68
−35.579
11.827
−0.482
1.00
16.34
C


ATOM
513
CD1
LEU
A
68
−34.187
11.326
−0.122
1.00
19.31
C


ATOM
514
CD2
LEU
A
68
−36.495
10.672
−0.866
1.00
25.16
C


ATOM
515
N
ASP
A
69
−38.477
15.090
1.742
1.00
15.49
N


ATOM
516
CA
ASP
A
69
−38.936
15.859
2.903
1.00
13.48
C


ATOM
517
C
ASP
A
69
−37.799
16.365
3.800
1.00
14.51
C


ATOM
518
O
ASP
A
69
−37.982
16.554
5.007
1.00
15.76
O


ATOM
519
CB
ASP
A
69
−39.958
15.059
3.724
1.00
18.23
C


ATOM
520
CG
ASP
A
69
−40.801
15.939
4.622
1.00
52.91
C


ATOM
521
OD1
ASP
A
69
−41.032
17.110
4.255
1.00
35.35
O


ATOM
522
OD2
ASP
A
69
−41.236
15.461
5.690
1.00
43.13
O


ATOM
523
N
LEU
A
70
−36.622
16.574
3.219
1.00
11.72
N


ATOM
524
CA
LEU
A
70
−35.529
17.218
3.940
1.00
12.54
C


ATOM
525
C
LEU
A
70
−35.487
18.700
3.571
1.00
11.28
C


ATOM
526
O
LEU
A
70
−35.809
19.083
2.444
1.00
12.36
O


ATOM
527
CB
LEU
A
70
−34.174
16.572
3.621
1.00
12.49
C


ATOM
528
CG
LEU
A
70
−33.854
15.265
4.351
1.00
19.34
C


ATOM
529
CD1
LEU
A
70
−34.549
14.111
3.654
1.00
21.20
C


ATOM
530
CD2
LEU
A
70
−32.350
15.016
4.415
1.00
16.83
C


ATOM
531
N
LEU
A
71
−35.100
19.533
4.530
1.00
9.16
N


ATOM
532
CA
LEU
A
71
−34.777
20.916
4.226
1.00
9.27
C


ATOM
533
C
LEU
A
71
−33.307
20.979
3.834
1.00
8.70
C


ATOM
534
O
LEU
A
71
−32.415
20.689
4.639
1.00
8.84
O


ATOM
535
CB
LEU
A
71
−35.051
21.799
5.440
1.00
10.92
C


ATOM
536
CG
LEU
A
71
−35.014
23.311
5.176
1.00
14.41
C


ATOM
537
CD1
LEU
A
71
−35.732
24.045
6.305
1.00
17.57
C


ATOM
538
CD2
LEU
A
71
−33.592
23.831
5.014
1.00
17.60
C


ATOM
539
N
VAL
A
72
−33.060
21.347
2.586
1.00
7.86
N


ATOM
540
CA
VAL
A
72
−31.708
21.395
2.055
1.00
8.85
C


ATOM
541
C
VAL
A
72
−31.214
22.828
2.109
1.00
8.92
C


ATOM
542
O
VAL
A
72
−31.950
23.753
1.749
1.00
7.86
O


ATOM
543
CB
VAL
A
72
−31.668
20.911
0.591
1.00
7.35
C


ATOM
544
CG1
VAL
A
72
−30.227
20.797
0.097
1.00
9.49
C


ATOM
545
CG2
VAL
A
72
−32.412
19.583
0.451
1.00
9.13
C


ATOM
546
N
PHE
A
73
−29.986
23.021
2.561
1.00
6.35
N


ATOM
547
CA
PHE
A
73
−29.439
24.374
2.612
1.00
6.13
C


ATOM
548
C
PHE
A
73
−27.967
24.386
2.237
1.00
7.10
C


ATOM
549
O
PHE
A
73
−27.275
23.372
2.342
1.00
6.36
O


ATOM
550
CB
PHE
A
73
−29.728
25.081
3.959
1.00
6.94
C


ATOM
551
CG
PHE
A
73
−29.071
24.458
5.160
1.00
7.16
C


ATOM
552
CD1
PHE
A
73
−29.459
23.209
5.624
1.00
8.74
C


ATOM
553
CD2
PHE
A
73
−28.101
25.156
5.873
1.00
9.73
C


ATOM
554
CE1
PHE
A
73
−28.858
22.649
6.746
1.00
11.57
C


ATOM
555
CE2
PHE
A
73
−27.503
24.609
7.010
1.00
11.39
C


ATOM
556
CZ
PHE
A
73
−27.889
23.356
7.450
1.00
11.64
C


ATOM
557
N
ALA
A
74
−27.487
25.535
1.771
1.00
5.58
N


ATOM
558
CA
ALA
A
74
−26.122
25.665
1.312
1.00
6.13
C


ATOM
559
C
ALA
A
74
−25.710
27.124
1.322
1.00
5.53
C


ATOM
560
O
ALA
A
74
−26.562
28.008
1.381
1.00
5.91
O


ATOM
561
CB
ALA
A
74
−25.996
25.112
−0.117
1.00
7.97
C


ATOM
562
N
HIS
A
75
−24.418
27.365
1.229
1.00
5.08
N


ATOM
563
CA
HIS
A
75
−23.942
28.683
0.855
1.00
4.98
C


ATOM
564
C
HIS
A
75
−22.762
28.529
−0.070
1.00
6.30
C


ATOM
565
O
HIS
A
75
−22.151
27.455
−0.160
1.00
6.97
O


ATOM
566
CB
HIS
A
75
−23.603
29.556
2.073
1.00
5.32
C


ATOM
567
CG
HIS
A
75
−22.565
28.985
2.985
1.00
6.21
C


ATOM
568
ND1
HIS
A
75
−22.865
28.561
4.263
1.00
7.36
N


ATOM
569
CD2
HIS
A
75
−21.228
28.821
2.835
1.00
6.89
C


ATOM
570
CE1
HIS
A
75
−21.758
28.146
4.854
1.00
6.18
C


ATOM
571
NE2
HIS
A
75
−20.747
28.289
4.012
1.00
6.86
N


ATOM
572
N
ASP
A
76
−22.428
29.608
−0.771
1.00
5.01
N


ATOM
573
CA
ASP
A
76
−21.200
29.646
−1.548
1.00
5.14
C


ATOM
574
C
ASP
A
76
−20.058
29.931
−0.589
1.00
5.33
C


ATOM
575
O
ASP
A
76
−20.124
30.898
0.189
1.00
5.80
O


ATOM
576
CB
ASP
A
76
−21.280
30.708
−2.658
1.00
6.05
C


ATOM
577
CG
ASP
A
76
−22.301
30.360
−3.726
1.00
7.92
C


ATOM
578
OD1
ASP
A
76
−22.795
29.212
−3.750
1.00
7.77
O


ATOM
579
OD2
ASP
A
76
−22.637
31.243
−4.548
1.00
7.72
O


ATOM
580
N
HIS
A
77
−19.043
29.077
−0.577
1.00
5.59
N


ATOM
581
CA
HIS
A
77
−17.908
29.282
0.298
1.00
5.52
C


ATOM
582
C
HIS
A
77
−17.261
30.624
−0.020
1.00
5.25
C


ATOM
583
O
HIS
A
77
−17.368
31.144
−1.141
1.00
6.90
O


ATOM
584
CB
HIS
A
77
−16.872
28.166
0.122
1.00
7.14
C


ATOM
585
CG
HIS
A
77
−17.404
26.795
0.413
1.00
6.26
C


ATOM
586
ND1
HIS
A
77
−17.922
26.440
1.641
1.00
8.66
N


ATOM
587
CD2
HIS
A
77
−17.465
25.679
−0.357
1.00
7.29
C


ATOM
588
CE1
HIS
A
77
−18.313
25.176
1.605
1.00
9.31
C


ATOM
589
NE2
HIS
A
77
−18.035
24.689
0.407
1.00
9.66
N


ATOM
590
N
VAL
A
78
−16.572
31.196
0.956
1.00
6.80
N


ATOM
591
CA
AVAL
A
78
−15.924
32.466
0.703
0.50
6.73
C


ATOM
592
C
VAL
A
78
−15.026
32.332
−0.531
1.00
7.87
C


ATOM
593
CB
AVAL
A
78
−15.163
32.972
1.947
0.50
15.02
C


ATOM
594
CG1
AVAL
A
78
−13.972
32.076
2.248
0.50
3.02
C


ATOM
595
CG2
AVAL
A
78
−14.756
34.438
1.771
0.50
5.54
C


ATOM
596
O
VAL
A
78
−14.417
31.286
−0.791
1.00
7.09
O


ATOM
597
CA
BVAL
A
78
−15.839
32.437
0.752
0.50
10.12
C


ATOM
598
CB
BVAL
A
78
−14.889
32.695
1.942
0.50
4.36
C


ATOM
599
CG1
BVAL
A
78
−13.801
33.684
1.564
0.50
10.71
C


ATOM
600
CG2
BVAL
A
78
−15.667
33.180
3.151
0.50
15.80
C


ATOM
601
N
GLY
A
79
−15.002
33.392
−1.338
1.00
7.60
N


ATOM
602
CA
GLY
A
79
−14.198
33.399
−2.552
1.00
8.95
C


ATOM
603
C
GLY
A
79
−14.813
32.627
−3.704
1.00
5.87
C


ATOM
604
O
GLY
A
79
−14.131
32.419
−4.721
1.00
8.58
O


ATOM
605
N
HIS
A
80
−16.072
32.226
−3.569
1.00
4.89
N


ATOM
606
CA
HIS
A
80
−16.745
31.427
−4.592
1.00
4.97
C


ATOM
607
C
HIS
A
80
−18.108
31.959
−4.922
1.00
6.40
C


ATOM
608
O
HIS
A
80
−18.766
32.612
−4.100
1.00
6.52
O


ATOM
609
CB
HIS
A
80
−16.888
29.958
−4.138
1.00
5.51
C


ATOM
610
CG
HIS
A
80
−15.581
29.255
−3.999
1.00
7.26
C


ATOM
611
ND1
HIS
A
80
−15.113
28.348
−4.931
1.00
9.83
N


ATOM
612
CD2
HIS
A
80
−14.609
29.374
−3.067
1.00
4.47
C


ATOM
613
CE1
HIS
A
80
−13.918
27.921
−4.561
1.00
4.41
C


ATOM
614
NE2
HIS
A
80
−13.591
28.527
−3.431
1.00
11.14
N


ATOM
615
N
GLY
A
81
−18.558
31.659
−6.134
1.00
7.03
N


ATOM
616
CA
GLY
A
81
−19.903
31.969
−6.550
1.00
7.68
C


ATOM
617
C
GLY
A
81
−20.301
33.409
−6.297
1.00
7.61
C


ATOM
618
O
GLY
A
81
−19.609
34.336
−6.722
1.00
8.19
O


ATOM
619
N
GLN
A
82
−21.410
33.585
−5.595
1.00
7.12
N


ATOM
620
CA
GLN
A
82
−21.947
34.916
−5.350
1.00
7.50
C


ATOM
621
C
GLN
A
82
−21.557
35.462
−3.983
1.00
8.34
C


ATOM
622
O
GLN
A
82
−22.111
36.478
−3.553
1.00
9.53
O


ATOM
623
CB
GLN
A
82
−23.459
34.917
−5.510
1.00
7.66
C


ATOM
624
CG
GLN
A
82
−23.918
34.420
−6.890
1.00
7.96
C


ATOM
625
CD
GLN
A
82
−25.416
34.400
−7.029
1.00
11.20
C


ATOM
626
OE1
GLN
A
82
−26.058
35.446
−7.145
1.00
12.46
O


ATOM
627
NE2
GLN
A
82
−25.998
33.207
−7.017
1.00
9.37
N


ATOM
628
N
SER
A
83
−20.630
34.787
−3.306
1.00
6.24
N


ATOM
629
CA
SER
A
83
−20.130
35.262
−2.022
1.00
5.93
C


ATOM
630
C
SER
A
83
−18.974
36.210
−2.224
1.00
7.64
C


ATOM
631
O
SER
A
83
−18.346
36.241
−3.292
1.00
9.96
O


ATOM
632
CB
SER
A
83
−19.670
34.088
−1.157
1.00
6.90
C


ATOM
633
OG
SER
A
83
−20.781
33.296
−0.820
1.00
5.65
O


ATOM
634
N
GLU
A
84
−18.684
36.989
−1.185
1.00
8.21
N


ATOM
635
CA
GLU
A
84
−17.549
37.900
−1.200
1.00
8.55
C


ATOM
636
C
GLU
A
84
−16.218
37.169
−1.054
1.00
10.87
C


ATOM
637
O
GLU
A
84
−16.181
35.963
−0.796
1.00
10.07
O


ATOM
638
CB
GLU
A
84
−17.701
38.956
−0.097
1.00
8.81
C


ATOM
639
CG
GLU
A
84
−18.819
39.932
−0.371
1.00
10.32
C


ATOM
640
CD
GLU
A
84
−18.897
41.045
0.662
1.00
14.70
C


ATOM
641
OE1
GLU
A
84
−18.584
40.792
1.844
1.00
12.02
O


ATOM
642
OE2
GLU
A
84
−19.285
42.171
0.285
1.00
17.48
O


ATOM
643
N
GLY
A
85
−15.123
37.909
−1.214
1.00
10.41
N


ATOM
644
CA
GLY
A
85
−13.785
37.373
−1.061
1.00
11.16
C


ATOM
645
C
GLY
A
85
−13.025
37.311
−2.371
1.00
12.67
C


ATOM
646
O
GLY
A
85
−13.611
37.083
−3.428
1.00
12.70
O


ATOM
647
N
GLU
A
86
−11.717
37.516
−2.307
1.00
12.86
N


ATOM
648
CA
GLU
A
86
−10.900
37.312
−3.497
1.00
15.49
C


ATOM
649
C
GLU
A
86
−11.139
35.894
−4.012
1.00
14.29
C


ATOM
650
O
GLU
A
86
−11.252
34.955
−3.220
1.00
11.66
O


ATOM
651
CB
GLU
A
86
−9.429
37.536
−3.177
1.00
16.36
C


ATOM
652
CG
GLU
A
86
−9.114
38.976
−2.782
1.00
31.32
C


ATOM
653
CD
GLU
A
86
−7.662
39.174
−2.392
1.00
101.84
C


ATOM
654
OE1
GLU
A
86
−6.862
38.229
−2.567
1.00
54.48
O


ATOM
655
OE2
GLU
A
86
−7.321
40.277
−1.912
1.00
74.42
O


ATOM
656
N
ARG
A
87
−11.240
35.739
−5.329
1.00
12.44
N


ATOM
657
CA
ARG
A
87
−11.629
34.455
−5.910
1.00
11.77
C


ATOM
658
C
ARG
A
87
−10.641
33.329
−5.596
1.00
10.94
C


ATOM
659
O
ARG
A
87
−9.435
33.443
−5.846
1.00
11.69
O


ATOM
660
CB
ARG
A
87
−11.803
34.562
−7.432
1.00
12.45
C


ATOM
661
CG
ARG
A
87
−12.817
35.592
−7.926
1.00
17.92
C


ATOM
662
CD
ARG
A
87
−14.243
35.195
−7.585
1.00
18.43
C


ATOM
663
NE
ARG
A
87
−14.578
35.661
−6.249
1.00
11.89
N


ATOM
664
CZ
ARG
A
87
−15.736
35.459
−5.638
1.00
12.44
C


ATOM
665
NH1
ARG
A
87
−16.707
34.782
−6.235
1.00
13.57
N


ATOM
666
NH2
ARG
A
87
−15.916
35.950
−4.414
1.00
13.60
N


ATOM
667
N
MET
A
88
−11.172
32.232
−5.059
1.00
9.91
N


ATOM
668
CA
MET
A
88
−10.380
31.028
−4.820
1.00
11.60
C


ATOM
669
C
MET
A
88
−9.109
31.274
−4.009
1.00
12.66
C


ATOM
670
O
MET
A
88
−8.019
30.791
−4.335
1.00
13.08
O


ATOM
671
CB
MET
A
88
−10.068
30.294
−6.129
1.00
13.27
C


ATOM
672
CG
MET
A
88
−11.227
29.455
−6.646
1.00
23.14
C


ATOM
673
SD
MET
A
88
−10.720
28.240
−7.892
1.00
20.98
S


ATOM
674
CE
MET
A
88
−9.588
27.226
−6.961
1.00
30.54
C


ATOM
675
N
VAL
A
89
−9.285
32.044
−2.942
1.00
9.61
N


ATOM
676
CA
AVAL
A
89
−8.252
32.187
−1.938
0.50
8.87
C


ATOM
677
C
VAL
A
89
−8.935
32.155
−0.579
1.00
8.58
C


ATOM
678
O
VAL
A
89
−10.128
32.442
−0.452
1.00
10.43
O


ATOM
679
CB
AVAL
A
89
−7.439
33.484
−2.102
0.50
20.29
C


ATOM
680
CG1
AVAL
A
89
−6.728
33.502
−3.451
0.50
21.26
C


ATOM
681
CG2
AVAL
A
89
−8.335
34.689
−1.951
0.50
11.60
C


ATOM
682
CA
BVAL
A
89
−8.264
32.316
−1.942
0.50
15.07
C


ATOM
683
CB
BVAL
A
89
−7.752
33.771
−2.069
0.50
13.11
C


ATOM
684
CG1
BVAL
A
89
−7.120
34.254
−0.767
0.50
15.31
C


ATOM
685
CG2
BVAL
A
89
−6.786
33.900
−3.241
0.50
28.13
C


ATOM
686
N
VAL
A
90
−8.184
31.737
0.427
1.00
9.08
N


ATOM
687
CA
VAL
A
90
−8.738
31.651
1.774
1.00
11.86
C


ATOM
688
C
VAL
A
90
−7.583
31.817
2.759
1.00
11.67
C


ATOM
689
O
VAL
A
90
−6.506
31.267
2.556
1.00
10.65
O


ATOM
690
CB
VAL
A
90
−9.530
30.322
1.992
1.00
9.75
C


ATOM
691
CG1
VAL
A
90
−8.584
29.132
2.112
1.00
10.71
C


ATOM
692
CG2
VAL
A
90
−10.449
30.428
3.209
1.00
12.69
C


ATOM
693
N
SER
A
91
−7.793
32.610
3.807
1.00
11.86
N


ATOM
694
CA
SER
A
91
−6.708
32.895
4.745
1.00
11.26
C


ATOM
695
C
SER
A
91
−6.179
31.607
5.380
1.00
10.82
C


ATOM
696
O
SER
A
91
−4.972
31.401
5.489
1.00
13.74
O


ATOM
697
CB
SER
A
91
−7.170
33.884
5.817
1.00
14.65
C


ATOM
698
OG
SER
A
91
−8.265
33.367
6.549
1.00
16.21
O


ATOM
699
N
ASP
A
92
−7.105
30.743
5.789
1.00
10.07
N


ATOM
700
CA
ASP
A
92
−6.784
29.407
6.285
1.00
9.45
C


ATOM
701
C
ASP
A
92
−7.983
28.523
5.981
1.00
8.51
C


ATOM
702
O
ASP
A
92
−9.130
28.967
6.055
1.00
8.70
O


ATOM
703
CB
ASP
A
92
−6.516
29.407
7.795
1.00
12.63
C


ATOM
704
CG
ASP
A
92
−6.132
28.016
8.322
1.00
12.62
C


ATOM
705
OD1
ASP
A
92
−4.934
27.663
8.272
1.00
15.77
O


ATOM
706
OD2
ASP
A
92
−7.030
27.276
8.782
1.00
13.34
O


ATOM
707
N
PHE
A
93
−7.714
27.266
5.644
1.00
7.96
N


ATOM
708
CA
PHE
A
93
−8.783
26.351
5.260
1.00
9.18
C


ATOM
709
C
PHE
A
93
−9.900
26.284
6.304
1.00
7.89
C


ATOM
710
O
PHE
A
93
−11.059
26.057
5.975
1.00
8.19
O


ATOM
711
CB
PHE
A
93
−8.233
24.949
4.997
1.00
8.72
C


ATOM
712
CG
PHE
A
93
−9.171
24.080
4.204
1.00
8.08
C


ATOM
713
CD1
PHE
A
93
−9.243
24.196
2.822
1.00
9.70
C


ATOM
714
CD2
PHE
A
93
−9.994
23.163
4.834
1.00
9.05
C


ATOM
715
CE1
PHE
A
93
−10.117
23.401
2.082
1.00
8.73
C


ATOM
716
CE2
PHE
A
93
−10.870
22.363
4.105
1.00
9.40
C


ATOM
717
CZ
PHE
A
93
−10.936
22.488
2.726
1.00
8.70
C


ATOM
718
N
HIS
A
94
−9.546
26.468
7.572
1.00
7.76
N


ATOM
719
CA
HIS
A
94
−10.545
26.363
8.621
1.00
9.12
C


ATOM
720
C
HIS
A
94
−11.690
27.367
8.476
1.00
6.64
C


ATOM
721
O
HIS
A
94
−12.782
27.138
8.982
1.00
8.47
O


ATOM
722
CB
HIS
A
94
−9.922
26.487
10.014
1.00
10.83
C


ATOM
723
CG
HIS
A
94
−10.891
26.175
11.107
1.00
12.48
C


ATOM
724
ND1
HIS
A
94
−11.497
27.153
11.868
1.00
18.24
N


ATOM
725
CD2
HIS
A
94
−11.407
24.996
11.526
1.00
14.58
C


ATOM
726
CE1
HIS
A
94
−12.323
26.586
12.729
1.00
18.04
C


ATOM
727
NE2
HIS
A
94
−12.289
25.278
12.541
1.00
20.93
N


ATOM
728
N
VAL
A
95
−11.433
28.476
7.790
1.00
6.79
N


ATOM
729
CA
VAL
A
95
−12.482
29.445
7.525
1.00
7.84
C


ATOM
730
C
VAL
A
95
−13.713
28.751
6.923
1.00
5.99
C


ATOM
731
O
VAL
A
95
−14.852
29.036
7.299
1.00
7.00
O


ATOM
732
CB
VAL
A
95
−11.981
30.569
6.580
1.00
9.46
C


ATOM
733
CG1
VAL
A
95
−13.146
31.338
5.972
1.00
10.88
C


ATOM
734
CG2
VAL
A
95
−11.040
31.504
7.333
1.00
10.22
C


ATOM
735
N
PHE
A
96
−13.482
27.834
5.976
1.00
5.97
N


ATOM
736
CA
PHE
A
96
−14.606
27.148
5.328
1.00
6.34
C


ATOM
737
C
PHE
A
96
−15.432
26.338
6.323
1.00
4.91
C


ATOM
738
O
PHE
A
96
−16.661
26.310
6.262
1.00
6.56
O


ATOM
739
CB
PHE
A
96
−14.124
26.191
4.231
1.00
7.50
C


ATOM
740
CG
PHE
A
96
−13.463
26.855
3.057
1.00
6.47
C


ATOM
741
CD1
PHE
A
96
−13.997
27.991
2.453
1.00
7.28
C


ATOM
742
CD2
PHE
A
96
−12.332
26.286
2.512
1.00
7.30
C


ATOM
743
CE1
PHE
A
96
−13.378
28.572
1.333
1.00
8.59
C


ATOM
744
CE2
PHE
A
96
−11.707
26.847
1.388
1.00
8.91
C


ATOM
745
CZ
PHE
A
96
−12.237
27.993
0.794
1.00
7.27
C


ATOM
746
N
VAL
A
97
−14.732
25.648
7.228
1.00
6.11
N


ATOM
747
CA
VAL
A
97
−15.375
24.815
8.242
1.00
6.27
C


ATOM
748
C
VAL
A
97
−16.135
25.682
9.259
1.00
5.36
C


ATOM
749
O
VAL
A
97
−17.277
25.403
9.594
1.00
6.07
O


ATOM
750
CB
VAL
A
97
−14.312
23.943
8.952
1.00
7.37
C


ATOM
751
CG1
VAL
A
97
−14.941
23.145
10.091
1.00
10.36
C


ATOM
752
CG2
VAL
A
97
−13.620
23.008
7.943
1.00
9.21
C


ATOM
753
N
ARG
A
98
−15.480
26.734
9.740
1.00
6.54
N


ATOM
754
CA
ARG
A
98
−16.117
27.695
10.638
1.00
7.12
C


ATOM
755
C
ARG
A
98
−17.430
28.221
10.051
1.00
5.89
C


ATOM
756
O
ARG
A
98
−18.456
28.299
10.723
1.00
6.29
O


ATOM
757
CB
ARG
A
98
−15.142
28.850
10.852
1.00
8.63
C


ATOM
758
CG
ARG
A
98
−15.637
29.977
11.756
1.00
10.60
C


ATOM
759
CD
ARG
A
98
−14.744
31.189
11.573
1.00
10.59
C


ATOM
760
NE
ARG
A
98
−15.017
31.792
10.268
1.00
10.15
N


ATOM
761
CZ
ARG
A
98
−14.335
32.796
9.731
1.00
10.10
C


ATOM
762
NH1
ARG
A
98
−13.310
33.337
10.377
1.00
11.94
N


ATOM
763
NH2
ARG
A
98
−14.697
33.265
8.543
1.00
10.42
N


ATOM
764
N
ASP
A
99
−17.395
28.585
8.770
1.00
6.39
N


ATOM
765
CA
ASP
A
99
−18.579
29.151
8.127
1.00
7.52
C


ATOM
766
C
ASP
A
99
−19.709
28.118
7.947
1.00
4.77
C


ATOM
767
O
ASP
A
99
−20.882
28.438
8.121
1.00
5.81
O


ATOM
768
CB
ASP
A
99
−18.211
29.818
6.795
1.00
7.19
C


ATOM
769
CG
ASP
A
99
−17.358
31.083
6.974
1.00
9.39
C


ATOM
770
OD1
ASP
A
99
−17.061
31.490
8.129
1.00
9.24
O


ATOM
771
OD2
ASP
A
99
−16.950
31.677
5.948
1.00
8.90
O


ATOM
772
N
VAL
A
100
−19.349
26.875
7.611
1.00
5.99
N


ATOM
773
CA
VAL
A
100
−20.360
25.815
7.575
1.00
5.66
C


ATOM
774
C
VAL
A
100
−21.017
25.659
8.958
1.00
4.09
C


ATOM
775
O
VAL
A
100
−22.228
25.583
9.061
1.00
6.12
O


ATOM
776
CB
VAL
A
100
−19.783
24.457
7.079
1.00
5.14
C


ATOM
777
CG1
VAL
A
100
−20.807
23.349
7.288
1.00
8.00
C


ATOM
778
CG2
VAL
A
100
−19.404
24.560
5.599
1.00
8.22
C


ATOM
779
N
LEU
A
101
−20.184
25.628
10.001
1.00
5.14
N


ATOM
780
CA
LEU
A
101
−20.722
25.484
11.361
1.00
6.56
C


ATOM
781
C
LEU
A
101
−21.636
26.651
11.768
1.00
6.23
C


ATOM
782
O
LEU
A
101
−22.652
26.444
12.419
1.00
6.91
O


ATOM
783
CB
LEU
A
101
−19.591
25.278
12.369
1.00
7.60
C


ATOM
784
CG
LEU
A
101
−18.909
23.905
12.286
1.00
9.07
C


ATOM
785
CD1
LEU
A
101
−17.675
23.904
13.156
1.00
13.36
C


ATOM
786
CD2
LEU
A
101
−19.878
22.797
12.691
1.00
12.74
C


ATOM
787
N
GLN
A
102
−21.294
27.864
11.342
1.00
5.43
N


ATOM
788
CA
GLN
A
102
−22.177
28.997
11.599
1.00
5.19
C


ATOM
789
C
GLN
A
102
−23.543
28.772
10.970
1.00
6.32
C


ATOM
790
O
GLN
A
102
−24.579
28.956
11.610
1.00
6.82
O


ATOM
791
CB
GLN
A
102
−21.552
30.286
11.068
1.00
6.74
C


ATOM
792
CG
GLN
A
102
−22.446
31.499
11.276
1.00
7.84
C


ATOM
793
CD
GLN
A
102
−21.965
32.740
10.550
1.00
7.29
C


ATOM
794
OE1
GLN
A
102
−20.883
32.767
9.977
1.00
9.47
O


ATOM
795
NE2
GLN
A
102
−22.787
33.791
10.585
1.00
8.18
N


ATOM
796
N
HIS
A
103
−23.552
28.370
9.699
1.00
5.94
N


ATOM
797
CA
HIS
A
103
−24.801
28.171
9.002
1.00
6.08
C


ATOM
798
C
HIS
A
103
−25.598
27.012
9.607
1.00
4.80
C


ATOM
799
O
HIS
A
103
−26.801
27.111
9.797
1.00
6.26
O


ATOM
800
CB
HIS
A
103
−24.531
27.925
7.515
1.00
6.90
C


ATOM
801
CG
HIS
A
103
−25.677
28.278
6.631
1.00
7.96
C


ATOM
802
ND1
HIS
A
103
−25.565
28.282
5.256
1.00
8.59
N


ATOM
803
CD2
HIS
A
103
−26.945
28.655
6.910
1.00
6.64
C


ATOM
804
CE1
HIS
A
103
−26.719
28.655
4.728
1.00
8.29
C


ATOM
805
NE2
HIS
A
103
−27.581
28.872
5.708
1.00
9.02
N


ATOM
806
N
VAL
A
104
−24.913
25.903
9.888
1.00
5.95
N


ATOM
807
CA
VAL
A
104
−25.605
24.765
10.486
1.00
7.58
C


ATOM
808
C
VAL
A
104
−26.224
25.153
11.843
1.00
5.80
C


ATOM
809
O
VAL
A
104
−27.385
24.851
12.108
1.00
7.78
O


ATOM
810
CB
VAL
A
104
−24.653
23.572
10.668
1.00
7.05
C


ATOM
811
CG1
VAL
A
104
−25.302
22.506
11.558
1.00
10.12
C


ATOM
812
CG2
VAL
A
104
−24.250
22.984
9.309
1.00
9.57
C


ATOM
813
N
ASP
A
105
−25.434
25.831
12.676
1.00
6.23
N


ATOM
814
CA
ASP
A
105
−25.933
26.244
13.994
1.00
6.14
C


ATOM
815
C
ASP
A
105
−27.133
27.178
13.856
1.00
8.33
C


ATOM
816
O
ASP
A
105
−28.112
27.052
14.592
1.00
8.56
O


ATOM
817
CB
ASP
A
105
−24.839
26.905
14.843
1.00
7.42
C


ATOM
818
CG
ASP
A
105
−23.776
25.920
15.336
1.00
13.92
C


ATOM
819
OD1
ASP
A
105
−24.061
24.704
15.406
1.00
16.13
O


ATOM
820
OD2
ASP
A
105
−22.648
26.361
15.663
1.00
13.33
O


ATOM
821
N
SER
A
106
−27.082
28.111
12.903
1.00
7.27
N


ATOM
822
CA
ASER
A
106
−28.184
29.047
12.712
0.33
8.06
C


ATOM
823
C
SER
A
106
−29.453
28.338
12.241
1.00
9.29
C


ATOM
824
O
SER
A
106
−30.557
28.661
12.670
1.00
12.36
O


ATOM
825
CB
ASER
A
106
−27.791
30.152
11.727
0.33
12.36
C


ATOM
826
OG
ASER
A
106
−28.878
31.033
11.504
0.33
11.64
O


ATOM
827
CA
BSER
A
106
−28.189
29.044
12.739
0.33
7.72
C


ATOM
828
CB
BSER
A
106
−27.798
30.224
11.836
0.33
8.33
C


ATOM
829
OG
BSER
A
106
−27.502
29.800
10.519
0.33
9.09
O


ATOM
830
CA
CSER
A
106
−28.187
29.046
12.697
0.33
8.20
C


ATOM
831
CB
CSER
A
106
−27.805
30.112
11.671
0.33
14.56
C


ATOM
832
OG
CSER
A
106
−26.705
30.866
12.126
0.33
10.99
O


ATOM
833
N
MET
A
107
−29.301
27.365
11.344
1.00
7.49
N


ATOM
834
CA
AMET
A
107
−30.442
26.604
10.845
0.50
10.61
C


ATOM
835
C
MET
A
107
−31.031
25.708
11.936
1.00
6.77
C


ATOM
836
O
MET
A
107
−32.255
25.615
12.087
1.00
11.21
O


ATOM
837
CB
AMET
A
107
−30.024
25.765
9.629
0.50
11.62
C


ATOM
838
CG
AMET
A
107
−31.142
24.962
8.981
0.50
11.62
C


ATOM
839
SD
AMET
A
107
−32.524
25.957
8.382
0.50
17.80
S


ATOM
840
CE
AMET
A
107
−31.659
27.299
7.589
0.50
16.49
C


ATOM
841
CA
BMET
A
107
−30.454
26.617
10.859
0.50
6.28
C


ATOM
842
CB
BMET
A
107
−30.076
25.798
9.625
0.50
11.75
C


ATOM
843
CG
BMET
A
107
−29.649
26.659
8.460
0.50
12.02
C


ATOM
844
SD
BMET
A
107
−30.974
27.746
7.914
0.50
26.78
S


ATOM
845
CE
BMET
A
107
−32.093
26.557
7.189
0.50
15.11
C


ATOM
846
N
GLN
A
108
−30.154
25.044
12.688
1.00
9.49
N


ATOM
847
CA
AGLN
A
108
−30.611
24.118
13.720
0.50
9.57
C


ATOM
848
C
GLN
A
108
−31.458
24.812
14.782
1.00
11.46
C


ATOM
849
O
GLN
A
108
−32.441
24.243
15.262
1.00
12.43
O


ATOM
850
CB
AGLN
A
108
−29.439
23.367
14.357
0.50
12.58
C


ATOM
851
CG
AGLN
A
108
−28.938
22.209
13.504
0.50
12.65
C


ATOM
852
CD
AGLN
A
108
−28.137
21.196
14.294
0.50
68.54
C


ATOM
853
OE1
AGLN
A
108
−28.661
20.161
14.699
0.50
23.41
O


ATOM
854
NE2
AGLN
A
108
−26.864
21.489
14.521
0.50
7.83
N


ATOM
855
CA
BGLN
A
108
−30.589
24.119
13.737
0.50
8.69
C


ATOM
856
CB
BGLN
A
108
−29.391
23.463
14.430
0.50
9.25
C


ATOM
857
CG
BGLN
A
108
−29.780
22.432
15.483
0.50
13.14
C


ATOM
858
CD
BGLN
A
108
−30.122
21.075
14.882
0.50
70.97
C


ATOM
859
OE1
BGLN
A
108
−29.236
20.263
14.616
0.50
17.19
O


ATOM
860
NE2
BGLN
A
108
−31.413
20.820
14.679
0.50
9.82
N


ATOM
861
N
LYS
A
109
−31.099
26.046
15.133
1.00
10.35
N


ATOM
862
CA
LYS
A
109
−31.860
26.757
16.166
1.00
12.18
C


ATOM
863
C
LYS
A
109
−33.260
27.117
15.670
1.00
15.37
C


ATOM
864
O
LYS
A
109
−34.181
27.259
16.477
1.00
15.02
O


ATOM
865
CB
LYS
A
109
−31.119
28.010
16.647
1.00
12.62
C


ATOM
866
CG
LYS
A
109
−31.207
29.149
15.643
1.00
24.20
C


ATOM
867
CD
LYS
A
109
−30.473
30.401
16.074
1.00
26.44
C


ATOM
868
CE
LYS
A
109
−30.940
31.602
15.256
1.00
17.15
C


ATOM
869
NZ
LYS
A
109
−31.147
31.297
13.808
1.00
22.59
N


ATOM
870
N
ASP
A
110
−33.420
27.265
14.352
1.00
11.64
N


ATOM
871
CA
ASP
A
110
−34.731
27.502
13.748
1.00
14.09
C


ATOM
872
C
ASP
A
110
−35.540
26.202
13.650
1.00
17.43
C


ATOM
873
O
ASP
A
110
−36.768
26.227
13.596
1.00
16.72
O


ATOM
874
CB
ASP
A
110
−34.603
28.125
12.349
1.00
16.62
C


ATOM
875
CG
ASP
A
110
−33.858
29.459
12.349
1.00
35.51
C


ATOM
876
OD1
ASP
A
110
−33.892
30.183
13.363
1.00
27.26
O


ATOM
877
OD2
ASP
A
110
−33.242
29.795
11.314
1.00
33.71
O


ATOM
878
N
TYR
A
111
−34.838
25.070
13.614
1.00
12.88
N


ATOM
879
CA
TYR
A
111
−35.463
23.749
13.495
1.00
11.95
C


ATOM
880
C
TYR
A
111
−34.929
22.816
14.570
1.00
14.36
C


ATOM
881
O
TYR
A
111
−34.283
21.805
14.270
1.00
15.37
O


ATOM
882
CB
TYR
A
111
−35.195
23.153
12.104
1.00
14.49
C


ATOM
883
CG
TYR
A
111
−35.903
23.906
11.010
1.00
11.25
C


ATOM
884
CD1
TYR
A
111
−35.303
24.996
10.389
1.00
11.70
C


ATOM
885
CD2
TYR
A
111
−37.177
23.536
10.605
1.00
13.97
C


ATOM
886
CE1
TYR
A
111
−35.964
25.702
9.397
1.00
12.73
C


ATOM
887
CE2
TYR
A
111
−37.842
24.229
9.609
1.00
13.65
C


ATOM
888
CZ
TYR
A
111
−37.234
25.306
9.013
1.00
13.00
C


ATOM
889
OH
TYR
A
111
−37.903
25.994
8.027
1.00
17.94
O


ATOM
890
N
PRO
A
112
−35.184
23.159
15.842
1.00
16.22
N


ATOM
891
CA
PRO
A
112
−34.592
22.419
16.956
1.00
17.66
C


ATOM
892
C
PRO
A
112
−35.069
20.974
16.951
1.00
18.24
C


ATOM
893
O
PRO
A
112
−36.225
20.702
16.616
1.00
23.82
O


ATOM
894
CB
PRO
A
112
−35.132
23.166
18.190
1.00
24.32
C


ATOM
895
CG
PRO
A
112
−36.365
23.857
17.695
1.00
22.23
C


ATOM
896
CD
PRO
A
112
−35.988
24.298
16.318
1.00
19.06
C


ATOM
897
N
GLY
A
113
−34.176
20.058
17.295
1.00
21.11
N


ATOM
898
CA
GLY
A
113
−34.537
18.657
17.369
1.00
27.87
C


ATOM
899
C
GLY
A
113
−34.396
17.874
16.075
1.00
37.60
C


ATOM
900
O
GLY
A
113
−34.388
16.645
16.105
1.00
26.73
O


ATOM
901
N
LEU
A
114
−34.294
18.556
14.937
1.00
17.32
N


ATOM
902
CA
LEU
A
114
−34.147
17.831
13.678
1.00
16.13
C


ATOM
903
C
LEU
A
114
−32.712
17.349
13.481
1.00
14.38
C


ATOM
904
O
LEU
A
114
−31.757
18.081
13.758
1.00
14.82
O


ATOM
905
CB
LEU
A
114
−34.560
18.689
12.485
1.00
15.76
C


ATOM
906
CG
LEU
A
114
−36.039
19.009
12.274
1.00
19.83
C


ATOM
907
CD1
LEU
A
114
−36.244
19.522
10.860
1.00
22.33
C


ATOM
908
CD2
LEU
A
114
−36.907
17.787
12.537
1.00
26.81
C


ATOM
909
N
PRO
A
115
−32.559
16.110
12.993
1.00
14.51
N


ATOM
910
CA
PRO
A
115
−31.241
15.578
12.631
1.00
14.94
C


ATOM
911
C
PRO
A
115
−30.640
16.390
11.484
1.00
9.64
C


ATOM
912
O
PRO
A
115
−31.386
16.881
10.636
1.00
12.08
O


ATOM
913
CB
PRO
A
115
−31.554
14.159
12.140
1.00
16.54
C


ATOM
914
CG
PRO
A
115
−32.911
13.846
12.664
1.00
25.57
C


ATOM
915
CD
PRO
A
115
−33.640
15.148
12.735
1.00
14.61
C


ATOM
916
N
VAL
A
116
−29.320
16.522
11.472
1.00
8.33
N


ATOM
917
CA
AVAL
A
116
−28.656
17.245
10.396
0.50
6.55
C


ATOM
918
C
VAL
A
116
−27.608
16.371
9.703
1.00
9.54
C


ATOM
919
O
VAL
A
116
−26.774
15.730
10.337
1.00
9.56
O


ATOM
920
CB
AVAL
A
116
−28.055
18.584
10.888
0.50
13.27
C


ATOM
921
CG1
AVAL
A
116
−27.021
18.342
11.977
0.50
10.54
C


ATOM
922
CG2
AVAL
A
116
−27.472
19.385
9.721
0.50
6.54
C


ATOM
923
CA
BVAL
A
116
−28.606
17.265
10.430
0.50
11.46
C


ATOM
924
CB
BVAL
A
116
−27.833
18.463
11.022
0.50
7.27
C


ATOM
925
CG1
BVAL
A
116
−26.972
19.144
9.954
0.50
10.48
C


ATOM
926
CG2
BVAL
A
116
−28.798
19.447
11.664
0.50
11.77
C


ATOM
927
N
PHE
A
117
−27.694
16.350
8.377
1.00
8.25
N


ATOM
928
CA
PHE
A
117
−26.766
15.613
7.523
1.00
6.73
C


ATOM
929
C
PHE
A
117
−25.872
16.596
6.781
1.00
7.65
C


ATOM
930
O
PHE
A
117
−26.221
17.782
6.646
1.00
7.95
O


ATOM
931
CB
PHE
A
117
−27.544
14.791
6.495
1.00
7.19
C


ATOM
932
CG
PHE
A
117
−28.342
13.659
7.089
1.00
9.66
C


ATOM
933
CD1
PHE
A
117
−29.578
13.888
7.675
1.00
9.86
C


ATOM
934
CD2
PHE
A
117
−27.858
12.366
7.035
1.00
11.39
C


ATOM
935
CE1
PHE
A
117
−30.315
12.846
8.221
1.00
11.17
C


ATOM
936
CE2
PHE
A
117
−28.592
11.316
7.576
1.00
12.02
C


ATOM
937
CZ
PHE
A
117
−29.810
11.552
8.167
1.00
12.26
C


ATOM
938
N
LEU
A
118
−24.751
16.093
6.272
1.00
6.39
N


ATOM
939
CA
LEU
A
118
−23.844
16.851
5.409
1.00
6.79
C


ATOM
940
C
LEU
A
118
−23.696
16.104
4.084
1.00
9.58
C


ATOM
941
O
LEU
A
118
−23.659
14.869
4.053
1.00
10.15
O


ATOM
942
CB
LEU
A
118
−22.467
16.990
6.054
1.00
8.39
C


ATOM
943
CG
LEU
A
118
−22.423
17.678
7.421
1.00
7.65
C


ATOM
944
CD1
LEU
A
118
−21.069
17.514
8.065
1.00
12.05
C


ATOM
945
CD2
LEU
A
118
−22.805
19.163
7.274
1.00
14.26
C


ATOM
946
N
LEU
A
119
−23.606
16.848
2.989
1.00
6.75
N


ATOM
947
CA
LEU
A
119
−23.262
16.262
1.695
1.00
6.84
C


ATOM
948
C
LEU
A
119
−22.159
17.111
1.091
1.00
6.93
C


ATOM
949
O
LEU
A
119
−22.288
18.341
1.039
1.00
9.68
O


ATOM
950
CB
LEU
A
119
−24.478
16.235
0.780
1.00
9.51
C


ATOM
951
CG
LEU
A
119
−24.279
15.572
−0.595
1.00
8.88
C


ATOM
952
CD1
LEU
A
119
−25.576
14.929
−1.069
1.00
14.75
C


ATOM
953
CD2
LEU
A
119
−23.738
16.566
−1.628
1.00
12.64
C


ATOM
954
N
GLY
A
120
−21.076
16.486
0.640
1.00
5.97
N


ATOM
955
CA
GLY
A
120
−19.986
17.265
0.088
1.00
6.20
C


ATOM
956
C
GLY
A
120
−19.321
16.567
−1.078
1.00
7.47
C


ATOM
957
O
GLY
A
120
−19.260
15.334
−1.112
1.00
8.48
O


ATOM
958
N
HIS
A
121
−18.805
17.354
−2.013
1.00
4.99
N


ATOM
959
CA
HIS
A
121
−18.051
16.840
−3.158
1.00
5.24
C


ATOM
960
C
HIS
A
121
−16.656
17.442
−3.172
1.00
4.93
C


ATOM
961
O
HIS
A
121
−16.494
18.663
−3.004
1.00
6.29
O


ATOM
962
CB
HIS
A
121
−18.791
17.217
−4.456
1.00
6.08
C


ATOM
963
CG
HIS
A
121
−17.985
17.032
−5.712
1.00
7.22
C


ATOM
964
ND1
HIS
A
121
−17.405
15.831
−6.075
1.00
8.06
N


ATOM
965
CD2
HIS
A
121
−17.688
17.906
−6.707
1.00
6.53
C


ATOM
966
CE1
HIS
A
121
−16.785
15.977
−7.238
1.00
8.61
C


ATOM
967
NE2
HIS
A
121
−16.941
17.227
−7.641
1.00
7.27
N


ATOM
968
N
SER
A
122
−15.635
16.614
−3.358
1.00
5.44
N


ATOM
969
CA
SER
A
122
−14.288
17.105
−3.623
1.00
6.10
C


ATOM
970
C
SER
A
122
−13.779
17.939
−2.431
1.00
7.18
C


ATOM
971
O
SER
A
122
−13.808
17.447
−1.292
1.00
6.60
O


ATOM
972
CB
SER
A
122
−14.240
17.852
−4.976
1.00
7.68
C


ATOM
973
OG
SER
A
122
−12.925
17.935
−5.480
1.00
8.60
O


ATOM
974
N
MET
A
123
−13.298
19.163
−2.648
1.00
7.05
N


ATOM
975
CA
MET
A
123
−12.930
20.012
−1.510
1.00
6.85
C


ATOM
976
C
MET
A
123
−14.067
20.114
−0.498
1.00
5.95
C


ATOM
977
O
MET
A
123
−13.825
20.180
0.715
1.00
6.61
O


ATOM
978
CB
MET
A
123
−12.544
21.418
−1.969
1.00
6.44
C


ATOM
979
CG
MET
A
123
−12.132
22.321
−0.808
1.00
6.74
C


ATOM
980
SD
MET
A
123
−11.872
24.035
−1.346
1.00
9.51
S


ATOM
981
CE
MET
A
123
−13.527
24.679
−1.392
1.00
11.77
C


ATOM
982
N
GLY
A
124
−15.294
20.162
−0.991
1.00
5.76
N


ATOM
983
CA
GLY
A
124
−16.465
20.242
−0.134
1.00
5.33
C


ATOM
984
C
GLY
A
124
−16.644
19.005
0.733
1.00
6.28
C


ATOM
985
O
GLY
A
124
−17.250
19.080
1.807
1.00
7.72
O


ATOM
986
N
GLY
A
125
−16.131
17.871
0.258
1.00
6.88
N


ATOM
987
CA
GLY
A
125
−16.123
16.647
1.048
1.00
6.32
C


ATOM
988
C
GLY
A
125
−15.096
16.702
2.164
1.00
7.06
C


ATOM
989
O
GLY
A
125
−15.362
16.219
3.275
1.00
8.26
O


ATOM
990
N
ALA
A
126
−13.936
17.291
1.906
1.00
6.65
N


ATOM
991
CA
ALA
A
126
−12.967
17.542
2.966
1.00
7.07
C


ATOM
992
C
ALA
A
126
−13.560
18.468
4.027
1.00
8.51
C


ATOM
993
O
ALA
A
126
−13.390
18.234
5.232
1.00
8.99
O


ATOM
994
CB
ALA
A
126
−11.685
18.140
2.394
1.00
9.00
C


ATOM
995
N
ILE
A
127
−14.256
19.510
3.587
1.00
5.80
N


ATOM
996
CA
ILE
A
127
−14.899
20.425
4.512
1.00
5.81
C


ATOM
997
C
ILE
A
127
−15.919
19.663
5.359
1.00
7.22
C


ATOM
998
O
ILE
A
127
−15.978
19.846
6.586
1.00
8.40
O


ATOM
999
CB
ILE
A
127
−15.560
21.585
3.757
1.00
5.20
C


ATOM
1000
CG1
ILE
A
127
−14.479
22.471
3.125
1.00
6.80
C


ATOM
1001
CG2
ILE
A
127
−16.455
22.395
4.692
1.00
7.18
C


ATOM
1002
CD1
ILE
A
127
−14.991
23.467
2.072
1.00
8.65
C


ATOM
1003
N
ALA
A
128
−16.708
18.794
4.735
1.00
6.54
N


ATOM
1004
CA
ALA
A
128
−17.685
17.983
5.472
1.00
6.61
C


ATOM
1005
C
ALA
A
128
−17.014
17.098
6.525
1.00
6.98
C


ATOM
1006
O
ALA
A
128
−17.470
17.038
7.676
1.00
8.44
O


ATOM
1007
CB
ALA
A
128
−18.527
17.138
4.522
1.00
8.15
C


ATOM
1008
N
ILE
A
129
−15.951
16.402
6.138
1.00
6.43
N


ATOM
1009
CA
ILE
A
129
−15.234
15.541
7.073
1.00
8.20
C


ATOM
1010
C
ILE
A
129
−14.742
16.355
8.274
1.00
7.24
C


ATOM
1011
O
ILE
A
129
−14.921
15.943
9.432
1.00
9.40
O


ATOM
1012
CB
ILE
A
129
−14.056
14.835
6.372
1.00
7.21
C


ATOM
1013
CG1
ILE
A
129
−14.577
13.788
5.374
1.00
8.59
C


ATOM
1014
CG2
ILE
A
129
−13.099
14.196
7.392
1.00
8.65
C


ATOM
1015
CD1
ILE
A
129
−13.518
13.302
4.381
1.00
8.43
C


ATOM
1016
N
LEU
A
130
−14.109
17.494
8.016
1.00
7.88
N


ATOM
1017
CA
LEU
A
130
−13.564
18.322
9.090
1.00
7.26
C


ATOM
1018
C
LEU
A
130
−14.648
18.930
9.968
1.00
8.38
C


ATOM
1019
O
LEU
A
130
−14.440
19.099
11.175
1.00
11.78
O


ATOM
1020
CB
LEU
A
130
−12.629
19.396
8.522
1.00
7.43
C


ATOM
1021
CG
LEU
A
130
−11.379
18.824
7.851
1.00
7.20
C


ATOM
1022
CD1
LEU
A
130
−10.588
19.932
7.164
1.00
11.26
C


ATOM
1023
CD2
LEU
A
130
−10.499
18.088
8.856
1.00
11.11
C


ATOM
1024
N
THR
A
131
−15.792
19.247
9.381
1.00
6.76
N


ATOM
1025
CA
THR
A
131
−16.937
19.765
10.120
1.00
7.15
C


ATOM
1026
C
THR
A
131
−17.469
18.709
11.090
1.00
10.49
C


ATOM
1027
O
THR
A
131
−17.694
18.990
12.278
1.00
10.09
O


ATOM
1028
CB
THR
A
131
−18.044
20.221
9.142
1.00
6.73
C


ATOM
1029
OG1
THR
A
131
−17.544
21.303
8.342
1.00
9.73
O


ATOM
1030
CG2
THR
A
131
−19.293
20.695
9.876
1.00
9.04
C


ATOM
1031
N
ALA
A
132
−17.667
17.493
10.591
1.00
8.54
N


ATOM
1032
CA
ALA
A
132
−18.145
16.401
11.429
1.00
10.74
C


ATOM
1033
C
ALA
A
132
−17.152
16.098
12.545
1.00
13.98
C


ATOM
1034
O
ALA
A
132
−17.559
15.799
13.674
1.00
17.35
O


ATOM
1035
CB
ALA
A
132
−18.395
15.153
10.588
1.00
12.21
C


ATOM
1036
N
ALA
A
133
−15.862
16.190
12.247
1.00
11.80
N


ATOM
1037
CA
ALA
A
133
−14.836
15.861
13.228
1.00
13.45
C


ATOM
1038
C
ALA
A
133
−14.746
16.916
14.325
1.00
17.15
C


ATOM
1039
O
ALA
A
133
−14.327
16.619
15.451
1.00
19.66
O


ATOM
1040
CB
ALA
A
133
−13.475
15.669
12.546
1.00
15.60
C


ATOM
1041
N
GLU
A
134
−15.139
18.143
14.001
1.00
16.89
N


ATOM
1042
CA
GLU
A
134
−15.107
19.240
14.961
1.00
17.80
C


ATOM
1043
C
GLU
A
134
−16.272
19.188
15.952
1.00
21.56
C


ATOM
1044
O
GLU
A
134
−16.207
19.805
17.023
1.00
21.34
O


ATOM
1045
CB
GLU
A
134
−15.086
20.585
14.232
1.00
19.70
C


ATOM
1046
CG
GLU
A
134
−15.042
21.779
15.166
1.00
31.66
C


ATOM
1047
CD
GLU
A
134
−14.422
22.997
14.524
1.00
77.78
C


ATOM
1048
OE1
GLU
A
134
−13.675
22.834
13.534
1.00
29.11
O


ATOM
1049
OE2
GLU
A
134
−14.678
24.116
15.017
1.00
32.70
O


ATOM
1050
N
ARG
A
135
−17.328
18.457
15.597
1.00
15.43
N


ATOM
1051
CA
ARG
A
135
−18.489
18.264
16.470
1.00
14.20
C


ATOM
1052
C
ARG
A
135
−18.795
16.777
16.633
1.00
15.41
C


ATOM
1053
O
ARG
A
135
−19.804
16.274
16.129
1.00
16.46
O


ATOM
1054
CB
ARG
A
135
−19.717
18.994
15.924
1.00
15.89
C


ATOM
1055
CG
ARG
A
135
−19.520
20.485
15.716
1.00
19.46
C


ATOM
1056
CD
ARG
A
135
−19.647
21.282
17.015
1.00
22.44
C


ATOM
1057
NE
ARG
A
135
−19.498
22.713
16.762
1.00
17.05
N


ATOM
1058
CZ
ARG
A
135
−20.503
23.518
16.428
1.00
13.79
C


ATOM
1059
NH1
ARG
A
135
−21.738
23.043
16.315
1.00
15.86
N


ATOM
1060
NH2
ARG
A
135
−20.264
24.802
16.195
1.00
15.95
N


ATOM
1061
N
PRO
A
136
−17.921
16.062
17.353
1.00
20.51
N


ATOM
1062
CA
PRO
A
136
−18.056
14.607
17.494
1.00
24.54
C


ATOM
1063
C
PRO
A
136
−19.441
14.177
17.975
1.00
29.40
C


ATOM
1064
O
PRO
A
136
−19.924
14.677
18.994
1.00
23.65
O


ATOM
1065
CB
PRO
A
136
−17.007
14.262
18.554
1.00
27.33
C


ATOM
1066
CG
PRO
A
136
−16.000
15.361
18.471
1.00
33.77
C


ATOM
1067
CD
PRO
A
136
−16.774
16.596
18.109
1.00
22.45
C


ATOM
1068
N
GLY
A
137
−20.063
13.257
17.244
1.00
19.62
N


ATOM
1069
CA
GLY
A
137
−21.346
12.689
17.613
1.00
16.56
C


ATOM
1070
C
GLY
A
137
−22.563
13.519
17.260
1.00
18.57
C


ATOM
1071
O
GLY
A
137
−23.687
13.134
17.559
1.00
24.49
O


ATOM
1072
N
HIS
A
138
−22.347
14.657
16.605
1.00
14.73
N


ATOM
1073
CA
HIS
A
138
−23.423
15.610
16.384
1.00
14.81
C


ATOM
1074
C
HIS
A
138
−24.243
15.369
15.103
1.00
13.75
C


ATOM
1075
O
HIS
A
138
−25.472
15.419
15.122
1.00
17.01
O


ATOM
1076
CB
HIS
A
138
−22.853
17.027
16.385
1.00
17.64
C


ATOM
1077
CG
HIS
A
138
−23.888
18.087
16.192
1.00
19.02
C


ATOM
1078
ND1
HIS
A
138
−24.835
18.385
17.150
1.00
24.37
N


ATOM
1079
CD2
HIS
A
138
−24.131
18.918
15.151
1.00
21.19
C


ATOM
1080
CE1
HIS
A
138
−25.615
19.354
16.706
1.00
53.43
C


ATOM
1081
NE2
HIS
A
138
−25.210
19.695
15.496
1.00
28.81
N


ATOM
1082
N
PHE
A
139
−23.557
15.105
13.993
1.00
12.98
N


ATOM
1083
CA
PHE
A
139
−24.235
14.934
12.708
1.00
12.74
C


ATOM
1084
C
PHE
A
139
−24.824
13.537
12.536
1.00
8.88
C


ATOM
1085
O
PHE
A
139
−24.244
12.555
13.007
1.00
12.86
O


ATOM
1086
CB
PHE
A
139
−23.280
15.293
11.558
1.00
11.40
C


ATOM
1087
CG
PHE
A
139
−22.933
16.751
11.522
1.00
8.97
C


ATOM
1088
CD1
PHE
A
139
−23.765
17.647
10.875
1.00
10.65
C


ATOM
1089
CD2
PHE
A
139
−21.807
17.228
12.169
1.00
10.08
C


ATOM
1090
CE1
PHE
A
139
−23.456
19.002
10.856
1.00
11.55
C


ATOM
1091
CE2
PHE
A
139
−21.501
18.580
12.162
1.00
9.46
C


ATOM
1092
CZ
PHE
A
139
−22.330
19.464
11.501
1.00
11.90
C


ATOM
1093
N
ALA
A
140
−25.969
13.462
11.872
1.00
10.10
N


ATOM
1094
CA
ALA
A
140
−26.681
12.201
11.680
1.00
11.80
C


ATOM
1095
C
ALA
A
140
−26.034
11.356
10.589
1.00
12.36
C


ATOM
1096
O
ALA
A
140
−26.193
10.135
10.557
1.00
13.34
O


ATOM
1097
CB
ALA
A
140
−28.128
12.459
11.333
1.00
12.22
C


ATOM
1098
N
GLY
A
141
−25.327
12.009
9.675
1.00
9.31
N


ATOM
1099
CA
GLY
A
141
−24.774
11.289
8.546
1.00
9.56
C


ATOM
1100
C
GLY
A
141
−24.130
12.212
7.548
1.00
11.98
C


ATOM
1101
O
GLY
A
141
−24.391
13.420
7.525
1.00
10.18
O


ATOM
1102
N
MET
A
142
−23.292
11.614
6.711
1.00
8.32
N


ATOM
1103
CA
AMET
A
142
−22.502
12.331
5.725
0.70
8.58
C


ATOM
1104
C
MET
A
142
−22.554
11.573
4.389
1.00
9.98
C


ATOM
1105
O
MET
A
142
−22.320
10.356
4.367
1.00
10.51
O


ATOM
1106
CB
AMET
A
142
−21.067
12.360
6.237
0.70
25.98
C


ATOM
1107
CG
AMET
A
142
−20.126
13.326
5.579
0.70
16.80
C


ATOM
1108
SD
AMET
A
142
−18.699
13.543
6.679
0.70
15.44
S


ATOM
1109
CE
AMET
A
142
−18.032
11.891
6.773
0.70
13.72
C


ATOM
1110
CA
BMET
A
142
−22.589
12.353
5.680
0.30
8.38
C


ATOM
1111
CB
BMET
A
142
−21.163
12.652
6.104
0.30
4.61
C


ATOM
1112
CG
BMET
A
142
−21.010
13.930
6.885
0.30
26.62
C


ATOM
1113
SD
BMET
A
142
−19.322
14.124
7.459
0.30
21.03
S


ATOM
1114
CE
BMET
A
142
−18.407
13.324
6.145
0.30
22.67
C


ATOM
1115
N
VAL
A
143
−22.825
12.281
3.299
1.00
7.01
N


ATOM
1116
CA
VAL
A
143
−22.730
11.708
1.964
1.00
7.21
C


ATOM
1117
C
VAL
A
143
−21.561
12.389
1.268
1.00
7.22
C


ATOM
1118
O
VAL
A
143
−21.534
13.624
1.135
1.00
7.86
O


ATOM
1119
CB
VAL
A
143
−24.017
11.957
1.159
1.00
8.34
C


ATOM
1120
CG1
VAL
A
143
−23.839
11.460
−0.286
1.00
10.34
C


ATOM
1121
CG2
VAL
A
143
−25.207
11.275
1.833
1.00
10.57
C


ATOM
1122
N
LEU
A
144
−20.576
11.601
0.853
1.00
5.85
N


ATOM
1123
CA
LEU
A
144
−19.357
12.139
0.276
1.00
6.51
C


ATOM
1124
C
LEU
A
144
−19.222
11.661
−1.160
1.00
8.86
C


ATOM
1125
O
LEU
A
144
−19.195
10.456
−1.439
1.00
7.94
O


ATOM
1126
CB
LEU
A
144
−18.143
11.680
1.084
1.00
6.27
C


ATOM
1127
CG
LEU
A
144
−18.117
12.083
2.559
1.00
9.97
C


ATOM
1128
CD1
LEU
A
144
−16.959
11.359
3.232
1.00
11.52
C


ATOM
1129
CD2
LEU
A
144
−17.983
13.615
2.693
1.00
11.05
C


ATOM
1130
N
ILE
A
145
−19.179
12.618
−2.079
1.00
5.85
N


ATOM
1131
CA
ILE
A
145
−18.973
12.315
−3.490
1.00
6.29
C


ATOM
1132
C
ILE
A
145
−17.545
12.689
−3.844
1.00
6.81
C


ATOM
1133
O
ILE
A
145
−17.194
13.876
−3.904
1.00
6.91
O


ATOM
1134
CB
ILE
A
145
−19.957
13.103
−4.377
1.00
5.41
C


ATOM
1135
CG1
ILE
A
145
−21.400
12.836
−3.954
1.00
9.22
C


ATOM
1136
CG2
ILE
A
145
−19.742
12.726
−5.846
1.00
8.27
C


ATOM
1137
CD1
ILE
A
145
−22.431
13.717
−4.651
1.00
9.95
C


ATOM
1138
N
SER
A
146
−16.689
11.689
−4.034
1.00
6.84
N


ATOM
1139
CA
SER
A
146
−15.301
11.934
−4.411
1.00
6.91
C


ATOM
1140
C
SER
A
146
−14.655
12.966
−3.493
1.00
7.51
C


ATOM
1141
O
SER
A
146
−14.114
13.975
−3.958
1.00
7.48
O


ATOM
1142
CB
SER
A
146
−15.201
12.439
−5.857
1.00
8.55
C


ATOM
1143
OG
SER
A
146
−16.035
11.684
−6.705
1.00
19.57
O


ATOM
1144
N
PRO
A
147
−14.668
12.714
−2.178
1.00
7.19
N


ATOM
1145
CA
PRO
A
147
−14.114
13.708
−1.250
1.00
6.02
C


ATOM
1146
C
PRO
A
147
−12.612
13.828
−1.423
1.00
8.82
C


ATOM
1147
O
PRO
A
147
−11.933
12.854
−1.765
1.00
8.83
O


ATOM
1148
CB
PRO
A
147
−14.431
13.108
0.123
1.00
7.28
C


ATOM
1149
CG
PRO
A
147
−14.419
11.598
−0.149
1.00
7.97
C


ATOM
1150
CD
PRO
A
147
−15.119
11.491
−1.486
1.00
7.21
C


ATOM
1151
N
LEU
A
148
−12.077
15.017
−1.183
1.00
7.64
N


ATOM
1152
CA
LEU
A
148
−10.643
15.212
−1.171
1.00
7.57
C


ATOM
1153
C
LEU
A
148
−10.037
14.612
0.100
1.00
10.69
C


ATOM
1154
O
LEU
A
148
−10.082
15.230
1.180
1.00
13.01
O


ATOM
1155
CB
LEU
A
148
−10.308
16.702
−1.295
1.00
10.01
C


ATOM
1156
CG
LEU
A
148
−8.831
17.084
−1.238
1.00
13.54
C


ATOM
1157
CD1
LEU
A
148
−8.050
16.349
−2.318
1.00
18.01
C


ATOM
1158
CD2
LEU
A
148
−8.660
18.592
−1.378
1.00
19.64
C


ATOM
1159
N
VAL
A
149
−9.491
13.403
−0.029
1.00
10.28
N


ATOM
1160
CA
VAL
A
149
−8.837
12.724
1.087
1.00
9.11
C


ATOM
1161
C
VAL
A
149
−7.359
12.529
0.775
1.00
11.36
C


ATOM
1162
O
VAL
A
149
−6.496
12.909
1.560
1.00
13.79
O


ATOM
1163
CB
VAL
A
149
−9.514
11.374
1.410
1.00
7.75
C


ATOM
1164
CG1
VAL
A
149
−8.733
10.636
2.512
1.00
9.82
C


ATOM
1165
CG2
VAL
A
149
−10.957
11.592
1.816
1.00
8.89
C


ATOM
1166
N
LEU
A
150
−7.069
11.934
−0.380
1.00
10.74
N


ATOM
1167
CA
LEU
A
150
−5.705
11.867
−0.886
1.00
12.38
C


ATOM
1168
C
LEU
A
150
−5.711
12.416
−2.295
1.00
13.20
C


ATOM
1169
O
LEU
A
150
−6.533
12.017
−3.119
1.00
14.22
O


ATOM
1170
CB
LEU
A
150
−5.185
10.428
−0.916
1.00
13.51
C


ATOM
1171
CG
LEU
A
150
−4.880
9.789
0.439
1.00
17.14
C


ATOM
1172
CD1
LEU
A
150
−3.807
10.577
1.181
1.00
19.78
C


ATOM
1173
CD2
LEU
A
150
−4.453
8.333
0.248
1.00
21.23
C


ATOM
1174
N
ALA
A
151
−4.803
13.338
−2.572
1.00
12.30
N


ATOM
1175
CA
ALA
A
151
−4.701
13.879
−3.917
1.00
16.69
C


ATOM
1176
C
ALA
A
151
−3.972
12.890
−4.819
1.00
15.92
C


ATOM
1177
O
ALA
A
151
−3.240
12.021
−4.344
1.00
15.20
O


ATOM
1178
CB
ALA
A
151
−3.981
15.220
−3.893
1.00
20.19
C


ATOM
1179
N
ASN
A
152
−4.191
13.011
−6.120
1.00
14.44
N


ATOM
1180
CA
ASN
A
152
−3.363
12.292
−7.071
1.00
13.52
C


ATOM
1181
C
ASN
A
152
−1.896
12.543
−6.728
1.00
14.72
C


ATOM
1182
O
ASN
A
152
−1.469
13.694
−6.634
1.00
15.93
O


ATOM
1183
CB
ASN
A
152
−3.674
12.764
−8.487
1.00
19.76
C


ATOM
1184
CG
ASN
A
152
−2.809
12.089
−9.524
1.00
18.69
C


ATOM
1185
OD1
ASN
A
152
−1.632
12.417
−9.674
1.00
20.74
O


ATOM
1186
ND2
ASN
A
152
−3.389
11.138
−10.251
1.00
23.13
N


ATOM
1187
N
PRO
A
153
−1.122
11.467
−6.513
1.00
16.79
N


ATOM
1188
CA
PRO
A
153
0.264
11.579
−6.041
1.00
17.04
C


ATOM
1189
C
PRO
A
153
1.151
12.434
−6.946
1.00
16.59
C


ATOM
1190
O
PRO
A
153
1.901
13.272
−6.445
1.00
17.20
O


ATOM
1191
CB
PRO
A
153
0.749
10.125
−6.034
1.00
23.14
C


ATOM
1192
CG
PRO
A
153
−0.491
9.322
−5.850
1.00
26.11
C


ATOM
1193
CD
PRO
A
153
−1.570
10.065
−6.589
1.00
18.51
C


ATOM
1194
N
GLU
A
154
1.072
12.222
−8.254
1.00
17.01
N


ATOM
1195
CA
GLU
A
154
1.860
13.019
−9.191
1.00
17.94
C


ATOM
1196
C
GLU
A
154
1.463
14.494
−9.151
1.00
16.34
C


ATOM
1197
O
GLU
A
154
2.329
15.369
−9.141
1.00
16.77
O


ATOM
1198
CB
GLU
A
154
1.754
12.464
−10.616
1.00
19.72
C


ATOM
1199
CG
GLU
A
154
2.514
11.160
−10.810
1.00
56.06
C


ATOM
1200
CD
GLU
A
154
2.713
10.806
−12.270
1.00
91.32
C


ATOM
1201
OE1
GLU
A
154
2.125
11.488
−13.136
1.00
55.91
O


ATOM
1202
OE2
GLU
A
154
3.462
9.845
−12.549
1.00
63.05
O


ATOM
1203
N
SER
A
155
0.161
14.767
−9.124
1.00
14.06
N


ATOM
1204
CA
SER
A
155
−0.333
16.141
−9.044
1.00
13.04
C


ATOM
1205
C
SER
A
155
0.097
16.835
−7.752
1.00
13.20
C


ATOM
1206
O
SER
A
155
0.483
18.007
−7.764
1.00
15.06
O


ATOM
1207
CB
SER
A
155
−1.856
16.180
−9.150
1.00
19.70
C


ATOM
1208
OG
SER
A
155
−2.293
15.617
−10.374
1.00
24.81
O


ATOM
1209
N
ALA
A
156
0.018
16.113
−6.641
1.00
13.13
N


ATOM
1210
CA
ALA
A
156
0.429
16.669
−5.357
1.00
14.25
C


ATOM
1211
C
ALA
A
156
1.913
17.014
−5.368
1.00
14.95
C


ATOM
1212
O
ALA
A
156
2.321
18.091
−4.916
1.00
13.46
O


ATOM
1213
CB
ALA
A
156
0.118
15.696
−4.224
1.00
16.70
C


ATOM
1214
N
THR
A
157
2.723
16.098
−5.890
1.00
13.19
N


ATOM
1215
CA
THR
A
157
4.164
16.321
−5.953
1.00
12.51
C


ATOM
1216
C
THR
A
157
4.494
17.494
−6.868
1.00
11.38
C


ATOM
1217
O
THR
A
157
5.311
18.351
−6.521
1.00
13.00
O


ATOM
1218
CB
THR
A
157
4.897
15.066
−6.435
1.00
16.00
C


ATOM
1219
OG1
THR
A
157
4.623
13.991
−5.527
1.00
20.31
O


ATOM
1220
CG2
THR
A
157
6.400
15.307
−6.485
1.00
16.90
C


ATOM
1221
N
THR
A
158
3.858
17.525
−8.036
1.00
11.66
N


ATOM
1222
CA
THR
A
158
4.036
18.639
−8.965
1.00
12.58
C


ATOM
1223
C
THR
A
158
3.720
19.944
−8.250
1.00
12.22
C


ATOM
1224
O
THR
A
158
4.459
20.927
−8.360
1.00
11.34
O


ATOM
1225
CB
THR
A
158
3.119
18.502
−10.194
1.00
13.07
C


ATOM
1226
OG1
THR
A
158
3.539
17.373
−10.972
1.00
16.60
O


ATOM
1227
CG2
THR
A
158
3.174
19.755
−11.065
1.00
12.95
C


ATOM
1228
N
PHE
A
159
2.620
19.962
−7.506
1.00
10.44
N


ATOM
1229
CA
PHE
A
159
2.245
21.202
−6.850
1.00
11.72
C


ATOM
1230
C
PHE
A
159
3.243
21.604
−5.765
1.00
9.72
C


ATOM
1231
O
PHE
A
159
3.589
22.777
−5.652
1.00
11.95
O


ATOM
1232
CB
PHE
A
159
0.842
21.161
−6.256
1.00
10.51
C


ATOM
1233
CG
PHE
A
159
0.509
22.417
−5.524
1.00
12.93
C


ATOM
1234
CD1
PHE
A
159
0.146
23.552
−6.224
1.00
12.61
C


ATOM
1235
CD2
PHE
A
159
0.658
22.492
−4.150
1.00
13.42
C


ATOM
1236
CE1
PHE
A
159
−0.113
24.742
−5.566
1.00
15.13
C


ATOM
1237
CE2
PHE
A
159
0.399
23.682
−3.479
1.00
12.40
C


ATOM
1238
CZ
PHE
A
159
0.013
24.805
−4.189
1.00
11.69
C


ATOM
1239
N
LYS
A
160
3.699
20.645
−4.969
1.00
11.04
N


ATOM
1240
CA
LYS
A
160
4.644
20.969
−3.906
1.00
13.94
C


ATOM
1241
C
LYS
A
160
5.937
21.531
−4.483
1.00
14.48
C


ATOM
1242
O
LYS
A
160
6.520
22.463
−3.936
1.00
14.62
O


ATOM
1243
CB
LYS
A
160
4.933
19.743
−3.041
1.00
14.06
C


ATOM
1244
CG
LYS
A
160
3.750
19.302
−2.200
1.00
16.18
C


ATOM
1245
CD
LYS
A
160
4.049
18.014
−1.454
1.00
19.05
C


ATOM
1246
CE
LYS
A
160
2.930
17.694
−0.473
1.00
30.54
C


ATOM
1247
NZ
LYS
A
160
3.310
16.609
0.472
1.00
58.66
N


ATOM
1248
N
VAL
A
161
6.391
20.961
−5.591
1.00
13.06
N


ATOM
1249
CA
VAL
A
161
7.601
21.457
−6.237
1.00
17.55
C


ATOM
1250
C
VAL
A
161
7.391
22.878
−6.766
1.00
15.74
C


ATOM
1251
O
VAL
A
161
8.248
23.749
−6.605
1.00
14.78
O


ATOM
1252
CB
VAL
A
161
8.044
20.524
−7.372
1.00
14.83
C


ATOM
1253
CG1
VAL
A
161
9.160
21.173
−8.186
1.00
19.24
C


ATOM
1254
CG2
VAL
A
161
8.494
19.183
−6.797
1.00
18.97
C


ATOM
1255
N
LEU
A
162
6.239
23.110
−7.380
1.00
11.62
N


ATOM
1256
CA
LEU
A
162
5.875
24.434
−7.867
1.00
10.78
C


ATOM
1257
C
LEU
A
162
5.815
25.443
−6.726
1.00
15.67
C


ATOM
1258
O
LEU
A
162
6.360
26.543
−6.826
1.00
14.46
O


ATOM
1259
CB
LEU
A
162
4.520
24.381
−8.572
1.00
14.81
C


ATOM
1260
CG
LEU
A
162
3.973
25.727
−9.046
1.00
22.23
C


ATOM
1261
CD1
LEU
A
162
4.800
26.249
−10.206
1.00
23.30
C


ATOM
1262
CD2
LEU
A
162
2.507
25.604
−9.439
1.00
24.52
C


ATOM
1263
N
ALA
A
163
5.148
25.070
−5.639
1.00
11.29
N


ATOM
1264
CA
ALA
A
163
4.999
25.983
−4.509
1.00
14.76
C


ATOM
1265
C
ALA
A
163
6.362
26.338
−3.922
1.00
14.91
C


ATOM
1266
O
ALA
A
163
6.602
27.491
−3.560
1.00
16.78
O


ATOM
1267
CB
ALA
A
163
4.086
25.382
−3.442
1.00
14.03
C


ATOM
1268
N
ALA
A
164
7.250
25.351
−3.835
1.00
13.62
N


ATOM
1269
CA
ALA
A
164
8.598
25.582
−3.322
1.00
18.16
C


ATOM
1270
C
ALA
A
164
9.400
26.525
−4.226
1.00
23.79
C


ATOM
1271
O
ALA
A
164
10.109
27.402
−3.732
1.00
18.76
O


ATOM
1272
CB
ALA
A
164
9.331
24.262
−3.137
1.00
20.61
C


ATOM
1273
N
LYS
A
165
9.297
26.339
−5.544
1.00
17.69
N


ATOM
1274
CA
LYS
A
165
9.950
27.243
−6.494
1.00
23.27
C


ATOM
1275
C
LYS
A
165
9.456
28.657
−6.279
1.00
23.11
C


ATOM
1276
O
LYS
A
165
10.238
29.600
−6.157
1.00
33.57
O


ATOM
1277
CB
LYS
A
165
9.604
26.885
−7.940
1.00
30.74
C


ATOM
1278
CG
LYS
A
165
10.148
25.590
−8.486
1.00
58.56
C


ATOM
1279
CD
LYS
A
165
10.057
25.660
−10.004
1.00
28.67
C


ATOM
1280
CE
LYS
A
165
9.648
24.348
−10.632
1.00
63.54
C


ATOM
1281
NZ
LYS
A
165
9.245
24.586
−12.048
1.00
25.56
N


ATOM
1282
N
VAL
A
166
8.137
28.800
−6.273
1.00
19.68
N


ATOM
1283
CA
VAL
A
166
7.504
30.102
−6.156
1.00
19.36
C


ATOM
1284
C
VAL
A
166
7.910
30.804
−4.862
1.00
38.58
C


ATOM
1285
O
VAL
A
166
8.095
32.019
−4.840
1.00
41.10
O


ATOM
1286
CB
VAL
A
166
5.966
29.992
−6.274
1.00
23.38
C


ATOM
1287
CG1
VAL
A
166
5.300
31.297
−5.869
1.00
33.39
C


ATOM
1288
CG2
VAL
A
166
5.576
29.597
−7.698
1.00
19.99
C


ATOM
1289
N
LEU
A
167
8.069
30.031
−3.793
1.00
30.39
N


ATOM
1290
CA
LEU
A
167
8.493
30.578
−2.504
1.00
29.35
C


ATOM
1291
C
LEU
A
167
10.005
30.798
−2.400
1.00
37.42
C


ATOM
1292
O
LEU
A
167
10.451
31.869
−1.989
1.00
45.82
O


ATOM
1293
CB
LEU
A
167
8.033
29.670
−1.359
1.00
36.36
C


ATOM
1294
CG
LEU
A
167
8.823
29.820
−0.053
1.00
72.40
C


ATOM
1295
CD1
LEU
A
167
8.542
31.166
0.601
1.00
72.35
C


ATOM
1296
CD2
LEU
A
167
8.525
28.677
0.910
1.00
57.03
C


ATOM
1297
N
ASN
A
168
10.787
29.784
−2.765
1.00
27.55
N


ATOM
1298
CA
ASN
A
168
12.237
29.810
−2.569
1.00
27.38
C


ATOM
1299
C
ASN
A
168
13.020
30.567
−3.639
1.00
50.32
C


ATOM
1300
O
ASN
A
168
14.246
30.674
−3.565
1.00
38.60
O


ATOM
1301
CB
ASN
A
168
12.791
28.387
−2.456
1.00
31.18
C


ATOM
1302
CG
ASN
A
168
12.355
27.689
−1.185
1.00
60.71
C


ATOM
1303
OD1
ASN
A
168
12.552
26.483
−1.028
1.00
56.08
O


ATOM
1304
ND2
ASN
A
168
11.759
28.443
−0.268
1.00
64.87
N


ATOM
1305
N
SER
A
169
12.313
31.077
−4.639
1.00
23.88
N


ATOM
1306
CA
SER
A
169
12.939
31.844
−5.708
1.00
29.73
C


ATOM
1307
C
SER
A
169
12.086
33.057
−6.050
1.00
22.64
C


ATOM
1308
CB
SER
A
169
13.123
30.976
−6.957
1.00
35.25
C


ATOM
1309
OG
SER
A
169
13.964
29.866
−6.694
1.00
55.90
O


ATOM
1310
O
SER
A
169
12.359
33.753
−7.029
1.00
27.12
O


ATOM
1311
N
VAL
A
170
11.059
33.298
−5.237
1.00
20.95
N


ATOM
1312
CA
VAL
A
170
10.077
34.351
−5.489
1.00
17.05
C


ATOM
1313
C
VAL
A
170
9.765
34.510
−6.985
1.00
23.05
C


ATOM
1314
O
VAL
A
170
9.865
35.603
−7.545
1.00
22.27
O


ATOM
1315
CB
VAL
A
170
10.503
35.708
−4.858
1.00
23.73
C


ATOM
1316
CG1
VAL
A
170
10.500
35.607
−3.335
1.00
22.54
C


ATOM
1317
CG2
VAL
A
170
11.874
36.134
−5.362
1.00
28.79
C


ATOM
1318
N
LEU
A
171
9.395
33.406
−7.632
1.00
19.61
N


ATOM
1319
CA
LEU
A
171
8.996
33.451
−9.036
1.00
17.49
C


ATOM
1320
C
LEU
A
171
7.615
34.085
−9.147
1.00
17.41
C


ATOM
1321
O
LEU
A
171
6.686
33.667
−8.453
1.00
19.36
O


ATOM
1322
CB
LEU
A
171
8.963
32.047
−9.636
1.00
18.63
C


ATOM
1323
CG
LEU
A
171
10.282
31.276
−9.665
1.00
26.27
C


ATOM
1324
CD1
LEU
A
171
10.099
29.942
−10.378
1.00
25.23
C


ATOM
1325
CD2
LEU
A
171
11.364
32.100
−10.339
1.00
22.36
C


ATOM
1326
N
PRO
A
172
7.471
35.094
−10.022
1.00
14.88
N


ATOM
1327
CA
PRO
A
172
6.223
35.853
−10.159
1.00
14.86
C


ATOM
1328
C
PRO
A
172
5.302
35.268
−11.227
1.00
19.81
C


ATOM
1329
O
PRO
A
172
5.760
34.501
−12.068
1.00
19.14
O


ATOM
1330
CB
PRO
A
172
6.715
37.221
−10.627
1.00
20.22
C


ATOM
1331
CG
PRO
A
172
7.938
36.910
−11.435
1.00
18.21
C


ATOM
1332
CD
PRO
A
172
8.543
35.636
−10.880
1.00
14.89
C


ATOM
1333
N
ASN
A
173
4.024
35.633
−11.184
1.00
22.85
N


ATOM
1334
CA
ASN
A
173
3.073
35.266
−12.235
1.00
29.29
C


ATOM
1335
C
ASN
A
173
3.027
33.769
−12.528
1.00
31.64
C


ATOM
1336
O
ASN
A
173
2.795
33.356
−13.665
1.00
39.41
O


ATOM
1337
CB
ASN
A
173
3.386
36.033
−13.520
1.00
32.47
C


ATOM
1338
CG
ASN
A
173
3.475
37.531
−13.296
1.00
85.40
C


ATOM
1339
OD1
ASN
A
173
4.280
38.218
−13.926
1.00
59.68
O


ATOM
1340
ND2
ASN
A
173
2.652
38.044
−12.388
1.00
49.78
N


ATOM
1341
N
LEU
A
174
3.262
32.962
−11.499
1.00
20.60
N


ATOM
1342
CA
LEU
A
174
3.201
31.512
−11.623
1.00
18.27
C


ATOM
1343
C
LEU
A
174
2.155
30.946
−10.685
1.00
25.26
C


ATOM
1344
O
LEU
A
174
2.226
31.128
−9.472
1.00
24.16
O


ATOM
1345
CB
LEU
A
174
4.550
30.872
−11.296
1.00
24.24
C


ATOM
1346
CG
LEU
A
174
5.479
30.472
−12.438
1.00
55.63
C


ATOM
1347
CD1
LEU
A
174
6.625
29.655
−11.870
1.00
27.72
C


ATOM
1348
CD2
LEU
A
174
4.722
29.681
−13.495
1.00
33.38
C


ATOM
1349
N
SER
A
175
1.178
30.253
−11.247
1.00
19.56
N


ATOM
1350
CA
SER
A
175
0.212
29.552
−10.427
1.00
21.54
C


ATOM
1351
C
SER
A
175
0.218
28.100
−10.855
1.00
20.84
C


ATOM
1352
O
SER
A
175
0.958
27.711
−11.761
1.00
18.86
O


ATOM
1353
CB
SER
A
175
−1.182
30.157
−10.597
1.00
21.73
C


ATOM
1354
OG
SER
A
175
−1.679
29.929
−11.903
1.00
27.11
O


ATOM
1355
N
SER
A
176
−0.599
27.294
−10.192
1.00
16.51
N


ATOM
1356
CA
SER
A
176
−0.839
25.940
−10.640
1.00
17.04
C


ATOM
1357
C
SER
A
176
−1.830
26.028
−11.798
1.00
16.54
C


ATOM
1358
CB
SER
A
176
−1.400
25.104
−9.485
1.00
17.23
C


ATOM
1359
OG
SER
A
176
−1.399
23.728
−9.797
1.00
20.69
O


ATOM
1360
O
SER
A
176
−2.331
27.110
−12.112
1.00
18.57
O


ATOM
1361
N
GLY
A
177
−2.104
24.904
−12.451
1.00
17.84
N


ATOM
1362
CA
GLY
A
177
−3.036
24.906
−13.564
1.00
18.83
C


ATOM
1363
C
GLY
A
177
−4.467
25.206
−13.144
1.00
14.12
C


ATOM
1364
O
GLY
A
177
−4.815
25.120
−11.960
1.00
14.14
O


ATOM
1365
N
PRO
A
178
−5.312
25.585
−14.105
1.00
13.09
N


ATOM
1366
CA
PRO
A
178
−6.729
25.736
−13.773
1.00
11.60
C


ATOM
1367
C
PRO
A
178
−7.390
24.370
−13.676
1.00
12.12
C


ATOM
1368
CB
PRO
A
178
−7.290
26.513
−14.962
1.00
13.40
C


ATOM
1369
CG
PRO
A
178
−6.386
26.154
−16.109
1.00
22.97
C


ATOM
1370
CD
PRO
A
178
−5.029
25.841
−15.532
1.00
16.39
C


ATOM
1371
O
PRO
A
178
−6.763
23.345
−13.972
1.00
14.28
O


ATOM
1372
N
ILE
A
179
−8.640
24.359
−13.244
1.00
11.60
N


ATOM
1373
CA
ILE
A
179
−9.404
23.126
−13.199
1.00
11.12
C


ATOM
1374
C
ILE
A
179
−9.693
22.662
−14.633
1.00
13.21
C


ATOM
1375
O
ILE
A
179
−10.218
23.434
−15.444
1.00
14.90
O


ATOM
1376
CB
ILE
A
179
−10.707
23.357
−12.430
1.00
10.34
C


ATOM
1377
CG1
ILE
A
179
−10.392
23.610
−10.955
1.00
14.08
C


ATOM
1378
CG2
ILE
A
179
−11.661
22.179
−12.590
1.00
14.20
C


ATOM
1379
CD1
ILE
A
179
−11.518
24.273
−10.208
1.00
12.85
C


ATOM
1380
N
ASP
A
180
−9.324
21.421
−14.957
1.00
12.91
N


ATOM
1381
CA
ASP
A
180
−9.567
20.876
−16.302
1.00
10.51
C


ATOM
1382
C
ASP
A
180
−10.792
19.985
−16.253
1.00
10.15
C


ATOM
1383
O
ASP
A
180
−10.707
18.794
−15.928
1.00
12.02
O


ATOM
1384
CB
ASP
A
180
−8.352
20.098
−16.819
1.00
15.37
C


ATOM
1385
CG
ASP
A
180
−8.568
19.517
−18.225
1.00
13.01
C


ATOM
1386
OD1
ASP
A
180
−9.687
19.646
−18.779
1.00
13.99
O


ATOM
1387
OD2
ASP
A
180
−7.603
18.924
−18.759
1.00
17.65
O


ATOM
1388
N
SER
A
181
−11.938
20.570
−16.553
1.00
10.15
N


ATOM
1389
CA
SER
A
181
−13.199
19.873
−16.436
1.00
8.88
C


ATOM
1390
C
SER
A
181
−13.310
18.686
−17.396
1.00
8.77
C


ATOM
1391
O
SER
A
181
−14.090
17.763
−17.151
1.00
9.07
O


ATOM
1392
CB
SER
A
181
−14.345
20.842
−16.676
1.00
9.62
C


ATOM
1393
OG
SER
A
181
−14.145
21.506
−17.921
1.00
11.79
O


ATOM
1394
N
SER
A
182
−12.554
18.727
−18.494
1.00
8.53
N


ATOM
1395
CA
SER
A
182
−12.675
17.689
−19.518
1.00
8.81
C


ATOM
1396
C
SER
A
182
−12.171
16.328
−19.032
1.00
9.43
C


ATOM
1397
O
SER
A
182
−12.534
15.295
−19.607
1.00
11.89
O


ATOM
1398
CB
SER
A
182
−11.943
18.093
−20.807
1.00
10.60
C


ATOM
1399
OG
SER
A
182
−10.552
17.939
−20.670
1.00
14.76
O


ATOM
1400
N
VAL
A
183
−11.337
16.319
−17.996
1.00
8.40
N


ATOM
1401
CA
VAL
A
183
−10.853
15.054
−17.451
1.00
9.30
C


ATOM
1402
C
VAL
A
183
−11.542
14.715
−16.136
1.00
7.84
C


ATOM
1403
O
VAL
A
183
−11.115
13.791
−15.443
1.00
9.46
O


ATOM
1404
CB
VAL
A
183
−9.327
15.039
−17.257
1.00
9.77
C


ATOM
1405
CG1
VAL
A
183
−8.625
15.275
−18.591
1.00
13.98
C


ATOM
1406
CG2
VAL
A
183
−8.889
16.064
−16.216
1.00
12.72
C


ATOM
1407
N
LEU
A
184
−12.597
15.457
−15.809
1.00
6.49
N


ATOM
1408
CA
LEU
A
184
−13.291
15.282
−14.531
1.00
7.18
C


ATOM
1409
C
LEU
A
184
−14.721
14.767
−14.699
1.00
8.94
C


ATOM
1410
O
LEU
A
184
−15.369
14.390
−13.728
1.00
7.03
O


ATOM
1411
CB
LEU
A
184
−13.225
16.574
−13.686
1.00
7.27
C


ATOM
1412
CG
LEU
A
184
−11.797
17.033
−13.382
1.00
5.81
C


ATOM
1413
CD1
LEU
A
184
−11.784
18.379
−12.656
1.00
8.64
C


ATOM
1414
CD2
LEU
A
184
−11.033
15.993
−12.570
1.00
8.78
C


ATOM
1415
N
SER
A
185
−15.217
14.731
−15.934
1.00
6.60
N


ATOM
1416
CA
SER
A
185
−16.447
14.011
−16.253
1.00
8.04
C


ATOM
1417
C
SER
A
185
−16.374
13.507
−17.684
1.00
9.06
C


ATOM
1418
O
SER
A
185
−15.879
14.221
−18.562
1.00
9.74
O


ATOM
1419
CB
SER
A
185
−17.690
14.892
−16.123
1.00
7.37
C


ATOM
1420
OG
SER
A
185
−18.831
14.192
−16.585
1.00
7.90
O


ATOM
1421
N
ARG
A
186
−16.913
12.318
−17.932
1.00
7.99
N


ATOM
1422
CA
ARG
A
186
−16.986
11.784
−19.296
1.00
8.17
C


ATOM
1423
C
ARG
A
186
−18.178
12.352
−20.056
1.00
11.97
C


ATOM
1424
O
ARG
A
186
−18.309
12.136
−21.264
1.00
11.61
O


ATOM
1425
CB
ARG
A
186
−17.057
10.250
−19.272
1.00
10.81
C


ATOM
1426
CG
ARG
A
186
−15.777
9.591
−18.788
1.00
10.26
C


ATOM
1427
CD
ARG
A
186
−15.960
8.081
−18.583
1.00
10.74
C


ATOM
1428
NE
ARG
A
186
−17.037
7.816
−17.635
1.00
10.08
N


ATOM
1429
CZ
ARG
A
186
−17.587
6.622
−17.426
1.00
13.38
C


ATOM
1430
NH1
ARG
A
186
−17.149
5.566
−18.108
1.00
14.05
N


ATOM
1431
NH2
ARG
A
186
−18.572
6.479
−16.551
1.00
12.05
N


ATOM
1432
N
ASN
A
187
−19.062
13.055
−19.358
1.00
8.13
N


ATOM
1433
CA
ASN
A
187
−20.254
13.610
−19.978
1.00
8.18
C


ATOM
1434
C
ASN
A
187
−19.929
14.967
−20.601
1.00
10.33
C


ATOM
1435
O
ASN
A
187
−19.820
15.977
−19.899
1.00
9.28
O


ATOM
1436
CB
ASN
A
187
−21.383
13.720
−18.943
1.00
7.71
C


ATOM
1437
CG
ASN
A
187
−22.700
14.159
−19.548
1.00
8.65
C


ATOM
1438
OD1
ASN
A
187
−22.731
14.989
−20.462
1.00
10.78
O


ATOM
1439
ND2
ASN
A
187
−23.797
13.613
−19.046
1.00
11.01
N


ATOM
1440
N
LYS
A
188
−19.750
14.982
−21.918
1.00
10.09
N


ATOM
1441
CA
LYS
A
188
−19.290
16.182
−22.611
1.00
10.45
C


ATOM
1442
C
LYS
A
188
−20.274
17.335
−22.497
1.00
8.22
C


ATOM
1443
O
LYS
A
188
−19.854
18.498
−22.440
1.00
10.20
O


ATOM
1444
CB
LYS
A
188
−18.984
15.865
−24.079
1.00
12.84
C


ATOM
1445
CG
LYS
A
188
−17.682
15.087
−24.242
1.00
16.21
C


ATOM
1446
CD
LYS
A
188
−17.316
14.886
−25.702
1.00
30.25
C


ATOM
1447
CE
LYS
A
188
−17.788
13.538
−26.215
1.00
59.11
C


ATOM
1448
NZ
LYS
A
188
−16.968
12.425
−25.656
1.00
62.44
N


ATOM
1449
N
THR
A
189
−21.558
17.018
−22.461
1.00
8.16
N


ATOM
1450
CA
THR
A
189
−22.590
18.029
−22.287
1.00
9.52
C


ATOM
1451
C
THR
A
189
−22.440
18.716
−20.921
1.00
11.14
C


ATOM
1452
O
THR
A
189
−22.499
19.947
−20.822
1.00
10.76
O


ATOM
1453
CB
THR
A
189
−24.002
17.422
−22.419
1.00
11.47
C


ATOM
1454
OG1
THR
A
189
−24.083
16.672
−23.641
1.00
21.86
O


ATOM
1455
CG2
THR
A
189
−25.069
18.514
−22.442
1.00
15.48
C


ATOM
1456
N
GLU
A
190
−22.243
17.917
−19.866
1.00
7.70
N


ATOM
1457
CA
GLU
A
190
−22.042
18.473
−18.525
1.00
6.48
C


ATOM
1458
C
GLU
A
190
−20.762
19.295
−18.456
1.00
6.09
C


ATOM
1459
O
GLU
A
190
−20.723
20.327
−17.779
1.00
7.57
O


ATOM
1460
CB
GLU
A
190
−22.047
17.360
−17.468
1.00
7.78
C


ATOM
1461
CG
GLU
A
190
−23.387
16.671
−17.321
1.00
10.63
C


ATOM
1462
CD
GLU
A
190
−24.469
17.574
−16.735
1.00
19.84
C


ATOM
1463
OE1
GLU
A
190
−24.149
18.687
−16.262
1.00
14.70
O


ATOM
1464
OE2
GLU
A
190
−25.650
17.164
−16.743
1.00
48.07
O


ATOM
1465
N
VAL
A
191
−19.709
18.864
−19.139
1.00
6.34
N


ATOM
1466
CA
VAL
A
191
−18.481
19.653
−19.206
1.00
7.23
C


ATOM
1467
C
VAL
A
191
−18.752
21.037
−19.820
1.00
9.26
C


ATOM
1468
O
VAL
A
191
−18.303
22.054
−19.289
1.00
8.62
O


ATOM
1469
CB
VAL
A
191
−17.369
18.914
−19.990
1.00
7.21
C


ATOM
1470
CG1
VAL
A
191
−16.176
19.816
−20.252
1.00
10.02
C


ATOM
1471
CG2
VAL
A
191
−16.945
17.649
−19.229
1.00
9.40
C


ATOM
1472
N
ASP
A
192
−19.496
21.067
−20.924
1.00
8.63
N


ATOM
1473
CA
ASP
A
192
−19.794
22.326
−21.601
1.00
9.12
C


ATOM
1474
C
ASP
A
192
−20.662
23.232
−20.723
1.00
7.92
C


ATOM
1475
O
ASP
A
192
−20.481
24.456
−20.705
1.00
8.58
O


ATOM
1476
CB
ASP
A
192
−20.505
22.061
−22.930
1.00
8.58
C


ATOM
1477
CG
ASP
A
192
−19.550
21.589
−24.026
1.00
12.97
C


ATOM
1478
OD1
ASP
A
192
−18.343
21.894
−23.950
1.00
19.09
O


ATOM
1479
OD2
ASP
A
192
−20.020
20.912
−24.968
1.00
13.65
O


ATOM
1480
N
ILE
A
193
−21.614
22.636
−20.014
1.00
6.50
N


ATOM
1481
CA
ILE
A
193
−22.479
23.391
−19.111
1.00
6.64
C


ATOM
1482
C
ILE
A
193
−21.658
24.019
−17.992
1.00
8.30
C


ATOM
1483
O
ILE
A
193
−21.797
25.211
−17.684
1.00
9.16
O


ATOM
1484
CB
ILE
A
193
−23.594
22.499
−18.555
1.00
6.90
C


ATOM
1485
CG1
ILE
A
193
−24.608
22.219
−19.667
1.00
9.28
C


ATOM
1486
CG2
ILE
A
193
−24.291
23.153
−17.355
1.00
9.32
C


ATOM
1487
CD1
ILE
A
193
−25.635
21.164
−19.323
1.00
11.81
C


ATOM
1488
N
TYR
A
194
−20.766
23.230
−17.403
1.00
6.21
N


ATOM
1489
CA
TYR
A
194
−19.902
23.749
−16.359
1.00
6.09
C


ATOM
1490
C
TYR
A
194
−19.037
24.896
−16.880
1.00
6.70
C


ATOM
1491
O
TYR
A
194
−18.930
25.955
−16.238
1.00
7.31
O


ATOM
1492
CB
TYR
A
194
−19.023
22.623
−15.814
1.00
6.50
C


ATOM
1493
CG
TYR
A
194
−17.954
23.098
−14.861
1.00
6.88
C


ATOM
1494
CD1
TYR
A
194
−18.239
23.297
−13.501
1.00
7.46
C


ATOM
1495
CD2
TYR
A
194
−16.670
23.355
−15.305
1.00
6.99
C


ATOM
1496
CE1
TYR
A
194
−17.254
23.734
−12.635
1.00
6.75
C


ATOM
1497
CE2
TYR
A
194
−15.681
23.774
−14.445
1.00
8.92
C


ATOM
1498
CZ
TYR
A
194
−15.981
23.965
−13.105
1.00
7.57
C


ATOM
1499
OH
TYR
A
194
−14.961
24.384
−12.276
1.00
11.18
O


ATOM
1500
N
ASN
A
195
−18.401
24.688
−18.028
1.00
6.89
N


ATOM
1501
CA
ASN
A
195
−17.462
25.666
−18.552
1.00
8.53
C


ATOM
1502
C
ASN
A
195
−18.103
27.032
−18.812
1.00
8.54
C


ATOM
1503
O
ASN
A
195
−17.411
28.044
−18.744
1.00
10.97
O


ATOM
1504
CB
ASN
A
195
−16.820
25.175
−19.856
1.00
8.26
C


ATOM
1505
CG
ASN
A
195
−15.794
24.074
−19.648
1.00
13.25
C


ATOM
1506
OD1
ASN
A
195
−15.364
23.433
−20.610
1.00
14.63
O


ATOM
1507
ND2
ASN
A
195
−15.385
23.861
−18.416
1.00
9.36
N


ATOM
1508
N
SER
A
196
−19.388
27.069
−19.120
1.00
6.92
N


ATOM
1509
CA
ASER
A
196
−20.047
28.337
−19.432
0.35
8.60
C


ATOM
1510
C
SER
A
196
−20.869
28.885
−18.264
1.00
8.11
C


ATOM
1511
O
SER
A
196
−21.634
29.832
−18.424
1.00
8.53
O


ATOM
1512
CB
ASER
A
196
−20.917
28.198
−20.679
0.35
15.69
C


ATOM
1513
OG
ASER
A
196
−21.879
27.180
−20.503
0.35
10.54
O


ATOM
1514
CA
BSER
A
196
−20.047
28.337
−19.432
0.35
8.84
C


ATOM
1515
CB
BSER
A
196
−20.917
28.198
−20.679
0.35
15.69
C


ATOM
1516
OG
BSER
A
196
−21.878
27.179
−20.504
0.35
10.54
O


ATOM
1517
CA
CSER
A
196
−20.030
28.350
−19.424
0.30
7.88
C


ATOM
1518
CB
CSER
A
196
−20.829
28.311
−20.741
0.30
17.42
C


ATOM
1519
OG
CSER
A
196
−21.208
26.999
−21.123
0.30
5.94
O


ATOM
1520
N
ASP
A
197
−20.712
28.283
−17.088
1.00
7.36
N


ATOM
1521
CA
ASP
A
197
−21.378
28.788
−15.890
1.00
7.39
C


ATOM
1522
C
ASP
A
197
−20.675
30.091
−15.452
1.00
5.11
C


ATOM
1523
O
ASP
A
197
−19.472
30.105
−15.231
1.00
7.19
O


ATOM
1524
CB
ASP
A
197
−21.307
27.710
−14.799
1.00
7.00
C


ATOM
1525
CG
ASP
A
197
−22.001
28.111
−13.506
1.00
12.88
C


ATOM
1526
OD1
ASP
A
197
−22.176
29.315
−13.216
1.00
9.53
O


ATOM
1527
OD2
ASP
A
197
−22.354
27.193
−12.734
1.00
9.68
O


ATOM
1528
N
PRO
A
198
−21.441
31.197
−15.362
1.00
7.84
N


ATOM
1529
CA
PRO
A
198
−20.779
32.480
−15.077
1.00
8.09
C


ATOM
1530
C
PRO
A
198
−20.238
32.580
−13.644
1.00
9.46
C


ATOM
1531
O
PRO
A
198
−19.519
33.532
−13.335
1.00
12.16
O


ATOM
1532
CB
PRO
A
198
−21.895
33.517
−15.282
1.00
8.81
C


ATOM
1533
CG
PRO
A
198
−23.166
32.783
−15.271
1.00
12.52
C


ATOM
1534
CD
PRO
A
198
−22.891
31.321
−15.544
1.00
8.16
C


ATOM
1535
N
LEU
A
199
−20.561
31.604
−12.796
1.00
7.46
N


ATOM
1536
CA
LEU
A
199
−20.220
31.683
−11.372
1.00
10.37
C


ATOM
1537
C
LEU
A
199
−19.048
30.798
−10.951
1.00
12.05
C


ATOM
1538
O
LEU
A
199
−18.687
30.775
−9.763
1.00
11.02
O


ATOM
1539
CB
LEU
A
199
−21.451
31.367
−10.526
1.00
7.80
C


ATOM
1540
CG
LEU
A
199
−22.611
32.350
−10.675
1.00
10.76
C


ATOM
1541
CD1
LEU
A
199
−23.818
31.905
−9.861
1.00
11.32
C


ATOM
1542
CD2
LEU
A
199
−22.164
33.760
−10.266
1.00
14.15
C


ATOM
1543
N
ILE
A
200
−18.459
30.065
−11.895
1.00
8.07
N


ATOM
1544
CA
ILE
A
200
−17.275
29.255
−11.599
1.00
6.42
C


ATOM
1545
C
ILE
A
200
−16.006
30.084
−11.705
1.00
7.71
C


ATOM
1546
O
ILE
A
200
−16.051
31.229
−12.184
1.00
12.31
O


ATOM
1547
CB
ILE
A
200
−17.172
28.017
−12.531
1.00
7.22
C


ATOM
1548
CG1
ILE
A
200
−16.906
28.441
−13.992
1.00
10.03
C


ATOM
1549
CG2
ILE
A
200
−18.421
27.151
−12.401
1.00
8.89
C


ATOM
1550
CD1
ILE
A
200
−16.361
27.315
−14.869
1.00
10.39
C


ATOM
1551
N
CYS
A
201
−14.875
29.524
−11.293
1.00
7.95
N


ATOM
1552
CA
CYS
A
201
−13.587
30.182
−11.468
1.00
9.49
C


ATOM
1553
C
CYS
A
201
−12.723
29.352
−12.404
1.00
13.48
C


ATOM
1554
O
CYS
A
201
−12.434
28.187
−12.106
1.00
13.84
O


ATOM
1555
CB
CYS
A
201
−12.870
30.343
−10.125
1.00
12.53
C


ATOM
1556
SG
CYS
A
201
−11.270
31.173
−10.289
1.00
17.94
S


ATOM
1557
N
ARG
A
202
−12.303
29.959
−13.515
1.00
11.34
N


ATOM
1558
CA
ARG
A
202
−11.533
29.264
−14.554
1.00
11.13
C


ATOM
1559
C
ARG
A
202
−10.064
29.653
−14.550
1.00
12.74
C


ATOM
1560
O
ARG
A
202
−9.317
29.269
−15.449
1.00
14.37
O


ATOM
1561
CB
ARG
A
202
−12.123
29.558
−15.936
1.00
12.12
C


ATOM
1562
CG
ARG
A
202
−13.617
29.317
−16.013
1.00
15.07
C


ATOM
1563
CD
ARG
A
202
−14.121
29.443
−17.439
1.00
19.32
C


ATOM
1564
NE
ARG
A
202
−13.749
28.272
−18.222
1.00
18.70
N


ATOM
1565
CZ
ARG
A
202
−13.869
28.184
−19.543
1.00
26.00
C


ATOM
1566
NH1
ARG
A
202
−14.349
29.207
−20.236
1.00
22.12
N


ATOM
1567
NH2
ARG
A
202
−13.509
27.071
−20.166
1.00
19.47
N


ATOM
1568
N
ALA
A
203
−9.646
30.408
−13.538
1.00
12.06
N


ATOM
1569
CA
ALA
A
203
−8.263
30.859
−13.438
1.00
13.49
C


ATOM
1570
C
ALA
A
203
−7.346
29.766
−12.897
1.00
12.80
C


ATOM
1571
O
ALA
A
203
−7.812
28.796
−12.294
1.00
12.79
O


ATOM
1572
CB
ALA
A
203
−8.180
32.109
−12.557
1.00
16.80
C


ATOM
1573
N
GLY
A
204
−6.043
29.919
−13.104
1.00
12.71
N


ATOM
1574
CA
GLY
A
204
−5.083
29.005
−12.518
1.00
13.49
C


ATOM
1575
C
GLY
A
204
−5.213
29.001
−11.006
1.00
14.76
C


ATOM
1576
O
GLY
A
204
−5.478
30.037
−10.395
1.00
14.87
O


ATOM
1577
N
LEU
A
205
−5.053
27.838
−10.394
1.00
12.15
N


ATOM
1578
CA
ALEU
A
205
−5.099
27.763
−8.938
0.50
12.52
C


ATOM
1579
C
LEU
A
205
−3.877
28.430
−8.323
1.00
12.12
C


ATOM
1580
O
LEU
A
205
−2.738
28.073
−8.617
1.00
14.37
O


ATOM
1581
CB
ALEU
A
205
−5.233
26.317
−8.455
0.50
21.10
C


ATOM
1582
CG
ALEU
A
205
−6.675
25.832
−8.280
0.50
22.60
C


ATOM
1583
CD1
ALEU
A
205
−7.347
25.614
−9.629
0.50
46.72
C


ATOM
1584
CD2
ALEU
A
205
−6.729
24.565
−7.437
0.50
21.86
C


ATOM
1585
CA
BLEU
A
205
−5.093
27.740
−8.941
0.50
14.43
C


ATOM
1586
CB
BLEU
A
205
−5.152
26.271
−8.524
0.50
15.12
C


ATOM
1587
CG
BLEU
A
205
−5.569
25.980
−7.083
0.50
25.82
C


ATOM
1588
CD1
BLEU
A
205
−6.333
24.666
−6.994
0.50
50.50
C


ATOM
1589
CD2
BLEU
A
205
−4.358
25.965
−6.180
0.50
39.57
C


ATOM
1590
N
LYS
A
206
−4.119
29.419
−7.467
1.00
10.85
N


ATOM
1591
CA
LYS
A
206
−3.027
30.161
−6.853
1.00
11.66
C


ATOM
1592
C
LYS
A
206
−2.265
29.295
−5.866
1.00
12.14
C


ATOM
1593
O
LYS
A
206
−2.852
28.446
−5.188
1.00
11.48
O


ATOM
1594
CB
LYS
A
206
−3.563
31.409
−6.157
1.00
15.47
C


ATOM
1595
CG
LYS
A
206
−4.079
32.451
−7.138
1.00
20.12
C


ATOM
1596
CD
LYS
A
206
−4.787
33.588
−6.426
1.00
37.03
C


ATOM
1597
CE
LYS
A
206
−5.080
34.727
−7.388
1.00
51.15
C


ATOM
1598
NZ
LYS
A
206
−3.843
35.225
−8.052
1.00
92.27
N


ATOM
1599
N
VAL
A
207
−0.959
29.503
−5.774
1.00
10.91
N


ATOM
1600
CA
VAL
A
207
−0.145
28.709
−4.867
1.00
11.61
C


ATOM
1601
C
VAL
A
207
−0.645
28.809
−3.421
1.00
12.70
C


ATOM
1602
O
VAL
A
207
−0.680
27.816
−2.702
1.00
12.20
O


ATOM
1603
CB
VAL
A
207
1.347
29.092
−4.957
1.00
13.57
C


ATOM
1604
CG1
VAL
A
207
2.124
28.508
−3.785
1.00
16.27
C


ATOM
1605
CG2
VAL
A
207
1.918
28.614
−6.280
1.00
15.13
C


ATOM
1606
N
CYS
A
208
−1.043
30.001
−2.988
1.00
10.56
N


ATOM
1607
CA
CYS
A
208
−1.537
30.138
−1.619
1.00
12.51
C


ATOM
1608
C
CYS
A
208
−2.766
29.252
−1.368
1.00
12.06
C


ATOM
1609
O
CYS
A
208
−2.927
28.698
−0.284
1.00
12.63
O


ATOM
1610
CB
CYS
A
208
−1.844
31.602
−1.287
1.00
13.86
C


ATOM
1611
SG
CYS
A
208
−3.147
32.334
−2.291
1.00
19.08
S


ATOM
1612
N
PHE
A
209
−3.620
29.107
−2.378
1.00
10.09
N


ATOM
1613
CA
PHE
A
209
−4.827
28.297
−2.226
1.00
10.18
C


ATOM
1614
C
PHE
A
209
−4.474
26.817
−2.261
1.00
9.52
C


ATOM
1615
O
PHE
A
209
−4.999
26.045
−1.466
1.00
9.70
O


ATOM
1616
CB
PHE
A
209
−5.862
28.632
−3.298
1.00
8.96
C


ATOM
1617
CG
PHE
A
209
−7.213
28.020
−3.047
1.00
10.85
C


ATOM
1618
CD1
PHE
A
209
−7.979
28.421
−1.965
1.00
15.24
C


ATOM
1619
CD2
PHE
A
209
−7.721
27.055
−3.904
1.00
11.35
C


ATOM
1620
CE1
PHE
A
209
−9.230
27.863
−1.737
1.00
19.47
C


ATOM
1621
CE2
PHE
A
209
−8.967
26.493
−3.680
1.00
13.97
C


ATOM
1622
CZ
PHE
A
209
−9.722
26.901
−2.603
1.00
16.68
C


ATOM
1623
N
GLY
A
210
−3.596
26.423
−3.179
1.00
10.17
N


ATOM
1624
CA
GLY
A
210
−3.119
25.054
−3.215
1.00
10.27
C


ATOM
1625
C
GLY
A
210
−2.529
24.619
−1.887
1.00
9.64
C


ATOM
1626
O
GLY
A
210
−2.727
23.483
−1.453
1.00
10.94
O


ATOM
1627
N
ILE
A
211
−1.798
25.525
−1.246
1.00
9.76
N


ATOM
1628
CA
ILE
A
211
−1.219
25.237
0.060
1.00
10.73
C


ATOM
1629
C
ILE
A
211
−2.334
24.946
1.064
1.00
9.63
C


ATOM
1630
O
ILE
A
211
−2.225
24.023
1.872
1.00
11.27
O


ATOM
1631
CB
ILE
A
211
−0.326
26.394
0.547
1.00
10.51
C


ATOM
1632
CG1
ILE
A
211
0.968
26.424
−0.264
1.00
14.10
C


ATOM
1633
CG2
ILE
A
211
−0.019
26.260
2.051
1.00
15.16
C


ATOM
1634
CD1
ILE
A
211
1.777
27.688
−0.060
1.00
19.74
C


ATOM
1635
N
GLN
A
212
−3.415
25.716
0.999
1.00
8.87
N


ATOM
1636
CA
GLN
A
212
−4.550
25.457
1.887
1.00
9.46
C


ATOM
1637
C
GLN
A
212
−5.228
24.114
1.579
1.00
10.55
C


ATOM
1638
O
GLN
A
212
−5.725
23.448
2.492
1.00
10.09
O


ATOM
1639
CB
GLN
A
212
−5.549
26.618
1.865
1.00
8.41
C


ATOM
1640
CG
GLN
A
212
−4.979
27.899
2.496
1.00
9.42
C


ATOM
1641
CD
GLN
A
212
−4.434
27.662
3.902
1.00
10.45
C


ATOM
1642
OE1
GLN
A
212
−5.011
26.906
4.686
1.00
11.66
O


ATOM
1643
NE2
GLN
A
212
−3.323
28.311
4.222
1.00
14.57
N


ATOM
1644
N
LEU
A
213
−5.241
23.704
0.306
1.00
9.40
N


ATOM
1645
CA
LEU
A
213
−5.764
22.378
−0.036
1.00
10.49
C


ATOM
1646
C
LEU
A
213
−4.854
21.266
0.505
1.00
9.28
C


ATOM
1647
O
LEU
A
213
−5.343
20.236
0.962
1.00
10.04
O


ATOM
1648
CB
LEU
A
213
−5.973
22.236
−1.546
1.00
9.58
C


ATOM
1649
CG
LEU
A
213
−7.050
23.140
−2.143
1.00
10.05
C


ATOM
1650
CD1
LEU
A
213
−7.097
22.976
−3.663
1.00
14.31
C


ATOM
1651
CD2
LEU
A
213
−8.407
22.838
−1.528
1.00
14.79
C


ATOM
1652
N
LEU
A
214
−3.541
21.477
0.478
1.00
9.43
N


ATOM
1653
CA
LEU
A
214
−2.616
20.540
1.123
1.00
11.38
C


ATOM
1654
C
LEU
A
214
−2.855
20.488
2.634
1.00
11.40
C


ATOM
1655
O
LEU
A
214
−2.800
19.411
3.249
1.00
11.93
O


ATOM
1656
CB
LEU
A
214
−1.157
20.914
0.856
1.00
12.30
C


ATOM
1657
CG
LEU
A
214
−0.639
20.817
−0.582
1.00
17.43
C


ATOM
1658
CD1
LEU
A
214
0.811
21.288
−0.643
1.00
17.60
C


ATOM
1659
CD2
LEU
A
214
−0.764
19.397
−1.091
1.00
18.65
C


ATOM
1660
N
ASN
A
215
−3.105
21.650
3.234
1.00
10.49
N


ATOM
1661
CA
ASN
A
215
−3.460
21.701
4.651
1.00
11.90
C


ATOM
1662
C
ASN
A
215
−4.717
20.879
4.909
1.00
10.86
C


ATOM
1663
O
ASN
A
215
−4.788
20.130
5.884
1.00
12.49
O


ATOM
1664
CB
ASN
A
215
−3.696
23.143
5.109
1.00
9.81
C


ATOM
1665
CG
ASN
A
215
−2.413
23.943
5.231
1.00
13.81
C


ATOM
1666
OD1
ASN
A
215
−1.309
23.393
5.195
1.00
15.52
O


ATOM
1667
ND2
ASN
A
215
−2.556
25.257
5.381
1.00
15.57
N


ATOM
1668
N
ALA
A
216
−5.709
21.009
4.031
1.00
9.70
N


ATOM
1669
CA
ALA
A
216
−6.940
20.245
4.172
1.00
10.39
C


ATOM
1670
C
ALA
A
216
−6.662
18.742
4.172
1.00
11.63
C


ATOM
1671
O
ALA
A
216
−7.177
18.013
5.021
1.00
11.59
O


ATOM
1672
CB
ALA
A
216
−7.942
20.617
3.078
1.00
11.90
C


ATOM
1673
N
VAL
A
217
−5.856
18.282
3.217
1.00
11.05
N


ATOM
1674
CA
VAL
A
217
−5.522
16.864
3.131
1.00
12.23
C


ATOM
1675
C
VAL
A
217
−4.844
16.383
4.417
1.00
11.43
C


ATOM
1676
O
VAL
A
217
−5.205
15.340
4.961
1.00
11.37
O


ATOM
1677
CB
VAL
A
217
−4.642
16.575
1.898
1.00
12.66
C


ATOM
1678
CG1
VAL
A
217
−4.053
15.157
1.955
1.00
15.90
C


ATOM
1679
CG2
VAL
A
217
−5.449
16.780
0.618
1.00
13.74
C


ATOM
1680
N
SER
A
218
−3.884
17.155
4.910
1.00
10.49
N


ATOM
1681
CA
ASER
A
218
−3.194
16.822
6.152
0.50
11.81
C


ATOM
1682
C
SER
A
218
−4.174
16.752
7.322
1.00
12.18
C


ATOM
1683
O
SER
A
218
−4.142
15.814
8.127
1.00
13.97
O


ATOM
1684
CB
ASER
A
218
−2.103
17.853
6.442
0.50
15.09
C


ATOM
1685
OG
ASER
A
218
−1.031
17.731
5.527
0.50
17.82
O


ATOM
1686
CA
BSER
A
218
−3.194
16.811
6.151
0.50
10.96
C


ATOM
1687
CB
BSER
A
218
−2.072
17.812
6.439
0.50
14.67
C


ATOM
1688
OG
BSER
A
218
−1.434
17.523
7.675
0.50
19.05
O


ATOM
1689
N
ARG
A
219
−5.054
17.740
7.415
1.00
9.30
N


ATOM
1690
CA
ARG
A
219
−6.019
17.783
8.511
1.00
11.05
C


ATOM
1691
C
ARG
A
219
−7.045
16.657
8.422
1.00
10.10
C


ATOM
1692
O
ARG
A
219
−7.451
16.091
9.439
1.00
10.71
O


ATOM
1693
CB
ARG
A
219
−6.700
19.152
8.579
1.00
10.50
C


ATOM
1694
CG
ARG
A
219
−5.708
20.260
8.925
1.00
10.95
C


ATOM
1695
CD
ARG
A
219
−6.290
21.639
8.732
1.00
13.38
C


ATOM
1696
NE
ARG
A
219
−5.238
22.651
8.755
1.00
13.54
N


ATOM
1697
CZ
ARG
A
219
−5.455
23.955
8.635
1.00
12.88
C


ATOM
1698
NH1
ARG
A
219
−6.690
24.413
8.499
1.00
12.45
N


ATOM
1699
NH2
ARG
A
219
−4.427
24.797
8.662
1.00
14.24
N


ATOM
1700
N
VAL
A
220
−7.466
16.320
7.206
1.00
9.48
N


ATOM
1701
CA
VAL
A
220
−8.391
15.216
7.021
1.00
10.61
C


ATOM
1702
C
VAL
A
220
−7.758
13.921
7.526
1.00
10.79
C


ATOM
1703
O
VAL
A
220
−8.403
13.143
8.223
1.00
11.64
O


ATOM
1704
CB
VAL
A
220
−8.834
15.084
5.547
1.00
9.99
C


ATOM
1705
CG1
VAL
A
220
−9.476
13.727
5.296
1.00
10.95
C


ATOM
1706
CG2
VAL
A
220
−9.802
16.210
5.190
1.00
11.57
C


ATOM
1707
N
GLU
A
221
−6.489
13.706
7.198
1.00
13.35
N


ATOM
1708
CA
GLU
A
221
−5.800
12.492
7.633
1.00
14.85
C


ATOM
1709
C
GLU
A
221
−5.815
12.368
9.157
1.00
18.26
C


ATOM
1710
O
GLU
A
221
−6.082
11.295
9.701
1.00
18.73
O


ATOM
1711
CB
GLU
A
221
−4.362
12.481
7.115
1.00
18.69
C


ATOM
1712
CG
GLU
A
221
−3.543
11.285
7.574
1.00
51.12
C


ATOM
1713
CD
GLU
A
221
−2.103
11.354
7.102
1.00
92.75
C


ATOM
1714
OE1
GLU
A
221
−1.704
12.403
6.551
1.00
79.10
O


ATOM
1715
OE2
GLU
A
221
−1.369
10.358
7.283
1.00
103.53
O


ATOM
1716
N
ARG
A
222
−5.545
13.473
9.844
1.00
14.30
N


ATOM
1717
CA
ARG
A
222
−5.568
13.479
11.305
1.00
15.32
C


ATOM
1718
C
ARG
A
222
−6.967
13.282
11.865
1.00
13.43
C


ATOM
1719
O
ARG
A
222
−7.139
12.698
12.936
1.00
19.93
O


ATOM
1720
CB
ARG
A
222
−4.954
14.774
11.853
1.00
20.09
C


ATOM
1721
CG
ARG
A
222
−3.440
14.809
11.778
1.00
30.84
C


ATOM
1722
CD
ARG
A
222
−2.868
16.013
12.513
1.00
36.59
C


ATOM
1723
NE
ARG
A
222
−3.115
17.262
11.798
1.00
51.40
N


ATOM
1724
CZ
ARG
A
222
−2.332
17.740
10.837
1.00
65.64
C


ATOM
1725
NH1
ARG
A
222
−1.245
17.073
10.471
1.00
72.52
N


ATOM
1726
NH2
ARG
A
222
−2.635
18.885
10.240
1.00
34.72
N


ATOM
1727
N
ALA
A
223
−7.971
13.770
11.149
1.00
12.61
N


ATOM
1728
CA
ALA
A
223
−9.345
13.670
11.607
1.00
14.25
C


ATOM
1729
C
ALA
A
223
−9.941
12.264
11.444
1.00
14.35
C


ATOM
1730
O
ALA
A
223
−10.856
11.894
12.178
1.00
17.85
O


ATOM
1731
CB
ALA
A
223
−10.211
14.692
10.886
1.00
15.85
C


ATOM
1732
N
LEU
A
224
−9.429
11.502
10.476
1.00
14.85
N


ATOM
1733
CA
LEU
A
224
−10.055
10.231
10.085
1.00
19.59
C


ATOM
1734
C
LEU
A
224
−10.274
9.268
11.252
1.00
18.08
C


ATOM
1735
O
LEU
A
224
−11.392
8.799
11.455
1.00
21.08
O


ATOM
1736
CB
LEU
A
224
−9.277
9.537
8.958
1.00
18.13
C


ATOM
1737
CG
LEU
A
224
−9.303
10.188
7.572
1.00
21.40
C


ATOM
1738
CD1
LEU
A
224
−8.687
9.260
6.531
1.00
28.66
C


ATOM
1739
CD2
LEU
A
224
−10.712
10.584
7.173
1.00
23.13
C


ATOM
1740
N
PRO
A
225
−9.213
8.985
12.031
1.00
23.78
N


ATOM
1741
CA
PRO
A
225
−9.334
8.027
13.138
1.00
24.17
C


ATOM
1742
C
PRO
A
225
−10.310
8.479
14.215
1.00
26.85
C


ATOM
1743
O
PRO
A
225
−10.636
7.694
15.106
1.00
34.94
O


ATOM
1744
CB
PRO
A
225
−7.915
7.974
13.713
1.00
28.13
C


ATOM
1745
CG
PRO
A
225
−7.034
8.443
12.608
1.00
36.69
C


ATOM
1746
CD
PRO
A
225
−7.833
9.477
11.884
1.00
20.63
C


ATOM
1747
N
LYS
A
226
−10.771
9.723
14.130
1.00
25.36
N


ATOM
1748
CA
LYS
A
226
−11.684
10.280
15.120
1.00
25.72
C


ATOM
1749
C
LYS
A
226
−13.132
10.300
14.630
1.00
33.01
C


ATOM
1750
O
LYS
A
226
−14.058
10.524
15.408
1.00
28.49
O


ATOM
1751
CB
LYS
A
226
−11.248
11.700
15.486
1.00
44.44
C


ATOM
1752
CG
LYS
A
226
−9.761
11.834
15.776
1.00
42.51
C


ATOM
1753
CD
LYS
A
226
−9.371
13.289
15.988
1.00
69.34
C


ATOM
1754
CE
LYS
A
226
−7.876
13.432
16.224
1.00
70.90
C


ATOM
1755
NZ
LYS
A
226
−7.485
14.853
16.441
1.00
84.95
N


ATOM
1756
N
LEU
A
227
−13.318
10.062
13.336
1.00
20.18
N


ATOM
1757
CA
ALEU
A
227
−14.637
10.116
12.716
0.50
15.98
C


ATOM
1758
C
LEU
A
227
−15.525
8.981
13.183
1.00
13.40
C


ATOM
1759
O
LEU
A
227
−15.109
7.818
13.166
1.00
17.01
O


ATOM
1760
CB
ALEU
A
227
−14.512
10.017
11.196
0.50
17.93
C


ATOM
1761
CG
ALEU
A
227
−14.149
11.259
10.394
0.50
13.32
C


ATOM
1762
CD1
ALEU
A
227
−14.050
10.874
8.923
0.50
13.43
C


ATOM
1763
CD2
ALEU
A
227
−15.179
12.355
10.601
0.50
15.66
C


ATOM
1764
CA
BLEU
A
227
−14.632
10.132
12.717
0.50
15.66
C


ATOM
1765
CB
BLEU
A
227
−14.470
10.100
11.192
0.50
18.07
C


ATOM
1766
CG
BLEU
A
227
−15.573
10.629
10.274
0.50
15.22
C


ATOM
1767
CD1
BLEU
A
227
−15.891
12.088
10.568
0.50
24.87
C


ATOM
1768
CD2
BLEU
A
227
−15.155
10.453
8.822
0.50
11.89
C


ATOM
1769
N
THR
A
228
−16.746
9.311
13.596
1.00
10.05
N


ATOM
1770
CA
THR
A
228
−17.727
8.299
13.997
1.00
12.36
C


ATOM
1771
C
THR
A
228
−19.067
8.438
13.272
1.00
10.02
C


ATOM
1772
O
THR
A
228
−19.946
7.580
13.400
1.00
10.20
O


ATOM
1773
CB
THR
A
228
−17.994
8.324
15.523
1.00
15.46
C


ATOM
1774
OG1
THR
A
228
−18.544
9.596
15.893
1.00
17.33
O


ATOM
1775
CG2
THR
A
228
−16.710
8.077
16.300
1.00
16.53
C


ATOM
1776
N
VAL
A
229
−19.231
9.526
12.518
1.00
10.47
N


ATOM
1777
CA
VAL
A
229
−20.485
9.808
11.826
1.00
9.15
C


ATOM
1778
C
VAL
A
229
−20.779
8.746
10.744
1.00
7.39
C


ATOM
1779
O
VAL
A
229
−19.859
8.294
10.059
1.00
8.94
O


ATOM
1780
CB
VAL
A
229
−20.438
11.233
11.185
1.00
11.08
C


ATOM
1781
CG1
VAL
A
229
−19.367
11.300
10.106
1.00
13.13
C


ATOM
1782
CG2
VAL
A
229
−21.788
11.640
10.626
1.00
12.72
C


ATOM
1783
N
PRO
A
230
−22.041
8.320
10.620
1.00
7.45
N


ATOM
1784
CA
PRO
A
230
−22.387
7.423
9.507
1.00
6.36
C


ATOM
1785
C
PRO
A
230
−22.051
8.070
8.168
1.00
9.18
C


ATOM
1786
O
PRO
A
230
−22.254
9.273
8.002
1.00
10.77
O


ATOM
1787
CB
PRO
A
230
−23.898
7.272
9.630
1.00
9.15
C


ATOM
1788
CG
PRO
A
230
−24.173
7.499
11.112
1.00
9.93
C


ATOM
1789
CD
PRO
A
230
−23.181
8.547
11.527
1.00
10.30
C


ATOM
1790
N
PHE
A
231
−21.543
7.296
7.211
1.00
6.79
N


ATOM
1791
CA
PHE
A
231
−21.343
7.874
5.886
1.00
7.46
C


ATOM
1792
C
PHE
A
231
−21.514
6.920
4.722
1.00
7.73
C


ATOM
1793
O
PHE
A
231
−21.356
5.690
4.853
1.00
7.65
O


ATOM
1794
CB
PHE
A
231
−20.019
8.643
5.779
1.00
7.69
C


ATOM
1795
CG
PHE
A
231
−18.787
7.796
5.905
1.00
7.77
C


ATOM
1796
CD1
PHE
A
231
−18.230
7.177
4.784
1.00
9.21
C


ATOM
1797
CD2
PHE
A
231
−18.145
7.656
7.125
1.00
8.70
C


ATOM
1798
CE1
PHE
A
231
−17.076
6.414
4.892
1.00
10.86
C


ATOM
1799
CE2
PHE
A
231
−16.986
6.901
7.243
1.00
9.68
C


ATOM
1800
CZ
PHE
A
231
−16.446
6.274
6.122
1.00
10.56
C


ATOM
1801
N
LEU
A
232
−21.861
7.532
3.590
1.00
7.02
N


ATOM
1802
CA
LEU
A
232
−21.907
6.871
2.291
1.00
5.67
C


ATOM
1803
C
LEU
A
232
−20.836
7.551
1.462
1.00
7.56
C


ATOM
1804
O
LEU
A
232
−20.824
8.788
1.349
1.00
8.41
O


ATOM
1805
CB
LEU
A
232
−23.257
7.105
1.638
1.00
7.44
C


ATOM
1806
CG
LEU
A
232
−23.360
6.631
0.181
1.00
8.89
C


ATOM
1807
CD1
LEU
A
232
−23.158
5.113
0.097
1.00
9.59
C


ATOM
1808
CD2
LEU
A
232
−24.689
7.035
−0.430
1.00
9.38
C


ATOM
1809
N
LEU
A
233
−19.937
6.755
0.893
1.00
5.98
N


ATOM
1810
CA
LEU
A
233
−18.816
7.251
0.123
1.00
5.46
C


ATOM
1811
C
LEU
A
233
−18.961
6.736
−1.311
1.00
6.46
C


ATOM
1812
O
LEU
A
233
−19.020
5.524
−1.546
1.00
8.99
O


ATOM
1813
CB
LEU
A
233
−17.536
6.719
0.749
1.00
6.96
C


ATOM
1814
CG
LEU
A
233
−16.204
6.995
0.085
1.00
11.73
C


ATOM
1815
CD1
LEU
A
233
−15.985
8.484
−0.098
1.00
11.86
C


ATOM
1816
CD2
LEU
A
233
−15.087
6.376
0.926
1.00
11.71
C


ATOM
1817
N
LEU
A
234
−19.047
7.664
−2.266
1.00
6.30
N


ATOM
1818
CA
LEU
A
234
−19.146
7.328
−3.690
1.00
5.53
C


ATOM
1819
C
LEU
A
234
−17.887
7.806
−4.387
1.00
6.67
C


ATOM
1820
O
LEU
A
234
−17.482
8.964
−4.214
1.00
7.50
O


ATOM
1821
CB
LEU
A
234
−20.366
8.015
−4.299
1.00
7.09
C


ATOM
1822
CG
LEU
A
234
−21.692
7.778
−3.587
1.00
7.45
C


ATOM
1823
CD1
LEU
A
234
−22.809
8.556
−4.278
1.00
12.21
C


ATOM
1824
CD2
LEU
A
234
−22.056
6.295
−3.498
1.00
10.51
C


ATOM
1825
N
GLN
A
235
−17.263
6.930
−5.172
1.00
6.02
N


ATOM
1826
CA
GLN
A
235
−15.962
7.230
−5.746
1.00
6.69
C


ATOM
1827
C
GLN
A
235
−15.826
6.601
−7.122
1.00
7.26
C


ATOM
1828
O
GLN
A
235
−16.188
5.433
−7.310
1.00
9.83
O


ATOM
1829
CB
GLN
A
235
−14.878
6.664
−4.823
1.00
8.10
C


ATOM
1830
CG
GLN
A
235
−13.460
6.805
−5.331
1.00
8.78
C


ATOM
1831
CD
GLN
A
235
−12.945
8.235
−5.263
1.00
14.60
C


ATOM
1832
OE1
GLN
A
235
−13.229
8.969
−4.310
1.00
11.92
O


ATOM
1833
NE2
GLN
A
235
−12.176
8.639
−6.273
1.00
14.30
N


ATOM
1834
N
GLY
A
236
−15.324
7.367
−8.092
1.00
7.16
N


ATOM
1835
CA
GLY
A
236
−15.027
6.814
−9.410
1.00
7.05
C


ATOM
1836
C
GLY
A
236
−13.626
6.241
−9.448
1.00
7.13
C


ATOM
1837
O
GLY
A
236
−12.698
6.781
−8.847
1.00
9.31
O


ATOM
1838
N
SER
A
237
−13.451
5.143
−10.182
1.00
7.90
N


ATOM
1839
CA
SER
A
237
−12.139
4.515
−10.244
1.00
10.56
C


ATOM
1840
C
SER
A
237
−11.158
5.288
−11.122
1.00
8.69
C


ATOM
1841
O
SER
A
237
−9.947
5.107
−11.016
1.00
12.81
O


ATOM
1842
CB
SER
A
237
−12.244
3.065
−10.734
1.00
14.27
C


ATOM
1843
OG
SER
A
237
−12.468
3.007
−12.137
1.00
14.14
O


ATOM
1844
N
ALA
A
238
−11.676
6.132
−12.009
1.00
8.64
N


ATOM
1845
CA
ALA
A
238
−10.806
6.884
−12.918
1.00
12.93
C


ATOM
1846
C
ALA
A
238
−10.783
8.358
−12.539
1.00
12.31
C


ATOM
1847
O
ALA
A
238
−10.740
9.239
−13.402
1.00
11.93
O


ATOM
1848
CB
ALA
A
238
−11.257
6.704
−14.359
1.00
11.58
C


ATOM
1849
N
ASP
A
239
−10.803
8.616
−11.238
1.00
9.91
N


ATOM
1850
CA
ASP
A
239
−10.753
9.975
−10.703
1.00
11.61
C


ATOM
1851
C
ASP
A
239
−9.331
10.528
−10.745
1.00
11.01
C


ATOM
1852
O
ASP
A
239
−8.434
10.018
−10.070
1.00
12.82
O


ATOM
1853
CB
ASP
A
239
−11.255
9.950
−9.260
1.00
9.49
C


ATOM
1854
CG
ASP
A
239
−11.495
11.338
−8.701
1.00
10.07
C


ATOM
1855
OD1
ASP
A
239
−10.756
12.276
−9.080
1.00
9.43
O


ATOM
1856
OD2
ASP
A
239
−12.430
11.490
−7.888
1.00
10.75
O


ATOM
1857
N
ARG
A
240
−9.131
11.573
−11.549
1.00
10.38
N


ATOM
1858
CA
ARG
A
240
−7.807
12.154
−11.748
1.00
11.86
C


ATOM
1859
C
ARG
A
240
−7.387
13.173
−10.695
1.00
13.94
C


ATOM
1860
O
ARG
A
240
−6.243
13.631
−10.692
1.00
15.63
O


ATOM
1861
CB
ARG
A
240
−7.736
12.819
−13.124
1.00
15.76
C


ATOM
1862
CG
ARG
A
240
−7.703
11.848
−14.277
1.00
26.43
C


ATOM
1863
CD
ARG
A
240
−6.677
12.317
−15.281
1.00
32.76
C


ATOM
1864
NE
ARG
A
240
−6.988
11.882
−16.633
1.00
71.10
N


ATOM
1865
CZ
ARG
A
240
−6.466
12.441
−17.716
1.00
19.70
C


ATOM
1866
NH1
ARG
A
240
−5.618
13.457
−17.587
1.00
26.60
N


ATOM
1867
NH2
ARG
A
240
−6.798
11.989
−18.914
1.00
79.96
N


ATOM
1868
N
LEU
A
241
−8.305
13.521
−9.803
1.00
10.36
N


ATOM
1869
CA
LEU
A
241
−8.077
14.586
−8.841
1.00
12.06
C


ATOM
1870
C
LEU
A
241
−8.030
14.068
−7.404
1.00
14.79
C


ATOM
1871
O
LEU
A
241
−7.048
14.283
−6.685
1.00
17.27
O


ATOM
1872
CB
LEU
A
241
−9.189
15.621
−8.995
1.00
12.48
C


ATOM
1873
CG
LEU
A
241
−8.962
16.968
−8.329
1.00
17.56
C


ATOM
1874
CD1
LEU
A
241
−7.606
17.547
−8.721
1.00
17.75
C


ATOM
1875
CD2
LEU
A
241
−10.086
17.899
−8.744
1.00
14.80
C


ATOM
1876
N
CYS
A
242
−9.103
13.402
−6.990
1.00
13.17
N


ATOM
1877
CA
CYS
A
242
−9.172
12.777
−5.674
1.00
14.00
C


ATOM
1878
C
CYS
A
242
−8.910
11.295
−5.861
1.00
12.30
C


ATOM
1879
O
CYS
A
242
−9.775
10.554
−6.343
1.00
16.19
O


ATOM
1880
CB
CYS
A
242
−10.531
13.035
−5.026
1.00
14.14
C


ATOM
1881
SG
CYS
A
242
−10.791
14.786
−4.632
1.00
17.18
S


ATOM
1882
N
ASP
A
243
−7.701
10.872
−5.514
1.00
14.74
N


ATOM
1883
CA
ASP
A
243
−7.270
9.511
−5.787
1.00
15.90
C


ATOM
1884
C
ASP
A
243
−8.125
8.489
−5.046
1.00
18.69
C


ATOM
1885
O
ASP
A
243
−8.421
8.649
−3.860
1.00
14.40
O


ATOM
1886
CB
ASP
A
243
−5.804
9.327
−5.418
1.00
17.66
C


ATOM
1887
CG
ASP
A
243
−5.180
8.144
−6.122
1.00
49.96
C


ATOM
1888
OD1
ASP
A
243
−4.757
8.299
−7.289
1.00
39.78
O


ATOM
1889
OD2
ASP
A
243
−5.123
7.057
−5.512
1.00
25.88
O


ATOM
1890
N
SER
A
244
−8.527
7.436
−5.748
1.00
15.66
N


ATOM
1891
CA
SER
A
244
−9.378
6.416
−5.142
1.00
17.72
C


ATOM
1892
C
SER
A
244
−8.736
5.808
−3.894
1.00
15.64
C


ATOM
1893
O
SER
A
244
−9.435
5.302
−3.025
1.00
15.29
O


ATOM
1894
CB
SER
A
244
−9.721
5.322
−6.155
1.00
17.74
C


ATOM
1895
OG
SER
A
244
−8.542
4.671
−6.580
1.00
20.33
O


ATOM
1896
N
LYS
A
245
−7.412
5.860
−3.800
1.00
14.32
N


ATOM
1897
CA
LYS
A
245
−6.711
5.394
−2.606
1.00
12.72
C


ATOM
1898
C
LYS
A
245
−7.198
6.097
−1.339
1.00
16.08
C


ATOM
1899
O
LYS
A
245
−7.211
5.508
−0.255
1.00
15.31
O


ATOM
1900
CB
LYS
A
245
−5.203
5.593
−2.754
1.00
18.83
C


ATOM
1901
CG
LYS
A
245
−4.394
5.124
−1.557
1.00
44.09
C


ATOM
1902
CD
LYS
A
245
−2.929
5.507
−1.692
1.00
75.26
C


ATOM
1903
CE
LYS
A
245
−2.150
5.169
−0.430
1.00
60.92
C


ATOM
1904
NZ
LYS
A
245
−0.732
5.618
−0.513
1.00
81.77
N


ATOM
1905
N
GLY
A
246
−7.597
7.357
−1.475
1.00
12.05
N


ATOM
1906
CA
GLY
A
246
−8.129
8.112
−0.354
1.00
11.97
C


ATOM
1907
C
GLY
A
246
−9.462
7.569
0.115
1.00
12.09
C


ATOM
1908
O
GLY
A
246
−9.752
7.562
1.312
1.00
11.82
O


ATOM
1909
N
ALA
A
247
−10.288
7.118
−0.820
1.00
11.78
N


ATOM
1910
CA
ALA
A
247
−11.562
6.515
−0.461
1.00
12.42
C


ATOM
1911
C
ALA
A
247
−11.315
5.279
0.404
1.00
11.27
C


ATOM
1912
O
ALA
A
247
−11.993
5.079
1.417
1.00
11.32
O


ATOM
1913
CB
ALA
A
247
−12.369
6.151
−1.713
1.00
11.97
C


ATOM
1914
N
TYR
A
248
−10.349
4.451
0.001
1.00
11.26
N


ATOM
1915
CA
TYR
A
248
−10.022
3.256
0.776
1.00
12.36
C


ATOM
1916
C
TYR
A
248
−9.458
3.631
2.155
1.00
12.09
C


ATOM
1917
O
TYR
A
248
−9.752
2.971
3.152
1.00
11.59
O


ATOM
1918
CB
TYR
A
248
−9.074
2.322
0.001
1.00
11.60
C


ATOM
1919
CG
TYR
A
248
−9.652
1.861
−1.322
1.00
14.61
C


ATOM
1920
CD1
TYR
A
248
−10.872
1.201
−1.371
1.00
13.55
C


ATOM
1921
CD2
TYR
A
248
−8.985
2.092
−2.520
1.00
19.12
C


ATOM
1922
CE1
TYR
A
248
−11.419
0.784
−2.571
1.00
18.03
C


ATOM
1923
CE2
TYR
A
248
−9.528
1.677
−3.728
1.00
19.37
C


ATOM
1924
CZ
TYR
A
248
−10.743
1.025
−3.742
1.00
16.03
C


ATOM
1925
OH
TYR
A
248
−11.302
0.600
−4.929
1.00
19.87
O


ATOM
1926
N
LEU
A
249
−8.673
4.703
2.215
1.00
11.41
N


ATOM
1927
CA
LEU
A
249
−8.116
5.181
3.484
1.00
10.25
C


ATOM
1928
C
LEU
A
249
−9.220
5.627
4.447
1.00
10.35
C


ATOM
1929
O
LEU
A
249
−9.189
5.328
5.647
1.00
10.53
O


ATOM
1930
CB
LEU
A
249
−7.150
6.339
3.226
1.00
13.46
C


ATOM
1931
CG
LEU
A
249
−6.365
6.824
4.444
1.00
18.08
C


ATOM
1932
CD1
LEU
A
249
−5.351
5.760
4.858
1.00
19.06
C


ATOM
1933
CD2
LEU
A
249
−5.667
8.146
4.150
1.00
18.48
C


ATOM
1934
N
LEU
A
250
−10.209
6.336
3.923
1.00
9.37
N


ATOM
1935
CA
LEU
A
250
−11.329
6.757
4.724
1.00
9.21
C


ATOM
1936
C
LEU
A
250
−12.080
5.543
5.271
1.00
12.66
C


ATOM
1937
O
LEU
A
250
−12.435
5.510
6.450
1.00
11.83
O


ATOM
1938
CB
LEU
A
250
−12.235
7.661
3.897
1.00
13.06
C


ATOM
1939
CG
LEU
A
250
−13.559
8.092
4.496
1.00
13.37
C


ATOM
1940
CD1
LEU
A
250
−13.375
8.747
5.858
1.00
14.48
C


ATOM
1941
CD2
LEU
A
250
−14.209
9.066
3.529
1.00
16.04
C


ATOM
1942
N
MET
A
251
−12.304
4.531
4.431
1.00
9.91
N


ATOM
1943
CA
MET
A
251
−12.998
3.327
4.888
1.00
10.56
C


ATOM
1944
C
MET
A
251
−12.189
2.596
5.950
1.00
10.04
C


ATOM
1945
O
MET
A
251
−12.759
1.991
6.860
1.00
13.87
O


ATOM
1946
CB
MET
A
251
−13.284
2.381
3.719
1.00
9.50
C


ATOM
1947
CG
MET
A
251
−14.342
2.899
2.762
1.00
13.01
C


ATOM
1948
SD
MET
A
251
−15.992
3.057
3.473
1.00
11.08
S


ATOM
1949
CE
MET
A
251
−16.458
1.331
3.656
1.00
12.29
C


ATOM
1950
N
GLU
A
252
−10.872
2.640
5.835
1.00
8.28
N


ATOM
1951
CA
GLU
A
252
−10.013
1.916
6.763
1.00
10.79
C


ATOM
1952
C
GLU
A
252
−9.890
2.619
8.110
1.00
14.17
C


ATOM
1953
O
GLU
A
252
−9.960
1.979
9.162
1.00
16.34
O


ATOM
1954
CB
GLU
A
252
−8.620
1.717
6.171
1.00
14.04
C


ATOM
1955
CG
GLU
A
252
−7.682
0.962
7.109
1.00
18.86
C


ATOM
1956
CD
GLU
A
252
−6.248
0.940
6.620
1.00
88.82
C


ATOM
1957
OE1
GLU
A
252
−5.970
1.541
5.561
1.00
68.03
O


ATOM
1958
OE2
GLU
A
252
−5.398
0.324
7.297
1.00
99.02
O


ATOM
1959
N
LEU
A
253
−9.701
3.933
8.082
1.00
11.41
N


ATOM
1960
CA
LEU
A
253
−9.336
4.662
9.294
1.00
11.04
C


ATOM
1961
C
LEU
A
253
−10.506
5.218
10.100
1.00
11.58
C


ATOM
1962
O
LEU
A
253
−10.383
5.395
11.309
1.00
14.92
O


ATOM
1963
CB
LEU
A
253
−8.355
5.785
8.958
1.00
14.04
C


ATOM
1964
CG
LEU
A
253
−6.961
5.350
8.512
1.00
15.41
C


ATOM
1965
CD1
LEU
A
253
−6.066
6.565
8.335
1.00
19.75
C


ATOM
1966
CD2
LEU
A
253
−6.354
4.367
9.504
1.00
26.28
C


ATOM
1967
N
ALA
A
254
−11.625
5.526
9.451
1.00
9.74
N


ATOM
1968
CA
ALA
A
254
−12.783
6.055
10.171
1.00
10.79
C


ATOM
1969
C
ALA
A
254
−13.301
5.031
11.184
1.00
12.72
C


ATOM
1970
O
ALA
A
254
−13.301
3.828
10.913
1.00
13.12
O


ATOM
1971
CB
ALA
A
254
−13.887
6.452
9.198
1.00
13.02
C


ATOM
1972
N
LYS
A
255
−13.738
5.502
12.347
1.00
11.30
N


ATOM
1973
CA
LYS
A
255
−14.265
4.593
13.360
1.00
13.33
C


ATOM
1974
C
LYS
A
255
−15.774
4.394
13.233
1.00
14.71
C


ATOM
1975
O
LYS
A
255
−16.377
3.634
13.992
1.00
16.28
O


ATOM
1976
CB
LYS
A
255
−13.899
5.072
14.772
1.00
17.14
C


ATOM
1977
CG
LYS
A
255
−12.399
5.098
15.036
1.00
23.75
C


ATOM
1978
CD
LYS
A
255
−11.776
3.721
14.861
1.00
30.83
C


ATOM
1979
CE
LYS
A
255
−10.267
3.744
15.092
1.00
45.25
C


ATOM
1980
NZ
LYS
A
255
−9.509
4.223
13.898
1.00
43.77
N


ATOM
1981
N
SER
A
256
−16.387
5.067
12.265
1.00
10.29
N


ATOM
1982
CA
SER
A
256
−17.821
4.987
12.074
1.00
9.83
C


ATOM
1983
C
SER
A
256
−18.307
3.548
12.017
1.00
9.52
C


ATOM
1984
O
SER
A
256
−17.734
2.720
11.306
1.00
12.49
O


ATOM
1985
CB
SER
A
256
−18.221
5.704
10.779
1.00
11.18
C


ATOM
1986
OG
SER
A
256
−17.631
6.993
10.743
1.00
11.20
O


ATOM
1987
N
GLN
A
257
−19.374
3.264
12.747
1.00
9.12
N


ATOM
1988
CA
GLN
A
257
−20.003
1.942
12.704
1.00
12.22
C


ATOM
1989
C
GLN
A
257
−20.865
1.739
11.464
1.00
13.06
C


ATOM
1990
O
GLN
A
257
−21.242
0.613
11.143
1.00
14.98
O


ATOM
1991
CB
GLN
A
257
−20.847
1.714
13.959
1.00
13.19
C


ATOM
1992
CG
GLN
A
257
−20.018
1.658
15.238
1.00
16.99
C


ATOM
1993
CD
GLN
A
257
−20.834
1.258
16.450
1.00
67.11
C


ATOM
1994
OE1
GLN
A
257
−22.052
1.432
16.479
1.00
74.68
O


ATOM
1995
NE2
GLN
A
257
−20.163
0.719
17.462
1.00
83.33
N


ATOM
1996
N
ASP
A
258
−21.186
2.829
10.772
1.00
8.32
N


ATOM
1997
CA
ASP
A
258
−22.065
2.767
9.614
1.00
9.96
C


ATOM
1998
C
ASP
A
258
−21.342
3.413
8.433
1.00
9.10
C


ATOM
1999
O
ASP
A
258
−21.434
4.625
8.226
1.00
10.80
O


ATOM
2000
CB
ASP
A
258
−23.365
3.512
9.942
1.00
8.32
C


ATOM
2001
CG
ASP
A
258
−24.395
3.468
8.825
1.00
11.84
C


ATOM
2002
OD1
ASP
A
258
−24.184
2.773
7.806
1.00
11.26
O


ATOM
2003
OD2
ASP
A
258
−25.450
4.120
8.973
1.00
13.37
O


ATOM
2004
N
LYS
A
259
−20.599
2.614
7.677
1.00
6.95
N


ATOM
2005
CA
LYS
A
259
−19.839
3.150
6.547
1.00
7.68
C


ATOM
2006
C
LYS
A
259
−19.924
2.230
5.339
1.00
9.90
C


ATOM
2007
O
LYS
A
259
−19.757
1.001
5.443
1.00
10.58
O


ATOM
2008
CB
LYS
A
259
−18.389
3.463
6.914
1.00
12.50
C


ATOM
2009
CG
LYS
A
259
−17.667
2.376
7.675
1.00
10.44
C


ATOM
2010
CD
LYS
A
259
−16.259
2.799
8.055
1.00
10.64
C


ATOM
2011
CE
LYS
A
259
−15.610
1.749
8.959
1.00
11.27
C


ATOM
2012
NZ
LYS
A
259
−14.164
1.975
9.132
1.00
11.78
N


ATOM
2013
N
THR
A
260
−20.192
2.846
4.192
1.00
6.48
N


ATOM
2014
CA
ATHR
A
260
−20.426
2.128
2.937
0.70
5.21
C


ATOM
2015
C
THR
A
260
−19.679
2.820
1.804
1.00
7.56
C


ATOM
2016
O
THR
A
260
−19.699
4.058
1.710
1.00
8.70
O


ATOM
2017
CB
ATHR
A
260
−21.925
2.119
2.590
0.70
7.13
C


ATOM
2018
OG1
ATHR
A
260
−22.686
1.596
3.693
0.70
8.97
O


ATOM
2019
CG2
ATHR
A
260
−22.190
1.270
1.340
0.70
8.35
C


ATOM
2020
CA
BTHR
A
260
−20.360
2.105
2.955
0.30
8.55
C


ATOM
2021
CB
BTHR
A
260
−21.842
1.896
2.629
0.30
10.28
C


ATOM
2022
OG1
BTHR
A
260
−22.498
3.164
2.524
0.30
15.39
O


ATOM
2023
CG2
BTHR
A
260
−22.511
1.083
3.722
0.30
17.62
C


ATOM
2024
N
LEU
A
261
−19.051
2.029
0.942
1.00
5.66
N


ATOM
2025
CA
LEU
A
261
−18.343
2.540
−0.225
1.00
7.03
C


ATOM
2026
C
LEU
A
261
−18.930
1.930
−1.486
1.00
8.11
C


ATOM
2027
O
LEU
A
261
−19.079
0.710
−1.580
1.00
8.85
O


ATOM
2028
CB
LEU
A
261
−16.878
2.162
−0.147
1.00
8.55
C


ATOM
2029
CG
LEU
A
261
−16.055
2.355
−1.429
1.00
7.61
C


ATOM
2030
CD1
LEU
A
261
−15.940
3.844
−1.802
1.00
10.12
C


ATOM
2031
CD2
LEU
A
261
−14.679
1.753
−1.241
1.00
9.79
C


ATOM
2032
N
LYS
A
262
−19.273
2.775
−2.459
1.00
5.57
N


ATOM
2033
CA
LYS
A
262
−19.587
2.288
−3.810
1.00
7.37
C


ATOM
2034
C
LYS
A
262
−18.569
2.856
−4.785
1.00
9.49
C


ATOM
2035
O
LYS
A
262
−18.314
4.066
−4.798
1.00
9.23
O


ATOM
2036
CB
LYS
A
262
−20.996
2.686
−4.224
1.00
9.44
C


ATOM
2037
CG
LYS
A
262
−21.487
2.018
−5.527
1.00
14.01
C


ATOM
2038
CD
LYS
A
262
−22.988
2.224
−5.735
1.00
17.20
C


ATOM
2039
CE
LYS
A
262
−23.449
1.716
−7.104
1.00
18.72
C


ATOM
2040
NZ
LYS
A
262
−23.405
0.223
−7.191
1.00
18.42
N


ATOM
2041
N
ILE
A
263
−17.949
1.975
−5.567
1.00
7.42
N


ATOM
2042
CA
AILE
A
263
−16.971
2.372
−6.583
0.90
9.22
C


ATOM
2043
C
ILE
A
263
−17.617
2.289
−7.953
1.00
7.84
C


ATOM
2044
O
ILE
A
263
−18.215
1.266
−8.303
1.00
9.35
O


ATOM
2045
CB
AILE
A
263
−15.715
1.470
−6.574
0.90
9.09
C


ATOM
2046
CG1
AILE
A
263
−14.885
1.681
−5.301
0.90
11.72
C


ATOM
2047
CG2
AILE
A
263
−14.865
1.719
−7.834
0.90
11.99
C


ATOM
2048
CD1
AILE
A
263
−14.085
2.991
−5.280
0.90
11.94
C


ATOM
2049
CA
BILE
A
263
−16.994
2.399
−6.575
0.10
10.98
C


ATOM
2050
CB
BILE
A
263
−15.721
1.559
−6.521
0.10
6.66
C


ATOM
2051
CG1
BILE
A
263
−15.124
1.623
−5.119
0.10
7.98
C


ATOM
2052
CG2
BILE
A
263
−14.718
2.040
−7.561
0.10
17.27
C


ATOM
2053
CD1
BILE
A
263
−13.981
0.698
−4.927
0.10
3.99
C


ATOM
2054
N
TYR
A
264
−17.498
3.363
−8.726
1.00
7.78
N


ATOM
2055
CA
TYR
A
264
−18.012
3.401
−10.095
1.00
9.35
C


ATOM
2056
C
TYR
A
264
−16.856
3.194
−11.063
1.00
8.73
C


ATOM
2057
O
TYR
A
264
−16.002
4.063
−11.239
1.00
7.97
O


ATOM
2058
CB
TYR
A
264
−18.727
4.723
−10.348
1.00
9.49
C


ATOM
2059
CG
TYR
A
264
−20.043
4.815
−9.615
1.00
5.92
C


ATOM
2060
CD1
TYR
A
264
−21.221
4.371
−10.195
1.00
9.53
C


ATOM
2061
CD2
TYR
A
264
−20.108
5.343
−8.330
1.00
10.30
C


ATOM
2062
CE1
TYR
A
264
−22.440
4.448
−9.525
1.00
10.75
C


ATOM
2063
CE2
TYR
A
264
−21.316
5.425
−7.650
1.00
8.15
C


ATOM
2064
CZ
TYR
A
264
−22.476
4.978
−8.241
1.00
8.75
C


ATOM
2065
OH
TYR
A
264
−23.684
5.059
−7.585
1.00
10.60
O


ATOM
2066
N
GLU
A
265
−16.811
2.008
−11.667
1.00
9.58
N


ATOM
2067
CA
GLU
A
265
−15.690
1.611
−12.504
1.00
12.10
C


ATOM
2068
C
GLU
A
265
−15.638
2.432
−13.796
1.00
8.46
C


ATOM
2069
O
GLU
A
265
−16.606
2.477
−14.549
1.00
11.89
O


ATOM
2070
CB
GLU
A
265
−15.789
0.114
−12.824
1.00
16.13
C


ATOM
2071
CG
GLU
A
265
−14.547
−0.464
−13.462
1.00
17.85
C


ATOM
2072
CD
GLU
A
265
−13.347
−0.469
−12.534
1.00
21.43
C


ATOM
2073
OE1
GLU
A
265
−13.508
−0.218
−11.312
1.00
20.61
O


ATOM
2074
OE2
GLU
A
265
−12.227
−0.726
−13.020
1.00
32.40
O


ATOM
2075
N
GLY
A
266
−14.507
3.096
−14.005
1.00
9.44
N


ATOM
2076
CA
GLY
A
266
−14.289
3.887
−15.205
1.00
11.21
C


ATOM
2077
C
GLY
A
266
−14.797
5.313
−15.075
1.00
11.31
C


ATOM
2078
O
GLY
A
266
−14.514
6.145
−15.936
1.00
10.95
O


ATOM
2079
N
ALA
A
267
−15.548
5.598
−14.015
1.00
9.68
N


ATOM
2080
CA
ALA
A
267
−16.150
6.920
−13.851
1.00
10.27
C


ATOM
2081
C
ALA
A
267
−15.114
7.942
−13.397
1.00
6.06
C


ATOM
2082
O
ALA
A
267
−14.166
7.627
−12.691
1.00
7.71
O


ATOM
2083
CB
ALA
A
267
−17.324
6.859
−12.875
1.00
9.22
C


ATOM
2084
N
TYR
A
268
−15.291
9.176
−13.865
1.00
7.32
N


ATOM
2085
CA
TYR
A
268
−14.391
10.267
−13.504
1.00
7.50
C


ATOM
2086
C
TYR
A
268
−14.858
10.929
−12.194
1.00
9.01
C


ATOM
2087
O
TYR
A
268
−15.804
10.481
−11.556
1.00
11.16
O


ATOM
2088
CB
TYR
A
268
−14.357
11.317
−14.632
1.00
8.84
C


ATOM
2089
CG
TYR
A
268
−13.629
10.892
−15.905
1.00
9.04
C


ATOM
2090
CD1
TYR
A
268
−13.218
9.575
−16.106
1.00
12.36
C


ATOM
2091
CD2
TYR
A
268
−13.385
11.812
−16.915
1.00
10.68
C


ATOM
2092
CE1
TYR
A
268
−12.557
9.193
−17.278
1.00
13.18
C


ATOM
2093
CE2
TYR
A
268
−12.736
11.442
−18.087
1.00
15.29
C


ATOM
2094
CZ
TYR
A
268
−12.324
10.140
−18.259
1.00
14.67
C


ATOM
2095
OH
TYR
A
268
−11.687
9.783
−19.436
1.00
17.75
O


ATOM
2096
N
HIS
A
269
−14.191
12.013
−11.834
1.00
7.57
N


ATOM
2097
CA
HIS
A
269
−14.368
12.696
−10.548
1.00
6.36
C


ATOM
2098
C
HIS
A
269
−15.795
13.154
−10.224
1.00
7.22
C


ATOM
2099
O
HIS
A
269
−16.295
12.928
−9.115
1.00
7.15
O


ATOM
2100
CB
HIS
A
269
−13.421
13.898
−10.549
1.00
7.11
C


ATOM
2101
CG
HIS
A
269
−13.401
14.689
−9.274
1.00
7.54
C


ATOM
2102
ND1
HIS
A
269
−13.058
14.144
−8.053
1.00
9.09
N


ATOM
2103
CD2
HIS
A
269
−13.599
16.012
−9.051
1.00
9.50
C


ATOM
2104
CE1
HIS
A
269
−13.070
15.096
−7.130
1.00
8.18
C


ATOM
2105
NE2
HIS
A
269
−13.393
16.237
−7.711
1.00
10.12
N


ATOM
2106
N
VAL
A
270
−16.464
13.813
−11.161
1.00
6.45
N


ATOM
2107
CA
VAL
A
270
−17.721
14.471
−10.832
1.00
6.65
C


ATOM
2108
C
VAL
A
270
−18.897
13.531
−11.071
1.00
7.38
C


ATOM
2109
O
VAL
A
270
−19.610
13.613
−12.081
1.00
6.78
O


ATOM
2110
CB
VAL
A
270
−17.896
15.798
−11.602
1.00
5.79
C


ATOM
2111
CG1
VAL
A
270
−19.038
16.584
−10.997
1.00
7.82
C


ATOM
2112
CG2
VAL
A
270
−16.630
16.622
−11.516
1.00
7.71
C


ATOM
2113
N
LEU
A
271
−19.103
12.637
−10.108
1.00
6.65
N


ATOM
2114
CA
LEU
A
271
−20.053
11.534
−10.285
1.00
6.15
C


ATOM
2115
C
LEU
A
271
−21.499
11.970
−10.477
1.00
5.91
C


ATOM
2116
O
LEU
A
271
−22.285
11.283
−11.133
1.00
7.39
O


ATOM
2117
CB
LEU
A
271
−19.957
10.556
−9.105
1.00
6.48
C


ATOM
2118
CG
LEU
A
271
−18.615
9.871
−8.906
1.00
5.62
C


ATOM
2119
CD1
LEU
A
271
−18.659
9.055
−7.613
1.00
9.62
C


ATOM
2120
CD2
LEU
A
271
−18.321
8.957
−10.089
1.00
8.14
C


ATOM
2121
N
HIS
A
272
−21.873
13.103
−9.886
1.00
5.74
N


ATOM
2122
CA
HIS
A
272
−23.233
13.610
−10.006
1.00
5.90
C


ATOM
2123
C
HIS
A
272
−23.448
14.420
−11.291
1.00
5.61
C


ATOM
2124
O
HTS
A
272
−24.538
14.940
−11.511
1.00
8.27
O


ATOM
2125
CB
HIS
A
272
−23.602
14.449
−8.773
1.00
6.29
C


ATOM
2126
CG
HIS
A
272
−22.612
15.529
−8.463
1.00
8.33
C


ATOM
2127
ND1
HIS
A
272
−21.309
15.262
−8.108
1.00
8.50
N


ATOM
2128
CD2
HIS
A
272
−22.731
16.879
−8.481
1.00
11.04
C


ATOM
2129
CE1
HIS
A
272
−20.669
16.402
−7.905
1.00
12.80
C


ATOM
2130
NE2
HIS
A
272
−21.510
17.396
−8.128
1.00
11.63
N


ATOM
2131
N
LYS
A
273
−22.403
14.505
−12.121
1.00
6.91
N


ATOM
2132
CA
LYS
A
273
−22.487
15.162
−13.432
1.00
7.10
C


ATOM
2133
C
LYS
A
273
−21.801
14.293
−14.470
1.00
7.81
C


ATOM
2134
O
LYS
A
273
−21.114
14.800
−15.363
1.00
8.13
O


ATOM
2135
CB
LYS
A
273
−21.778
16.519
−13.399
1.00
8.99
C


ATOM
2136
CG
LYS
A
273
−22.307
17.505
−12.362
1.00
9.49
C


ATOM
2137
CD
LYS
A
273
−23.666
18.011
−12.739
1.00
9.39
C


ATOM
2138
CE
LYS
A
273
−24.085
19.150
−11.798
1.00
11.48
C


ATOM
2139
NZ
LYS
A
273
−25.494
19.505
−12.017
1.00
16.06
N


ATOM
2140
N
GLU
A
274
−21.948
12.978
−14.319
1.00
7.75
N


ATOM
2141
CA
GLU
A
274
−21.254
12.023
−15.169
1.00
7.79
C


ATOM
2142
C
GLU
A
274
−22.186
11.460
−16.249
1.00
7.03
C


ATOM
2143
O
GLU
A
274
−23.236
12.024
−16.542
1.00
9.33
O


ATOM
2144
CB
GLU
A
274
−20.654
10.908
−14.297
1.00
10.21
C


ATOM
2145
CG
GLU
A
274
−19.145
10.870
−14.300
1.00
9.08
C


ATOM
2146
CD
GLU
A
274
−18.598
10.004
−15.429
1.00
8.56
C


ATOM
2147
OE1
GLU
A
274
−17.403
10.118
−15.749
1.00
10.56
O


ATOM
2148
OE2
GLU
A
274
−19.388
9.210
−15.979
1.00
11.18
O


ATOM
2149
N
LEU
A
275
−21.787
10.352
−16.869
1.00
8.50
N


ATOM
2150
CA
LEU
A
275
−22.669
9.703
−17.836
1.00
9.61
C


ATOM
2151
C
LEU
A
275
−23.985
9.365
−17.146
1.00
11.58
C


ATOM
2152
O
LEU
A
275
−23.991
9.062
−15.954
1.00
10.17
O


ATOM
2153
CB
LEU
A
275
−22.016
8.439
−18.394
1.00
9.97
C


ATOM
2154
CG
LEU
A
275
−20.737
8.703
−19.181
1.00
12.18
C


ATOM
2155
CD1
LEU
A
275
−20.108
7.415
−19.713
1.00
14.42
C


ATOM
2156
CD2
LEU
A
275
−21.026
9.667
−20.329
1.00
15.54
C


ATOM
2157
N
PRO
A
276
−25.107
9.413
−17.881
1.00
11.73
N


ATOM
2158
CA
PRO
A
276
−26.427
9.164
−17.288
1.00
14.10
C


ATOM
2159
C
PRO
A
276
−26.511
7.906
−16.417
1.00
16.89
C


ATOM
2160
O
PRO
A
276
−27.109
7.964
−15.336
1.00
17.36
O


ATOM
2161
CB
PRO
A
276
−27.334
9.016
−18.512
1.00
16.90
C


ATOM
2162
CG
PRO
A
276
−26.692
9.878
−19.538
1.00
21.28
C


ATOM
2163
CD
PRO
A
276
−25.205
9.809
−19.300
1.00
13.96
C


ATOM
2164
N
GLU
A
277
−25.939
6.792
−16.870
1.00
14.89
N


ATOM
2165
CA
GLU
A
277
−25.994
5.547
−16.100
1.00
16.63
C


ATOM
2166
C
GLU
A
277
−25.385
5.732
−14.712
1.00
16.38
C


ATOM
2167
O
GLU
A
277
−25.934
5.261
−13.712
1.00
19.95
O


ATOM
2168
CB
GLU
A
277
−25.286
4.408
−16.841
1.00
21.94
C


ATOM
2169
CG
GLU
A
277
−26.111
3.796
−17.961
1.00
85.26
C


ATOM
2170
CD
GLU
A
277
−25.461
2.565
−18.564
1.00
121.84
C


ATOM
2171
OE1
GLU
A
277
−24.359
2.187
−18.112
1.00
109.89
O


ATOM
2172
OE2
GLU
A
277
−26.055
1.973
−19.490
1.00
104.17
O


ATOM
2173
N
VAL
A
278
−24.252
6.421
−14.655
1.00
10.73
N


ATOM
2174
CA
VAL
A
278
−23.595
6.693
−13.384
1.00
9.85
C


ATOM
2175
C
VAL
A
278
−24.407
7.674
−12.541
1.00
11.95
C


ATOM
2176
O
VAL
A
278
−24.734
7.391
−11.387
1.00
11.20
O


ATOM
2177
CB
VAL
A
278
−22.169
7.242
−13.601
1.00
9.03
C


ATOM
2178
CG1
VAL
A
278
−21.597
7.780
−12.299
1.00
12.15
C


ATOM
2179
CG2
VAL
A
278
−21.269
6.158
−14.196
1.00
13.85
C


ATOM
2180
N
THR
A
279
−24.757
8.814
−13.119
1.00
9.26
N


ATOM
2181
CA
THR
A
279
−25.478
9.838
−12.372
1.00
10.05
C


ATOM
2182
C
THR
A
279
−26.818
9.334
−11.848
1.00
9.97
C


ATOM
2183
O
THR
A
279
−27.201
9.644
−10.721
1.00
10.35
O


ATOM
2184
CB
THR
A
279
−25.640
11.119
−13.200
1.00
10.89
C


ATOM
2185
OG1
THR
A
279
−24.336
11.655
−13.456
1.00
15.41
O


ATOM
2186
CG2
THR
A
279
−26.464
12.144
−12.443
1.00
11.76
C


ATOM
2187
N
ASN
A
280
−27.534
8.562
−12.660
1.00
10.65
N


ATOM
2188
CA
ASN
A
280
−28.815
8.023
−12.224
1.00
11.97
C


ATOM
2189
C
ASN
A
280
−28.632
7.108
−11.016
1.00
9.96
C


ATOM
2190
O
ASN
A
280
−29.428
7.140
−10.067
1.00
10.76
O


ATOM
2191
CB
ASN
A
280
−29.510
7.269
−13.360
1.00
11.47
C


ATOM
2192
CG
ASN
A
280
−30.038
8.195
−14.441
1.00
21.55
C


ATOM
2193
OD1
ASN
A
280
−30.206
9.391
−14.221
1.00
21.86
O


ATOM
2194
ND2
ASN
A
280
−30.305
7.639
−15.617
1.00
25.48
N


ATOM
2195
N
SER
A
281
−27.577
6.303
−11.056
1.00
9.62
N


ATOM
2196
CA
SER
A
281
−27.240
5.423
−9.943
1.00
9.05
C


ATOM
2197
C
SER
A
281
−26.862
6.227
−8.703
1.00
10.69
C


ATOM
2198
O
SER
A
281
−27.311
5.922
−7.607
1.00
10.10
O


ATOM
2199
CB
SER
A
281
−26.089
4.494
−10.331
1.00
9.22
C


ATOM
2200
OG
SER
A
281
−25.601
3.798
−9.193
1.00
11.41
O


ATOM
2201
N
VAL
A
282
−26.029
7.250
−8.878
1.00
8.01
N


ATOM
2202
CA
VAL
A
282
−25.625
8.113
−7.769
1.00
9.36
C


ATOM
2203
C
VAL
A
282
−26.851
8.703
−7.059
1.00
8.59
C


ATOM
2204
O
VAL
A
282
−26.955
8.650
−5.827
1.00
8.36
O


ATOM
2205
CB
VAL
A
282
−24.668
9.223
−8.262
1.00
8.18
C


ATOM
2206
CG1
VAL
A
282
−24.550
10.343
−7.228
1.00
8.83
C


ATOM
2207
CG2
VAL
A
282
−23.313
8.620
−8.597
1.00
9.21
C


ATOM
2208
N
PHE
A
283
−27.787
9.254
−7.828
1.00
7.90
N


ATOM
2209
CA
PHE
A
283
−29.005
9.804
−7.247
1.00
7.68
C


ATOM
2210
C
PHE
A
283
−29.799
8.730
−6.507
1.00
10.02
C


ATOM
2211
O
PHE
A
283
−30.290
8.961
−5.403
1.00
10.89
O


ATOM
2212
CB
PHE
A
283
−29.881
10.463
−8.316
1.00
8.60
C


ATOM
2213
CG
PHE
A
283
−29.534
11.907
−8.593
1.00
8.21
C


ATOM
2214
CD1
PHE
A
283
−28.295
12.257
−9.084
1.00
9.31
C


ATOM
2215
CD2
PHE
A
283
−30.478
12.907
−8.395
1.00
9.23
C


ATOM
2216
CE1
PHE
A
283
−27.977
13.595
−9.350
1.00
10.77
C


ATOM
2217
CE2
PHE
A
283
−30.172
14.238
−8.666
1.00
10.37
C


ATOM
2218
CZ
PHE
A
283
−28.929
14.575
−9.137
1.00
10.84
C


ATOM
2219
N
HIS
A
284
−29.932
7.556
−7.121
1.00
9.09
N


ATOM
2220
CA
HIS
A
284
−30.675
6.463
−6.508
1.00
9.12
C


ATOM
2221
C
HIS
A
284
−30.033
6.025
−5.189
1.00
9.99
C


ATOM
2222
O
HIS
A
284
−30.737
5.828
−4.188
1.00
9.74
O


ATOM
2223
CB
HIS
A
284
−30.765
5.290
−7.492
1.00
11.67
C


ATOM
2224
CG
HIS
A
284
−31.431
4.069
−6.935
1.00
14.61
C


ATOM
2225
ND1
HIS
A
284
−32.760
4.046
−6.573
1.00
20.82
N


ATOM
2226
CD2
HIS
A
284
−30.958
2.820
−6.717
1.00
21.94
C


ATOM
2227
CE1
HIS
A
284
−33.074
2.837
−6.138
1.00
19.80
C


ATOM
2228
NE2
HIS
A
284
−31.998
2.075
−6.216
1.00
23.11
N


ATOM
2229
N
GLU
A
285
−28.713
5.876
−5.181
1.00
8.55
N


ATOM
2230
CA
GLU
A
285
−28.016
5.410
−3.977
1.00
8.60
C


ATOM
2231
C
GLU
A
285
−28.134
6.419
−2.836
1.00
9.27
C


ATOM
2232
O
GLU
A
285
−28.337
6.043
−1.678
1.00
10.15
O


ATOM
2233
CB
GLU
A
285
−26.548
5.102
−4.266
1.00
10.27
C


ATOM
2234
CG
GLU
A
285
−26.341
3.975
−5.284
1.00
12.66
C


ATOM
2235
CD
GLU
A
285
−26.784
2.615
−4.761
1.00
48.46
C


ATOM
2236
OE1
GLU
A
285
−26.861
2.438
−3.526
1.00
24.33
O


ATOM
2237
OE2
GLU
A
285
−27.051
1.714
−5.586
1.00
26.17
O


ATOM
2238
N
ILE
A
286
−27.999
7.703
−3.158
1.00
8.06
N


ATOM
2239
CA
ILE
A
286
−28.130
8.738
−2.137
1.00
7.60
C


ATOM
2240
C
ILE
A
286
−29.561
8.758
−1.615
1.00
8.03
C


ATOM
2241
O
ILE
A
286
−29.797
8.844
−0.404
1.00
9.29
O


ATOM
2242
CB
ILE
A
286
−27.726
10.117
−2.697
1.00
7.44
C


ATOM
2243
CG1
ILE
A
286
−26.225
10.131
−2.982
1.00
8.11
C


ATOM
2244
CG2
ILE
A
286
−28.104
11.222
−1.702
1.00
9.25
C


ATOM
2245
CD1
ILE
A
286
−25.730
11.355
−3.736
1.00
8.46
C


ATOM
2246
N
ASN
A
287
−30.525
8.668
−2.524
1.00
7.56
N


ATOM
2247
CA
ASN
A
287
−31.927
8.632
−2.145
1.00
9.10
C


ATOM
2248
C
ASN
A
287
−32.196
7.506
−1.135
1.00
9.78
C


ATOM
2249
O
ASN
A
287
−32.777
7.731
−0.068
1.00
10.92
O


ATOM
2250
CB
ASN
A
287
−32.792
8.481
−3.408
1.00
9.77
C


ATOM
2251
CG
ASN
A
287
−34.280
8.468
−3.121
1.00
16.23
C


ATOM
2252
OD1
ASN
A
287
−34.738
7.846
−2.167
1.00
16.90
O


ATOM
2253
ND2
ASN
A
287
−35.046
9.144
−3.968
1.00
17.83
N


ATOM
2254
N
MET
A
288
−31.756
6.295
−1.457
1.00
9.68
N


ATOM
2255
CA
AMET
A
288
−32.008
5.164
−0.571
0.75
10.52
C


ATOM
2256
C
MET
A
288
−31.283
5.304
0.765
1.00
10.26
C


ATOM
2257
O
MET
A
288
−31.858
5.002
1.817
1.00
11.11
O


ATOM
2258
CB
AMET
A
288
−31.618
3.853
−1.246
0.75
13.43
C


ATOM
2259
CG
AMET
A
288
−32.543
3.452
−2.384
0.75
11.44
C


ATOM
2260
SD
AMET
A
288
−32.159
1.788
−2.967
0.75
19.23
S


ATOM
2261
CE
AMET
A
288
−30.435
1.981
−3.415
0.75
25.05
C


ATOM
2262
CA
BMET
A
288
−31.970
5.141
−0.586
0.25
13.65
C


ATOM
2263
CB
BMET
A
288
−31.454
3.872
−1.264
0.25
10.56
C


ATOM
2264
CG
BMET
A
288
−32.177
3.528
−2.550
0.25
11.01
C


ATOM
2265
SD
BMET
A
288
−33.911
3.126
−2.263
0.25
15.84
S


ATOM
2266
CE
BMET
A
288
−33.757
1.686
−1.209
0.25
35.78
C


ATOM
2267
N
TRP
A
289
−30.041
5.768
0.732
1.00
8.12
N


ATOM
2268
CA
TRP
A
289
−29.239
5.874
1.947
1.00
8.46
C


ATOM
2269
C
TRP
A
289
−29.847
6.893
2.916
1.00
10.60
C


ATOM
2270
O
TRP
A
289
−30.013
6.619
4.106
1.00
10.10
O


ATOM
2271
CB
TRP
A
289
−27.809
6.262
1.588
1.00
8.04
C


ATOM
2272
CG
TRP
A
289
−26.829
6.080
2.699
1.00
8.76
C


ATOM
2273
CD1
TRP
A
289
−26.110
4.953
2.987
1.00
8.96
C


ATOM
2274
CD2
TRP
A
289
−26.452
7.057
3.681
1.00
7.64
C


ATOM
2275
NE1
TRP
A
289
−25.307
5.171
4.076
1.00
8.96
N


ATOM
2276
CE2
TRP
A
289
−25.501
6.452
4.525
1.00
7.40
C


ATOM
2277
CE3
TRP
A
289
−26.815
8.386
3.916
1.00
8.72
C


ATOM
2278
CZ2
TRP
A
289
−24.912
7.130
5.594
1.00
8.56
C


ATOM
2279
CZ3
TRP
A
289
−26.232
9.055
4.981
1.00
9.78
C


ATOM
2280
CH2
TRP
A
289
−25.292
8.429
5.801
1.00
7.77
C


ATOM
2281
N
VAL
A
290
−30.202
8.061
2.392
1.00
7.61
N


ATOM
2282
CA
VAL
A
290
−30.822
9.098
3.210
1.00
8.87
C


ATOM
2283
C
VAL
A
290
−32.228
8.697
3.658
1.00
11.01
C


ATOM
2284
O
VAL
A
290
−32.591
8.898
4.821
1.00
12.42
O


ATOM
2285
CB
VAL
A
290
−30.834
10.451
2.465
1.00
10.20
C


ATOM
2286
CG1
VAL
A
290
−31.622
11.483
3.254
1.00
13.69
C


ATOM
2287
CG2
VAL
A
290
−29.410
10.923
2.232
1.00
10.54
C


ATOM
2288
N
SER
A
291
−33.018
8.122
2.757
1.00
11.05
N


ATOM
2289
CA
SER
A
291
−34.362
7.653
3.105
1.00
12.68
C


ATOM
2290
C
SER
A
291
−34.337
6.683
4.286
1.00
15.97
C


ATOM
2291
O
SER
A
291
−35.128
6.805
5.226
1.00
16.56
O


ATOM
2292
CB
SER
A
291
−35.013
6.968
1.903
1.00
14.10
C


ATOM
2293
OG
SER
A
291
−35.461
7.922
0.957
1.00
20.61
O


ATOM
2294
N
GLN
A
292
−33.428
5.717
4.236
1.00
13.80
N


ATOM
2295
CA
GLN
A
292
−33.346
4.702
5.281
1.00
13.65
C


ATOM
2296
C
GLN
A
292
−32.926
5.283
6.628
1.00
15.92
C


ATOM
2297
O
GLN
A
292
−33.192
4.697
7.676
1.00
19.93
O


ATOM
2298
CB
GLN
A
292
−32.414
3.577
4.841
1.00
15.28
C


ATOM
2299
CG
GLN
A
292
−33.016
2.750
3.714
1.00
31.36
C


ATOM
2300
CD
GLN
A
292
−32.018
1.827
3.059
1.00
59.35
C


ATOM
2301
OE1
GLN
A
292
−31.042
1.409
3.679
1.00
46.54
O


ATOM
2302
NE2
GLN
A
292
−32.259
1.500
1.795
1.00
52.18
N


ATOM
2303
N
ARG
A
293
−32.290
6.449
6.604
1.00
11.83
N


ATOM
2304
CA
ARG
A
293
−31.758
7.049
7.818
1.00
11.03
C


ATOM
2305
C
ARG
A
293
−32.577
8.254
8.293
1.00
10.89
C


ATOM
2306
O
ARG
A
293
−32.172
8.968
9.215
1.00
14.96
O


ATOM
2307
CB
ARG
A
293
−30.273
7.380
7.632
1.00
9.52
C


ATOM
2308
CG
ARG
A
293
−29.406
6.126
7.665
1.00
10.01
C


ATOM
2309
CD
ARG
A
293
−28.042
6.325
7.036
1.00
10.98
C


ATOM
2310
NE
ARG
A
293
−27.297
5.065
7.004
1.00
9.60
N


ATOM
2311
CZ
ARG
A
293
−27.508
4.099
6.111
1.00
9.62
C


ATOM
2312
NH1
ARG
A
293
−28.431
4.249
5.166
1.00
9.87
N


ATOM
2313
NH2
ARG
A
293
−26.789
2.987
6.171
1.00
11.78
N


ATOM
2314
N
THR
A
294
−33.731
8.453
7.667
1.00
14.09
N


ATOM
2315
CA
THR
A
294
−34.681
9.485
8.068
1.00
15.85
C


ATOM
2316
C
THR
A
294
−36.072
8.869
8.199
1.00
22.42
C


ATOM
2317
O
THR
A
294
−37.067
9.584
8.311
1.00
36.87
O


ATOM
2318
CB
THR
A
294
−34.731
10.651
7.053
1.00
16.73
C


ATOM
2319
OG1
THR
A
294
−35.000
10.137
5.743
1.00
18.85
O


ATOM
2320
CG2
THR
A
294
−33.417
11.393
7.022
1.00
15.10
C


ATOM
2321
O01
INH
I
1
−12.895
20.823
−4.999
1.00
7.09
O


ATOM
2322
C02
INH
I
1
−12.391
20.703
−6.129
1.00
8.09
C


ATOM
2323
C03
INH
I
1
−10.917
20.735
−6.329
1.00
7.66
C


ATOM
2324
C04
INH
I
1
−10.371
21.511
−7.372
1.00
8.80
C


ATOM
2325
C05
INH
I
1
−9.002
21.548
−7.621
1.00
11.29
C


ATOM
2326
C06
INH
I
1
−8.161
20.785
−6.808
1.00
9.74
C


ATOM
2327
N07
INH
I
1
−6.763
20.578
−6.783
1.00
12.42
N


ATOM
2328
C08
INH
I
1
−6.533
19.727
−5.790
1.00
14.65
C


ATOM
2329
O09
INH
I
1
−7.712
19.334
−5.112
1.00
13.07
O


ATOM
2330
C10
INH
I
1
−8.755
19.997
−5.760
1.00
9.91
C


ATOM
2331
C11
INH
I
1
−10.110
19.953
−5.499
1.00
7.61
C


ATOM
2332
C12
INH
I
1
−5.241
19.125
−5.317
1.00
16.69
C


ATOM
2333
C13
INH
I
1
−4.117
19.335
−6.328
1.00
15.09
C


ATOM
2334
C14
INH
I
1
−2.767
18.863
−5.823
1.00
19.81
C


ATOM
2335
C15
INH
I
1
−2.421
19.447
−4.476
1.00
19.20
C


ATOM
2336
C16
INH
I
1
−3.512
19.142
−3.467
1.00
19.60
C


ATOM
2337
C17
INH
I
1
−4.893
19.592
−3.914
1.00
17.98
C


ATOM
2338
N18
INH
I
1
−13.235
20.690
−7.197
1.00
6.87
N


ATOM
2339
C19
INH
I
1
−14.672
20.540
−7.283
1.00
6.93
C


ATOM
2340
C20
INH
I
1
−14.471
19.893
−8.684
1.00
6.67
C


ATOM
2341
C21
INH
I
1
−12.952
20.266
−8.587
1.00
8.77
C


ATOM
2342
N22
INH
I
1
−15.178
20.424
−9.813
1.00
6.45
N


ATOM
2343
C23
INH
I
1
−16.588
20.190
−9.789
1.00
5.98
C


ATOM
2344
C24
INH
I
1
−17.328
20.733
−11.042
1.00
6.80
C


ATOM
2345
N25
INH
I
1
−16.634
20.331
−12.248
1.00
7.33
N


ATOM
2346
C26
INH
I
1
−15.203
20.541
−12.284
1.00
10.22
C


ATOM
2347
C27
INH
I
1
−14.523
19.973
−11.038
1.00
7.33
C


ATOM
2348
C28
INH
I
1
−17.296
19.725
−13.318
1.00
6.80
C


ATOM
2349
N29
INH
I
1
−16.488
19.205
−14.329
1.00
7.30
N


ATOM
2350
C30
INH
I
1
−17.138
18.618
−15.336
1.00
7.90
C


ATOM
2351
C31
INH
I
1
−18.527
18.508
−15.408
1.00
7.42
C


ATOM
2352
C32
INH
I
1
−19.257
19.048
−14.354
1.00
7.83
C


ATOM
2353
N33
INH
I
1
−18.670
19.648
−13.307
1.00
6.45
N


HETATM
2354
O
HOH
W
1
−17.954
27.691
4.082
1.00
7.52
O


HETATM
2355
O
HOH
W
2
−16.687
30.118
3.647
1.00
7.58
O


HETATM
2356
O
HOH
W
3
−16.903
29.964
−7.861
1.00
8.56
O


HETATM
2357
O
HOH
W
4
−24.521
30.541
−6.460
1.00
8.76
O


HETATM
2358
O
HOH
W
5
−18.264
32.412
10.389
1.00
8.02
O


HETATM
2359
O
HOH
W
6
−23.247
3.675
5.398
1.00
9.13
O


HETATM
2360
O
HOH
W
7
−15.105
23.282
−9.818
1.00
10.08
O


HETATM
2361
O
HOH
W
8
−20.744
0.215
−8.443
1.00
9.73
O


HETATM
2362
O
HOH
W
9
−11.897
30.578
−1.691
1.00
10.64
O


HETATM
2363
O
HOH
W
10
−22.252
20.341
−15.373
1.00
11.45
O


HETATM
2364
O
HOH
W
11
−27.187
18.469
−8.872
1.00
12.11
O


HETATM
2365
O
HOH
W
12
−11.733
34.673
−0.588
1.00
12.31
O


HETATM
2366
O
HOH
W
13
−11.423
12.123
−13.304
1.00
11.25
O


HETATM
2367
O
HOH
W
14
−18.816
28.913
13.424
1.00
11.56
O


HETATM
2368
O
HOH
W
15
−9.196
10.830
−2.094
1.00
12.11
O


HETATM
2369
O
HOH
W
16
−9.803
26.896
−12.434
1.00
12.21
O


HETATM
2370
O
HOH
W
17
−21.490
5.488
12.067
1.00
12.81
O


HETATM
2371
O
HOH
W
18
−32.761
29.987
−2.012
1.00
14.02
O


HETATM
2372
O
HOH
W
19
−18.414
31.657
13.060
1.00
12.16
O


HETATM
2373
O
HOH
W
20
−5.462
30.956
−0.225
1.00
13.69
O


HETATM
2374
O
HOH
W
21
−11.248
35.923
1.786
1.00
14.53
O


HETATM
2375
O
HOH
W
22
−30.857
31.479
−3.105
1.00
12.95
O


HETATM
2376
O
HOH
W
23
−17.537
30.497
−17.209
1.00
13.52
O


HETATM
2377
O
HOH
W
24
−27.943
15.491
13.891
1.00
15.19
O


HETATM
2378
O
HOH
W
25
−6.754
29.942
−6.600
1.00
14.74
O


HETATM
2379
O
HOH
W
26
−21.995
24.699
−0.661
1.00
15.81
O


HETATM
2380
O
HOH
W
27
−24.620
37.557
−8.143
1.00
14.70
O


HETATM
2381
O
HOH
W
28
−11.836
10.105
−2.131
1.00
13.84
O


HETATM
2382
O
HOH
W
29
−6.977
17.152
−20.873
1.00
14.48
O


HETATM
2383
O
HOH
W
30
−27.778
25.779
17.062
1.00
14.55
O


HETATM
2384
O
HOH
W
31
−28.099
20.389
−10.972
1.00
15.25
O


HETATM
2385
O
HOH
W
32
−26.572
16.678
−10.935
1.00
15.41
O


HETATM
2386
O
HOH
W
33
−21.236
20.183
−9.116
1.00
14.32
O


HETATM
2387
O
HOH
W
34
−21.239
28.606
14.850
1.00
15.96
O


HETATM
2388
O
HOH
W
35
−24.773
29.548
−12.422
1.00
15.43
O


HETATM
2389
O
HOH
W
36
−20.822
20.806
−11.819
1.00
14.61
O


HETATM
2390
O
HOH
W
37
−9.275
15.936
−22.055
1.00
14.71
O


HETATM
2391
O
HOH
W
38
−20.594
14.589
13.855
1.00
17.78
O


HETATM
2392
O
HOH
W
39
5.208
15.094
−10.204
1.00
16.41
O


HETATM
2393
O
HOH
W
40
−20.072
5.193
14.956
1.00
15.13
O


HETATM
2394
O
HOH
W
41
−13.159
26.020
−13.404
1.00
16.62
O


HETATM
2395
O
HOH
W
42
−26.282
23.518
16.511
1.00
20.84
O


HETATM
2396
O
HOH
W
43
−10.302
34.220
3.868
1.00
18.17
O


HETATM
2397
O
HOH
W
44
−1.928
29.772
2.061
1.00
15.99
O


HETATM
2398
O
HOH
W
45
−2.744
28.535
7.126
1.00
19.79
O


HETATM
2399
O
HOH
W
46
−5.515
17.879
−17.351
1.00
19.09
O


HETATM
2400
O
HOH
W
47
−12.817
5.509
−17.997
1.00
18.20
O


HETATM
2401
O
HOH
W
48
−14.357
31.242
−7.410
1.00
17.27
O


HETATM
2402
O
HOH
W
49
−14.923
14.930
−21.033
1.00
17.02
O


HETATM
2403
O
HOH
W
50
−9.458
23.156
9.139
1.00
21.87
O


HETATM
2404
O
HOH
W
51
0.257
31.525
−7.428
1.00
18.65
O


HETATM
2405
O
HOH
W
52
−25.578
20.460
−14.720
1.00
18.62
O


HETATM
2406
O
HOH
W
53
−20.547
12.617
−23.501
1.00
20.46
O


HETATM
2407
O
HOH
W
54
−23.950
26.787
−18.563
1.00
17.02
O


HETATM
2408
O
HOH
W
55
−14.696
9.975
−8.189
1.00
19.62
O


HETATM
2409
O
HOH
W
56
−25.051
13.877
−15.630
1.00
17.92
O


HETATM
2410
O
HOH
W
57
−8.564
7.506
−8.708
1.00
18.44
O


HETATM
2411
O
HOH
W
58
−26.198
31.102
−16.423
1.00
16.37
O


HETATM
2412
O
HOH
W
59
−37.176
12.986
−4.281
1.00
21.18
O


HETATM
2413
O
HOH
W
60
−0.408
19.850
−9.658
1.00
20.96
O


HETATM
2414
O
HOH
W
61
−27.390
2.115
−8.137
1.00
20.41
O


HETATM
2415
O
HOH
W
62
−18.345
0.114
10.794
1.00
19.73
O


HETATM
2416
O
HOH
W
63
−7.560
17.472
11.844
1.00
20.82
O


HETATM
2417
O
HOH
W
64
−10.060
27.021
−17.033
1.00
19.15
O


HETATM
2418
O
HOH
W
65
−11.889
25.550
−15.567
1.00
17.58
O


HETATM
2419
O
HOH
W
66
−0.270
32.535
−4.292
1.00
23.13
O


HETATM
2420
O
HOH
W
67
−2.409
13.621
−0.840
1.00
20.38
O


HETATM
2421
O
HOH
W
68
−26.262
31.478
−13.675
1.00
20.59
O


HETATM
2422
O
HOH
W
69
−28.229
3.738
−13.695
1.00
21.62
O


HETATM
2423
O
HOH
W
70
−31.712
27.290
−6.129
1.00
18.31
O


HETATM
2424
O
HOH
W
71
−6.300
21.250
−19.853
1.00
16.05
O


HETATM
2425
O
HOH
W
72
−28.617
2.134
3.116
1.00
24.09
O


HETATM
2426
O
HOH
W
73
−6.265
12.778
−21.032
1.00
20.88
O


HETATM
2427
O
HOH
W
74
−5.345
21.003
−9.170
1.00
20.95
O


HETATM
2428
O
HOH
W
75
−13.672
40.032
1.481
1.00
20.82
O


HETATM
2429
O
HOH
W
76
3.970
33.284
−8.733
1.00
21.52
O


HETATM
2430
O
HOH
W
77
−27.601
3.453
−1.085
1.00
19.94
O


HETATM
2431
O
HOH
W
78
−16.463
32.339
14.904
1.00
20.41
O


HETATM
2432
O
HOH
W
79
−30.731
36.819
4.054
1.00
22.74
O


HETATM
2433
O
HOH
W
80
−10.846
1.831
−7.270
1.00
21.37
O


HETATM
2434
O
HOH
W
81
−27.313
22.838
−11.310
1.00
24.03
O


HETATM
2435
O
HOH
W
82
−15.217
31.789
−19.315
1.00
23.88
O


HETATM
2436
O
HOH
W
83
−33.912
16.169
−9.052
1.00
22.87
O


HETATM
2437
O
HOH
W
84
−20.209
−1.679
12.463
1.00
21.35
O


HETATM
2438
O
HOH
W
85
−5.913
10.247
−9.105
1.00
22.32
O


HETATM
2439
O
HOH
W
86
−0.959
17.408
2.354
1.00
20.98
O


HETATM
2440
O
HOH
W
87
−10.503
38.192
0.316
1.00
24.62
O


HETATM
2441
O
HOH
W
88
−32.112
7.871
−10.243
1.00
21.94
O


HETATM
2442
O
HOH
W
89
−13.180
25.290
−17.756
1.00
19.54
O


HETATM
2443
O
HOH
W
90
−7.804
19.778
−13.041
1.00
24.06
O


HETATM
2444
O
HOH
W
91
−33.427
27.311
−4.139
1.00
23.94
O


HETATM
2445
O
HOH
W
92
−12.229
34.941
5.552
1.00
22.55
O


HETATM
2446
O
HOH
W
93
−40.268
22.945
5.322
1.00
24.66
O


HETATM
2447
O
HOH
W
94
−11.361
−0.276
9.385
1.00
24.33
O


HETATM
2448
O
HOH
W
95
−5.594
12.673
4.154
1.00
21.98
O


HETATM
2449
O
HOH
W
96
−30.277
8.393
11.189
1.00
23.95
O


HETATM
2450
O
HOH
W
97
−3.885
31.646
2.132
1.00
23.08
O


HETATM
2451
O
HOH
W
98
−15.349
37.712
−10.155
1.00
22.78
O


HETATM
2452
O
HOH
W
99
−27.793
8.032
10.121
1.00
21.52
O


HETATM
2453
O
HOH
W
100
1.726
25.595
−13.323
1.00
24.61
O


HETATM
2454
O
HOH
W
101
−4.022
23.204
−10.234
1.00
23.85
O


HETATM
2455
O
HOH
W
102
−0.283
27.133
5.706
1.00
27.07
O


HETATM
2456
O
HOH
W
103
−17.612
12.195
13.686
1.00
22.47
O


HETATM
2457
O
HOH
W
104
−20.617
38.511
11.379
1.00
22.85
O


HETATM
2458
O
HOH
W
105
−12.949
21.628
−20.519
1.00
22.65
O


HETATM
2459
O
HOH
W
106
−8.950
13.120
−21.827
1.00
24.72
O


HETATM
2460
O
HOH
W
107
−24.860
6.230
−19.460
1.00
22.27
O


HETATM
2461
O
HOH
W
108
−19.286
25.663
−23.124
1.00
24.09
O


HETATM
2462
O
HOH
W
109
−4.483
32.563
−11.039
1.00
27.41
O


HETATM
2463
O
HOH
W
110
−29.993
33.249
4.804
1.00
24.16
O


HETATM
2464
O
HOH
W
111
12.207
32.873
0.335
1.00
25.14
O


HETATM
2465
O
HOH
W
112
−34.084
27.205
19.146
1.00
21.94
O


HETATM
2466
O
HOH
W
113
−26.787
5.452
10.766
1.00
22.13
O


HETATM
2467
O
HOH
W
114
1.550
16.445
−12.673
1.00
29.27
O


HETATM
2468
O
HOH
W
115
0.773
22.236
−9.903
1.00
21.44
O


HETATM
2469
O
HOH
W
116
−15.044
17.209
−22.620
1.00
21.87
O


HETATM
2470
O
HOH
W
117
−25.901
32.220
9.915
1.00
21.85
O


HETATM
2471
O
HOH
W
118
−15.258
40.579
−2.244
1.00
27.72
O


HETATM
2472
O
HOH
W
119
−7.794
30.680
−9.041
1.00
23.66
O


HETATM
2473
O
HOH
W
120
−18.635
35.461
−11.584
1.00
24.31
O


HETATM
2474
O
HOH
W
121
−26.081
16.654
−25.505
1.00
22.43
O


HETATM
2475
O
HOH
W
122
−9.221
9.291
−15.768
1.00
27.18
O


HETATM
2476
O
HOH
W
123
−28.859
32.651
−7.827
1.00
31.44
O


HETATM
2477
O
HOH
W
124
−1.147
11.892
−2.485
1.00
25.34
O


HETATM
2478
O
HOH
W
125
−16.717
23.990
−23.206
1.00
22.72
O


HETATM
2479
O
HOH
W
126
−16.904
27.933
15.146
1.00
28.48
O


HETATM
2480
O
HOH
W
127
−24.881
−0.157
−9.675
1.00
30.44
O


HETATM
2481
O
HOH
W
128
−12.528
32.808
−13.738
1.00
23.97
O


HETATM
2482
O
HOH
W
129
−31.526
33.432
−4.910
1.00
29.01
O


HETATM
2483
O
HOH
W
130
−10.048
37.820
−7.038
1.00
29.37
O


HETATM
2484
O
HOH
W
131
−11.867
19.403
12.228
1.00
29.42
O


HETATM
2485
O
HOH
W
132
−37.133
10.668
3.881
1.00
30.27
O


HETATM
2486
O
HOH
W
133
−19.747
3.815
−16.456
1.00
27.29
O


HETATM
2487
O
HOH
W
134
−7.880
35.404
−6.844
1.00
24.86
O


HETATM
2488
O
HOH
W
135
−8.998
3.716
−9.036
1.00
28.14
O


HETATM
2489
O
HOH
W
136
−2.967
12.137
4.048
1.00
26.40
O


HETATM
2490
O
HOH
W
137
−17.049
39.326
−11.678
1.00
25.80
O


HETATM
2491
O
HOH
W
138
−10.257
−2.109
−12.040
1.00
29.56
O


HETATM
2492
O
HOH
W
139
−26.119
34.050
−13.125
1.00
25.74
O


HETATM
2493
O
HOH
W
140
−16.994
38.359
14.121
1.00
32.67
O


HETATM
2494
O
HOH
W
141
−10.346
37.403
−10.034
1.00
28.07
O


HETATM
2495
O
HOH
W
142
6.226
22.804
−1.176
1.00
26.21
O


HETATM
2496
O
HOH
W
143
−7.855
32.548
9.121
1.00
31.08
O


HETATM
2497
O
HOH
W
144
−38.520
18.008
−5.332
1.00
28.74
O


HETATM
2498
O
HOH
W
145
0.863
30.021
−14.155
1.00
25.22
O


HETATM
2499
O
HOH
W
146
−14.539
12.832
−22.506
1.00
34.74
O


HETATM
2500
O
HOH
W
147
−9.685
35.266
7.783
1.00
39.75
O


HETATM
2501
O
HOH
W
148
−4.940
15.663
−7.095
1.00
28.86
O


HETATM
2502
O
HOH
W
149
−19.875
12.007
14.866
1.00
29.93
O


HETATM
2503
O
HOH
W
150
−25.128
−0.534
−5.095
1.00
28.37
O


HETATM
2504
O
HOH
W
151
−1.385
12.790
1.731
1.00
27.27
O


HETATM
2505
O
HOH
W
152
−28.589
35.688
−8.140
1.00
29.37
O


HETATM
2506
O
HOH
W
153
−39.946
16.242
11.666
1.00
27.63
O


HETATM
2507
O
HOH
W
154
−12.005
32.563
12.899
1.00
28.89
O


HETATM
2508
O
HOH
W
155
−32.382
11.270
10.718
1.00
27.03
O


HETATM
2509
O
HOH
W
156
−6.547
26.498
11.420
1.00
26.38
O


HETATM
2510
O
HOH
W
157
−15.098
4.994
−20.108
1.00
30.09
O


HETATM
2511
O
HOH
W
158
−40.304
25.037
6.932
1.00
29.28
O


HETATM
2512
O
HOH
W
159
−19.654
42.452
−11.933
1.00
28.67
O


HETATM
2513
O
HOH
W
160
7.165
33.655
−14.239
1.00
29.40
O


HETATM
2514
O
HOH
W
161
−29.918
4.810
−15.685
1.00
30.16
O


HETATM
2515
O
HOH
W
162
10.935
23.184
−6.110
1.00
32.43
O


HETATM
2516
O
HOH
W
163
−26.145
2.186
1.054
1.00
36.44
O


HETATM
2517
O
HOH
W
164
−34.339
6.200
−6.038
1.00
31.60
O


HETATM
2518
O
HOH
W
165
−7.577
23.852
11.873
1.00
37.15
O


HETATM
2519
O
HOH
W
166
−28.374
17.766
15.757
1.00
33.08
O


HETATM
2520
O
HOH
W
167
−8.878
30.353
10.450
1.00
33.65
O


HETATM
2521
O
HOH
W
168
−39.240
24.455
13.603
1.00
32.18
O


HETATM
2522
O
HOH
W
169
−5.259
32.467
−14.521
1.00
30.28
O


HETATM
2523
O
HOH
W
170
−19.002
44.014
−6.429
1.00
36.88
O


HETATM
2524
O
HOH
W
171
−41.472
23.045
−1.746
1.00
34.68
O


HETATM
2525
O
HOH
W
172
−41.240
17.380
−0.287
1.00
30.69
O


HETATM
2526
O
HOH
W
173
−18.871
43.325
−2.077
1.00
32.89
O


HETATM
2527
O
HOH
W
174
−15.823
41.421
2.253
1.00
29.64
O


HETATM
2528
O
HOH
W
175
−7.050
21.485
−11.124
1.00
32.96
O


HETATM
2529
O
HOH
W
176
−37.741
26.386
−1.056
1.00
29.25
O


HETATM
2530
O
HOH
W
177
−37.633
14.168
12.267
1.00
41.58
O


HETATM
2531
O
HOH
W
178
−36.152
30.566
14.397
1.00
32.08
O


HETATM
2532
O
HOH
W
179
−16.547
10.926
−22.860
1.00
29.03
O


HETATM
2533
O
HOH
W
180
−41.727
23.706
9.092
1.00
33.88
O


HETATM
2534
O
HOH
W
181
6.999
23.263
−12.013
1.00
36.81
O


HETATM
2535
O
HOH
W
182
−17.044
8.041
−22.415
1.00
38.12
O


HETATM
2536
O
HOH
W
183
−35.338
12.042
10.329
1.00
31.94
O


HETATM
2537
O
HOH
W
184
2.050
23.137
−12.328
1.00
30.75
O


HETATM
2538
O
HOH
W
185
−19.726
45.933
−2.467
1.00
38.74
O


HETATM
2539
O
HOH
W
186
−0.301
9.788
−9.818
1.00
33.75
O


HETATM
2540
O
HOH
W
187
−5.231
11.780
14.591
1.00
34.83
O


HETATM
2541
O
HOH
W
188
−14.916
41.932
5.514
1.00
41.07
O


HETATM
2542
O
HOH
W
189
−42.696
19.572
−0.424
1.00
32.59
O


HETATM
2543
O
HOH
W
190
−0.895
14.955
4.198
1.00
36.30
O


HETATM
2544
O
HOH
W
191
−18.995
3.659
−13.597
1.00
34.12
O


HETATM
2545
O
HOH
W
192
−37.998
28.389
15.014
1.00
31.37
O


HETATM
2546
O
HOH
W
193
−30.890
10.641
13.026
1.00
32.22
O


HETATM
2547
O
HOH
W
194
−11.626
11.766
−21.687
1.00
36.98
O


HETATM
2548
O
HOH
W
195
−14.658
39.550
−8.121
1.00
38.81
O


HETATM
2549
O
HOH
W
196
−29.019
13.051
14.830
1.00
36.98
O


HETATM
2550
O
HOH
W
197
−4.504
18.598
−10.348
1.00
38.67
O


HETATM
2551
O
HOH
W
198
−0.549
14.853
−12.387
1.00
44.66
O


HETATM
2552
O
HOH
W
199
−4.956
22.211
−15.525
1.00
36.99
O


HETATM
2553
O
HOH
W
200
−4.889
15.947
−10.022
1.00
35.93
O


HETATM
2554
O
HOH
W
201
−23.924
21.128
−22.907
1.00
33.50
O


HETATM
2555
O
HOH
W
202
−15.047
33.244
−10.368
1.00
35.92
O


HETATM
2556
O
HOH
W
203
−38.527
11.133
10.261
1.00
37.83
O


HETATM
2557
O
HOH
W
204
−9.009
31.981
−16.860
1.00
31.39
O


HETATM
2558
O
HOH
W
205
−38.191
21.671
15.013
1.00
36.15
O


HETATM
2559
O
HOH
W
206
−18.868
10.333
−23.932
1.00
46.13
O


HETATM
2560
O
HOH
W
207
−35.132
25.919
−6.614
1.00
52.07
O


HETATM
2561
O
HOH
W
208
−27.841
21.763
−15.855
1.00
41.85
O


HETATM
2562
O
HOH
W
209
−34.644
43.404
−1.444
1.00
38.81
O


HETATM
2563
O
HOH
W
210
−30.201
3.894
−11.360
1.00
37.63
O


HETATM
2564
O
HOH
W
211
−22.748
49.399
−4.251
1.00
36.11
O


HETATM
2565
O
HOH
W
212
−1.483
14.264
8.431
1.00
41.49
O


HETATM
2566
O
HOH
W
213
−23.051
11.708
−21.920
1.00
36.32
O


HETATM
2567
O
HOH
W
214
1.620
18.299
2.516
1.00
40.18
O


HETATM
2568
O
HOH
W
215
−24.103
43.355
−12.751
1.00
37.49
O


HETATM
2569
O
HOH
W
216
−26.738
0.134
−2.192
1.00
40.25
O


HETATM
2570
O
HOH
W
217
11.242
21.332
−4.106
1.00
38.90
O


HETATM
2571
O
HOH
W
218
−14.341
33.661
14.752
1.00
43.39
O


HETATM
2572
O
HOH
W
219
−42.307
15.549
8.867
1.00
48.18
O


HETATM
2573
O
HOH
W
220
−36.432
28.737
−0.897
1.00
39.64
O


HETATM
2574
O
HOH
W
221
−25.417
28.494
−16.695
1.00
29.40
O


HETATM
2575
O
HOH
W
222
−19.511
43.409
6.951
1.00
28.17
O


HETATM
2576
O
HOH
W
223
−10.123
16.512
−24.639
1.00
31.70
O


HETATM
2577
O
HOH
W
224
−25.245
45.166
−11.086
1.00
29.38
O


HETATM
2578
O
HOH
W
225
−23.616
45.957
4.604
1.00
46.22
O


HETATM
2579
O
HOH
W
226
−4.587
19.737
−15.634
1.00
37.60
O


HETATM
2580
O
HOH
W
227
−24.137
7.274
−21.752
1.00
39.86
O


HETATM
2581
O
HOH
W
228
−12.824
13.338
18.692
1.00
35.98
O


HETATM
2582
O
HOH
W
229
−0.643
15.774
−0.049
1.00
38.98
O


HETATM
2583
O
HOH
W
230
−19.204
39.608
13.339
1.00
44.88
O


HETATM
2584
O
HOH
W
231
−40.196
18.266
13.870
1.00
39.93
O


HETATM
2585
O
HOH
W
232
−18.669
36.100
15.012
1.00
40.93
O


HETATM
2586
O
HOH
W
233
−29.410
26.628
−9.566
1.00
40.47
O


HETATM
2587
O
HOH
W
234
−30.809
34.598
−7.203
1.00
42.41
O


HETATM
2588
O
HOH
W
235
−22.479
15.485
19.837
1.00
46.47
O


HETATM
2589
O
HOH
W
236
−22.471
4.871
16.342
1.00
44.82
O


HETATM
2590
O
HOH
W
237
−34.347
29.636
−4.151
1.00
41.68
O


HETATM
2591
O
HOH
W
238
−36.466
16.190
−8.418
1.00
38.78
O


HETATM
2592
O
HOH
W
239
−17.832
41.337
−3.943
1.00
40.59
O


HETATM
2593
O
HOH
W
240
−5.599
3.373
1.035
1.00
37.39
O


HETATM
2594
O
HOH
W
241
−31.315
43.021
6.641
1.00
44.12
O


HETATM
2595
O
HOH
W
242
−0.730
9.262
−1.771
1.00
40.88
O


HETATM
2596
O
HOH
W
243
4.759
11.224
−6.950
1.00
47.42
O


HETATM
2597
O
HOH
W
244
−10.999
3.648
−16.952
1.00
46.46
O


HETATM
2598
O
HOH
W
245
−5.652
21.836
12.464
1.00
42.81
O


HETATM
2599
O
HOH
W
246
−14.550
10.917
18.018
1.00
41.09
O


HETATM
2600
O
HOH
W
247
−4.947
4.565
−7.195
1.00
46.09
O


HETATM
2601
O
HOH
W
248
−24.134
51.667
−5.997
1.00
46.36
O


HETATM
2602
O
HOH
W
249
−37.327
13.552
−6.997
1.00
53.05
O


HETATM
2603
O
HOH
W
250
0.506
22.694
3.267
1.00
47.90
O


HETATM
2604
O
HOH
W
251
−13.591
16.310
−24.807
1.00
50.85
O


HETATM
2605
O
HOH
W
252
−9.018
7.158
−17.757
1.00
46.53
O


HETATM
2606
O
HOH
W
253
−26.754
28.420
−10.600
1.00
29.06
O


HETATM
2607
O
HOH
W
254
−18.410
44.305
9.205
1.00
29.63
O


HETATM
2608
O
HOH
W
255
−32.584
10.434
−11.160
1.00
41.22
O


HETATM
2609
O
HOH
W
256
−16.091
1.690
16.162
1.00
36.96
O


HETATM
2610
O
HOH
W
257
−4.505
9.349
11.016
1.00
39.86
O


HETATM
2611
O
HOH
W
258
−40.947
18.159
−3.757
1.00
43.42
O


HETATM
2612
O
HOH
W
259
0.000
24.141
−15.150
1.00
38.32
O


HETATM
2613
O
HOH
W
260
−44.582
24.137
6.565
1.00
51.92
O


HETATM
2614
O
HOH
W
261
−35.232
13.693
−8.515
1.00
55.39
O


HETATM
2615
O
HOH
W
262
−29.872
45.235
6.694
1.00
17.67
O


HETATM
2616
O
HOH
W
263
−27.113
45.109
−0.458
1.00
22.65
O


HETATM
2617
O
HOH
W
264
−25.323
47.035
2.382
1.00
28.79
O


HETATM
2618
O
HOH
W
265
0.000
26.981
−15.150
1.00
32.82
O


HETATM
2619
O
HOH
W
266
−18.156
20.703
−27.017
1.00
35.61
O


HETATM
2620
O
HOH
W
267
−37.722
10.433
−4.365
1.00
45.06
O


HETATM
2621
O
HOH
W
268
−16.805
35.390
−9.635
1.00
40.40
O


HETATM
2622
O
HOH
W
269
−16.918
18.529
−26.281
1.00
39.96
O


HETATM
2623
O
HOH
W
270
−13.491
38.406
−6.271
1.00
39.29
O


HETATM
2624
O
HOH
W
271
−8.148
20.081
11.966
1.00
44.03
O


HETATM
2625
O
HOH
W
272
−6.821
1.644
3.126
1.00
40.41
O


HETATM
2626
O
HOH
W
273
−18.313
5.740
−22.619
1.00
47.63
O


HETATM
2627
O
HOH
W
274
−13.103
2.644
−18.422
1.00
45.18
O


HETATM
2628
O
HOH
W
275
−0.862
26.991
8.197
1.00
49.93
O


HETATM
2629
O
HOH
W
276
−12.480
35.715
13.919
1.00
48.53
O


HETATM
2630
O
HOH
W
277
−6.665
2.991
−5.698
1.00
47.86
O


HETATM
2631
O
HOH
W
278
−6.235
34.850
−10.896
1.00
49.10
O


HETATM
2632
O
HOH
W
279
−15.224
26.534
14.107
1.00
34.50
O


HETATM
2633
O
HOH
W
280
−26.785
17.717
−14.048
1.00
32.29
O


HETATM
2634
O
HOH
W
281
−10.359
3.127
−14.054
1.00
35.76
O


HETATM
2635
O
HOH
W
282
−9.964
34.823
−10.668
1.00
36.66
O


HETATM
2636
O
HOH
W
283
−10.871
37.987
5.889
1.00
39.81
O


HETATM
2637
O
HOH
W
284
−26.828
12.543
−16.696
1.00
39.23
O


HETATM
2638
O
HOH
W
285
−28.134
16.073
−12.715
1.00
39.10
O


HETATM
2639
O
HOH
W
286
−8.308
33.139
−8.609
1.00
41.47
O


HETATM
2640
O
HOH
W
287
4.685
22.519
−12.295
1.00
43.15
O


HETATM
2641
O
HOH
W
288
−6.455
17.658
−13.209
1.00
43.22
O


HETATM
2642
O
HOH
W
289
−29.264
4.527
11.185
1.00
38.52
O


HETATM
2643
O
HOH
W
290
1.948
31.540
−15.946
1.00
42.96
O


HETATM
2644
O
HOH
W
291
−28.001
47.360
−9.030
1.00
42.25
O


HETATM
2645
O
HOH
W
292
−21.578
9.488
15.696
1.00
48.22
O


HETATM
2646
O
HOH
W
293
−36.209
19.343
−11.413
1.00
43.75
O


HETATM
2647
O
HOH
W
294
0.319
29.212
4.405
1.00
42.45
O


HETATM
2648
O
HOH
W
295
−43.593
18.340
3.119
1.00
44.30
O


HETATM
2649
O
HOH
W
296
−8.690
9.796
−18.360
1.00
48.63
O


HETATM
2650
O
HOH
W
297
−13.183
20.728
−23.025
1.00
42.14
O


HETATM
2651
O
HOH
W
298
−28.554
16.113
−23.692
1.00
44.64
O


HETATM
2652
O
HOH
W
299
−11.671
39.418
10.822
1.00
44.26
O


HETATM
2653
O
HOH
W
300
−7.956
4.305
−13.456
1.00
46.03
O


HETATM
2654
O
HOH
W
301
−39.821
23.326
16.353
1.00
43.78
O


HETATM
2655
O
HOH
W
302
−13.526
31.064
14.928
1.00
53.12
O


HETATM
2656
O
HOH
W
303
−17.878
33.786
16.873
1.00
50.15
O


HETATM
2657
O
HOH
W
304
−33.678
7.362
−8.182
1.00
49.39
O


HETATM
2658
O
HOH
W
305
−11.597
39.294
−8.859
1.00
53.94
O


HETATM
2659
O
HOH
W
306
2.882
29.239
3.191
1.00
45.89
O


HETATM
2660
O
HOH
W
307
−38.649
20.156
−7.082
1.00
49.36
O


HETATM
2661
O
HOH
W
308
−40.591
26.658
0.275
1.00
43.29
O


HETATM
2662
O
HOH
W
309
−6.537
4.311
13.825
1.00
47.40
O


HETATM
2663
O
HOH
W
310
−5.558
35.013
−13.517
1.00
48.80
O


HETATM
2664
O
HOH
W
311
−28.826
11.425
−15.322
1.00
49.38
O


HETATM
2665
O
HOH
W
312
−38.320
28.225
17.606
1.00
55.51
O


HETATM
2666
O
HOH
W
313
−23.593
4.828
13.752
1.00
27.36
O


HETATM
2667
O
HOH
W
314
−23.921
26.545
−16.243
1.00
34.27
O


HETATM
2668
O
HOH
W
315
−22.322
11.653
14.462
1.00
34.83
O


HETATM
2669
O
HOH
W
316
−12.651
34.045
−11.232
1.00
31.40
O


HETATM
2670
O
HOH
W
317
−11.298
29.991
11.215
1.00
37.94
O


HETATM
2671
O
HOH
W
318
−22.524
14.427
−23.824
1.00
34.38
O


HETATM
2672
O
HOH
W
319
−17.894
26.130
16.911
1.00
40.42
O


HETATM
2673
O
HOH
W
320
−30.095
30.303
9.084
1.00
43.47
O


HETATM
2674
O
HOH
W
321
−13.730
28.546
14.642
1.00
45.76
O


HETATM
2675
O
HOH
W
322
0.192
20.485
4.996
1.00
42.15
O


HETATM
2676
O
HOH
W
323
8.773
19.721
−2.795
1.00
41.00
O


HETATM
2677
O
HOH
W
324
−24.325
35.749
−12.402
1.00
41.84
O


HETATM
2678
O
HOH
W
325
−24.805
13.589
−22.690
1.00
42.74
O


HETATM
2679
O
HOH
W
326
−16.673
23.836
17.169
1.00
42.32
O


HETATM
2680
O
HOH
W
327
−26.618
15.414
−14.762
1.00
52.87
O


HETATM
2681
O
HOH
W
328
−17.876
41.703
11.298
1.00
40.21
O


HETATM
2682
O
HOH
W
329
2.710
12.479
−3.493
1.00
41.71
O


HETATM
2683
O
HOH
W
330
−40.522
22.831
11.667
1.00
40.68
O


HETATM
2684
O
HOH
W
331
−5.318
8.894
−11.975
1.00
45.24
O


HETATM
2685
O
HOH
W
332
6.465
34.844
−5.820
1.00
41.37
O


HETATM
2686
O
HOH
W
333
−7.294
7.847
−13.196
1.00
46.21
O


HETATM
2687
O
HOH
W
334
−27.575
−1.555
−9.482
1.00
43.31
O


HETATM
2688
O
HOH
W
335
−16.302
44.157
6.300
1.00
50.65
O


HETATM
2689
O
HOH
W
336
7.324
25.112
−0.042
1.00
46.30
O


HETATM
2690
O
HOH
W
337
−25.386
14.745
−27.433
1.00
48.90
O


HETATM
2691
O
HOH
W
338
−11.898
7.106
−19.877
1.00
48.25
O


HETATM
2692
O
HOH
W
339
−6.543
5.813
−9.614
1.00
50.85
O


HETATM
2693
O
HOH
W
340
−24.469
1.696
−2.733
1.00
49.67
O


HETATM
2694
O
HOH
W
341
−25.761
37.027
−10.672
1.00
43.85
O


HETATM
2695
O
HOH
W
342
−37.486
7.329
−1.731
1.00
48.83
O


HETATM
2696
O
HOH
W
343
−17.291
4.085
16.803
1.00
48.72
O


HETATM
2697
O
HOH
W
344
−14.036
26.435
−22.926
1.00
43.49
O


HETATM
2698
O
HOH
W
345
0.692
30.664
2.086
1.00
53.06
O


HETATM
2699
O
HOH
W
346
−41.805
14.047
−3.004
1.00
45.34
O


HETATM
2700
O
HOH
W
347
−5.313
18.738
12.526
1.00
56.10
O


HETATM
2701
O
HOH
W
348
−15.979
2.222
−18.286
1.00
52.97
O


HETATM
2702
O
HOH
W
349
12.845
27.550
2.215
1.00
51.89
O


HETATM
2703
O
HOH
W
350
−24.703
2.114
13.280
1.00
52.54
O


HETATM
2704
O
HOH
W
351
−13.629
8.385
17.755
1.00
53.31
O


HETATM
2705
O
HOH
W
352
0.600
10.756
1.065
1.00
52.20
O


HETATM
2706
O
HOH
W
353
−6.584
3.294
−8.664
1.00
58.25
O


HETATM
2707
O
HOH
W
354
−19.526
1.509
−15.069
1.00
58.64
O


HETATM
2708
O
HOH
W
355
−37.744
8.976
−6.842
1.00
49.38
O


HETATM
2709
O
HOH
W
356
11.571
31.022
2.199
1.00
49.80
O


HETATM
2710
O
HOH
W
357
−5.947
36.986
−4.732
1.00
50.31
O


HETATM
2711
O
HOH
W
358
−15.405
41.052
11.474
1.00
51.49
O


HETATM
2712
O
HOH
W
359
−12.833
40.449
−4.727
1.00
51.48
O


HETATM
2713
O
HOH
W
360
−5.438
1.935
−3.550
1.00
50.92
O


HETATM
2714
O
HOH
W
361
−10.249
0.894
−12.528
1.00
59.44
O


HETATM
2715
O
HOH
W
362
−31.963
40.849
5.308
1.00
56.17
O


HETATM
2716
O
HOH
W
363
−20.396
19.502
−28.110
1.00
57.97
O


HETATM
2717
O
HOH
W
364
−38.545
15.892
−6.873
1.00
53.86
O


HETATM
2718
O
HOH
W
365
−8.651
36.740
4.818
1.00
56.42
O


HETATM
2719
O
HOH
W
366
−16.900
45.179
−0.086
1.00
57.28
O


HETATM
2720
O
HOH
W
367
−15.600
42.705
−0.411
1.00
53.55
O


HETATM
2721
O
HOH
W
368
−1.122
22.193
−13.088
1.00
50.00
O


HETATM
2722
O
HOH
W
369
−3.015
2.725
5.944
1.00
49.17
O


HETATM
2723
O
HOH
W
370
−32.840
41.664
3.046
1.00
55.05
O


HETATM
2724
O
HOH
W
371
−31.332
6.082
11.720
1.00
55.05
O


HETATM
2725
O
HOH
W
372
−5.263
32.737
9.359
1.00
53.35
O


HETATM
2726
O
HOH
W
373
−17.185
11.511
17.118
1.00
55.25
O


HETATM
2727
O
HOH
W
374
−9.075
41.015
−9.468
1.00
55.28
O


HETATM
2728
O
HOH
W
375
−2.683
7.946
9.076
1.00
55.07
O


HETATM
2729
O
HOH
W
376
7.720
16.995
−3.341
1.00
52.87
O


HETATM
2730
O
HOH
W
377
−4.253
13.777
−12.953
1.00
51.97
O


END









REFERENCES
Patents


    U.S. Pat. No. 4,906,122A. Coupling for molecular models
    U.S. Pat. No. 5,030,103A. Dynamic molecular model
    U.S. Pat. No. 5,200,91lA. Method for modelling the electron density of a crystal
    U.S. Pat. No. 5,365,456A. Method for modelling the electron density of a crystal
    U.S. Pat. No. 5,583,973A. Molecular modeling method and system
    U.S. Pat. No. 5,612,894A. System and method for molecular modeling utilizing a sensitivity factor
    U.S. Pat. No. 5,733,720A. Genetically engineered cell lines for detecting infectious herpesvirus and methods therefor
    U.S. Pat. No. 5,763,263A. Method and apparatus for producing position addressable combinatorial libraries
    U.S. Pat. No. 5,942,428A. Crystals of the tyrosine kinase domain of non-insulin receptor tyrosine kinases
    U.S. Pat. No. 5,994,503A. Nucleotide and protein sequences of lats genes and methods based thereon
    U.S. Pat. No. 5,998,593A. Fluorescent enzyme substrates
    U.S. Pat. No. 6,037,117A. Methods using the Staphylococcus aureus glycyl tRNA synthetase crystalline structure
    U.S. Pat. No. 6,071,700A. Heterologous polypeptide production in the absence of nonsense-mediated mRNA decay functions
    U.S. Pat. No. 6,075,014A. Inhibitors of &bgr; -lactamases and uses therefor
    U.S. Pat. No. 6,075,123A. Cyclin-C variants, and diagnostic and therapeutic uses thereof
    U.S. Pat. No. 6,080,576A. Vectors for gene trapping and gene activation
    U.S. Pat. No. 6,093,573A. Three-dimensional structure of bactericidal/permeability-increasing protein (BPI)
    U.S. Pat. No. 6,172,262B1. Amphiphilic agents for membrane protein solubilization


Other References


    Adams, P. D., K. Gopal, et al. (2004). “Recent developments in the PHENIX software for automated crystallographic structure determination.” J Synchrotron Radiat 11 (Pt 1): 53-5.
    Adams, P. D., R. W. Grosse-Kunstleve, et al. (2002). “PHENIX: building new software for automated crystallographic structure determination.” Acta Crystallogr D Biol Crystallogr 58(Pt 11): 1948-54.
    Altschul, S. F. (1993). “A protein alignment scoring system sensitive at all evolutionary distances.” J. Mol. Evol. 36: 290-300.
    Altschul, S. F., M. S. Boguski, et al. (1994). “Issues in searching molecular sequence databases.” Nature Genetics 6: 119-129.
    Altschul, S. F., W. Gish, et al. (1990). “Basic local alignment search tool.” J Mol Biol 215(3): 403-10.
    Bacon, D. J. and J. Moult (1992). “Docking by Least-squares Fitting of Molecular Surface Patterns.” J. Mol. Biol. 225: 849-858.
    Bartlett, P. A., G. T. Shea, 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: 82-196.
    Bohm, H.-J. (1992). “The Computer Program LUDI: A New Method for the De Novo Design of Enzyme Inhibitors.” J. Computer-Aided Molecular Design 6: 61-78.
    Boutselakis, H., D. Dimitropoulos, et al. (2003). “E-MSD: the European Bioinformatics Institute Macromolecular Structure Database.” Nucleic Acids Res 31(1): 458-62.
    Bradford, M. M. (1976). “A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.” Anal Biochem 72: 248-54.
    Brady, L., A. M. Brzozowski, et al. (1990). “A serine protease triad forms the catalytic centre of a triacylglycerol lipase.” Nature 343(6260): 767-70.
    Brunger, A. T. (1993). X-Flor Version 3.1: A system for X-ray crystallography and NMR. New Haven, Conn., Yale Univ. Pr.
    Brzozowski, A. M., U. Derewenda, et al. (1991). “A model for interfacial activation in lipases from the structure of a fungal lipase-inhibitor complex.” Nature 351(6326): 491-4.
    Campbell (1984). Biological Spectroscopy. Menlo Park, Calif., The Benjamin/Cummings Publishing Co., Inc.
    Cantor, C. R. and P. R. Schimmel (1980). Biophysical Chemistry, Part II, “Techniques for the Study of Biological Structure and Function”, W.H. Freeman & Co.
    Carriere, F., K. Thirstrup, et al. (1997). “Pancreatic lipase structure-function relationships by domain exchange.” Biochemistry 36(1): 239-48.
    Chahinian, H., L. Nini, et al. (2002). “Distinction between esterases and lipases: a kinetic study with vinyl esters and TAG.” Lipids 37(7): 653-62.
    Cohen (Editor), N.C. (1996). Guidebook on Molecular Modeling in Drug Design, Academic Press.
    Cohen, N., J. Blaney, et al. (1990). “Molecular Modeling Software and Methods for Medicinal Chemistry.” J. Med. Chem. 33: 883-894.
    Crowther, J. R. (1995). ELISA: Theory and Practice (Methods in Molecular Biology), Humana Press.
    Delano, W. L. (2002). The PyMOL Molecular Graphics System. Palo Alto, Calif., USA.
    Devlin (Editor), J. P. (1998). In High Throughput Screening: The Discovery of Bioactive Substances. New York, Marcel Dekker Inc.
    Dinh, T. P., T. F. Freund, et al. (2002). “A role for monoglyceride lipase in 2-arachidonoylglycerol inactivation.” Chem Phys Lipids 121(1-2): 149-58.
    Dinh, T. P., S. Kathuria, et al. (2004). “RNA interference suggests a primary role for monoacylglycerol lipase in the degradation of the endocannabinoid 2-arachidonoylglycerol.” Mol Pharmacol 66(5): 1260-4.
    Drenth, J. (1999). Principles of Protein X-ray Crystallography (Springer Advanced Texts in Chemistry). Berlin, Springer Verlag.
    Dugi, K. A., H. L. Dichek, et al. (1995). “Human hepatic and lipoprotein lipase: the loop covering the catalytic site mediates lipase substrate specificity.” J Biol Chem 270(43): 25396-401.
    Dugi, K. A., H. L. Dichek, et al. (1992). “Human lipoprotein lipase: the loop covering the catalytic site is essential for interaction with lipid substrates.” J Biol Chem 267(35): 25086-91.
    Emsley, P. and K. Cowtan (2004). “Coot: model-building tools for molecular graphics.” Acta Crystallogr D Biol Crystallogr 60(Pt 12 Pt 1): 2126-32.
    Farquhar-Smith, W. P., M. Egertova, et al. (2000). “Cannabinoid CB(1) receptor expression in rat spinal cord.” Mol Cell Neurosci 15(6): 510-21.
    Faustinella, F., L. C. Smith, et al. (1992). “Functional topology of a surface loop shielding the catalytic center in lipoprotein lipase.” Biochemistry 31(32): 7219-23.
    Frisch, M. J., G. W. Trucks, et al. (1992). “Gaussian 92, Revision C.” Gaussian, Inc. 
    Galiegue, S., S. Mary, et al. (1995). “Expression of central and peripheral cannabinoid receptors in human immune tissues and leukocyte subpopulations.” Eur J Biochem 232(1): 54-61.
    Gans, W., A. Amann, et al. (1996). Fundamental Principals of Molecular Modeling, Plenum Pub. Corp.
    Gonsiorek, W., C. Lunn, et al. (2000). “Endocannabinoid 2-arachidonyl glycerol is a full agonist through human type 2 cannabinoid receptor: antagonism by anandamide.” Mol Pharmacol 57(5): 1045-50.
    Goodford, P. J. (1985). “A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules.” J. Med. Chem. 28: 849-857.
    Goodsell, D. S, and A. J. Olsen (1990). “Automated Docking of Substrates to Proteins by Simulated Annealing.” Proteins: Structure. Function, and Genetics 8: 195-202.
    Guindon, J., J. Desroches, et al. (2007). “The antinociceptive effects of intraplantar injections of 2-arachidonoyl glycerol are mediated by cannabinoid CB2 receptors.” Br J Pharmacol 150(6): 693-701.
    Hanus, L., A. Breuer, et al. (1999). “HU-308: a specific agonist for CB(2), a peripheral cannabinoid receptor.” Proc Natl Acad Sci USA 96(25): 14228-33.
    Heikinheimo, P., A. Goldman, et al. (1999). “Of barn owls and bankers: a lush variety of alpha/beta hydrolases.” Structure 7(6): R141-6.
    Henikoff, J. G. (1992). “Amino acid substitution matrices from protein blocks.” Proc. Natl. Acad. Sci. USA (89): 10915-10919.
    Higgins, D. G., J. D. Thompson, et al. (1996). “Using CLUSTAL for multiple sequence alignments.” Methods Enzymol 266: 383-402.
    Hohmann, A. G. (2002). “Spinal and peripheral mechanisms of cannabinoid antinociception: behavioral, neurophysiological and neuroanatomical perspectives.” Chem Phys Lipids 121(1-2): 173-90.
    Hohmann, A. G., J. N. Farthing, et al. (2004). “Selective activation of cannabinoid CB2 receptors suppresses hyperalgesia evoked by intradermal capsaicin.” J Pharmacol Exp Ther 308(2): 446-53.
    Hohmann, A. G. and M. Herkenham (1999). “Cannabinoid receptors undergo axonal flow in sensory nerves.” Neuroscience 92(4): 1171-5.
    Hohmann, A. G., R. L. Suplita, et al. (2005). “An endocannabinoid mechanism for stress-induced analgesia.” Nature 435(7045): 1108-12.
    Holmquist, M. (2000). “Alpha/Beta-hydrolase fold enzymes: structures, functions and mechanisms.” Curr Protein Pept Sci 1(2): 209-35.
    Ibrahim, M. M., M. L. Rude, et al. (2006). “CB2 cannabinoid receptor mediation of antinociception.” Pain 122(1-2): 36-42.
    Ishikawa, E. (1999). Ultrasensitive and rapid enzyme immunoassay, In: Laboratory Techniques in Biochemistry and Molecular Biology. Amsterdam, Elsevier.
    Kaneko, T., N. Tanaka, et al. (2005). “Crystal structures of RsbQ, a stress-response regulator in Bacillus subtilis.” Protein Sci 14(2): 558-65.
    Karlin, S, and S. F. Altschul (1990). “Methods for assessing the statistical significance of molecular sequence features by using general scoring schemes.” Proc. Natl. Acad. Sci. USA 87: 2264-2268.
    Karlsson, M., J. A. Contreras, et al. (1997). “cDNA cloning, tissue distribution, and identification of the catalytic triad of monoglyceride lipase. Evolutionary relationship to esterases, lysophospholipases, and haloperoxidases.” J Biol Chem 272(43): 27218-23.
    Karlsson, M., K. Reue, et al. (2001). “Exon-intron organization and chromosomal localization of the mouse monoglyceride lipase gene.” Gene 272(1-2): 11-8.
    Karlsson, M., H. Tornqvist, et al. (2000). “Expression, purification, and characterization of histidine-tagged mouse monoglyceride lipase from baculovirus-infected insect cells.” Protein Expr Purif 18(3): 286-92.
    Kemeny (Editor), D. M. and S. J. Challacombe (Editor) (1988). Elisa and Other Solid Phase Immunoassays Theoretical and Practical Aspects, New York, John Wiley and Sons.
    Kemeny, D. M. (1991). A Practical Guide to ELISA, Pergamon Press.
    Kuntz, I. D., J. M. Blaney, et al. (1982). “A geometric approach to macromolecule-ligand interactions.” J Mol Biol 161(2): 269-88.
    Lee, C. and K. Irizarry (2001). “The GeneMine system for genome/proteome annotation and collaborative data-mining.” IBM Systems Journal 40(2).
    Levitt, M. (1992). “Accurate modeling of protein conformation by automatic segment matching.” J Mol Biol 226(2): 507-33.
    Lipinski, C., F. Lombardo, et al. (1997). “Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings” Advanced Drug Delivery Reviews 23(1-3): 3-25.
    Longenecker, K. L., S. M. Garrard, et al. (2001). “Protein crystallization by rational mutagenesis of surface residues: Lys to Ala mutations promote crystallization of RhoGDI.” Acta Crystallogr D Biol Crystallogr 57(Pt 5): 679-88.
    Makara, J. K., M. Mor, et al. (2005). “Selective inhibition of 2-AG hydrolysis enhances endocannabinoid signaling in hippocampus.” Nat Neurosci 8(9): 1139-41.
    Malan, T. P., Jr., M. M. Ibrahim, et al. (2001). “CB2 cannabinoid receptor-mediated peripheral antinociception.” Pain 93(3): 239-45.
    Malan, T. P., Jr., M. M. Ibrahim, et al. (2002). “Inhibition of pain responses by activation of CB(2) cannabinoid receptors.” Chem Phys Lipids 121(1-2): 191-200.
    Maresz, K., E. J. Carrier, et al. (2005). “Modulation of the cannabinoid CB2 receptor in microglial cells in response to inflammatory stimuli.” J Neurochem 95(2): 437-45.
    Martin, Y. C. (1992). “3D Database Searching in Drug Design.” J. Med. Chem. 35: 2145-2154.
    Mary Ann Liebert (Publishers), I. (1995). The BIOTECHNOLOGY SOFTWARE DIRECTORY, A Buyer's Guide. Larchmont, N.Y. Mary Ann Liebert, Inc., Publishers.
    Mateja, A., Y. Devedjiev, et al. (2002). “The impact of Glu→Ala and Glu→Asp mutations on the crystallization properties of RhoGDI: the structure of RhoGDI at 1.3 A resolution.” Acta Crystallogr D Biol Crystallogr 58(Pt 12): 1983-91.
    Matsuda, L. A., S. J. Lolait, et al. (1990). “Structure of a cannabinoid receptor and functional expression of the cloned cDNA.” Nature 346(6284): 561-4.
    Matteucci and J. Caruthers (1981). J. Am. Chem. Soc. 103(3): 185-3191.
    Mccoy, A. J., R. W. Grosse-Kunstleve, et al. (2007). “Phaser crystallographic software.” Journal of Applied Crystallography 40(4): 658-674.
    Meng, E. C., B. K. Shoichet, et al. (1992). “Automated docking with grid-based energy evaluation.” J. Comp. Chem. 13: 505-524.
    Miranker, A. and M. Karplus (1991). “Functionality Maps of Binding Sites: A Multiple Copy Simultaneous Search Method.” Proteins: Structure, Function and Genetics 11: 29-34.
    Munro, S., K. L. Thomas, et al. (1993). “Molecular characterization of a peripheral receptor for cannabinoids.” Nature 365(6441): 61-5.
    Navia, M. A. and M. A. Murcko (1992). “The Use of Structural Information in Drug Design.” Current Opinions in Structural Biology 2: 202-210
    Nini, L., L. Sarda, et al. (2001). “Lipase-catalysed hydrolysis of short-chain substrates in solution and in emulsion: a kinetic study.” Biochim Biophys Acta 1534(1): 34-44.
    Nishibata, Y. and A. Itai (1991). “Automatic creation of drug candidate structures based on receptor structure. Starting point for artificial lead generation.” Tetrahedron 47: 8985-8990.
    Norton, P. A. and J. M. Coffin (1985). “Bacterial beta-galactosidase as a marker of Rous sarcoma virus gene expression and replication.” Mol Cell Biol 5(2): 281-90.
    Ollis, D. L., E. Cheah, et al. (1992). “The alpha/beta hydrolase fold.” Protein Eng 5(3): 197-211.
    Otwinowski, Z. and W. Minor (1997). Processing of X-ray Diffraction Data Collected in Oscillation Mode. Methods in Enzymology. C. W. Carter and R. M. Sweet. New York, Academic Press. 276: 307-326.
    Pantoliano, M. W., E. C. Petrella, et al. (2001). “High-density miniaturized thermal shift assays as a general strategy for drug discovery.” J Biomol Screen 6(6): 429-40.
    Quartilho, A., H. P. Mata, et al. (2003). “Inhibition of inflammatory hyperalgesia by activation of peripheral CB2 cannabinoid receptors.” Anesthesiology 99(4): 955-60.
    Rice, A. S., W. P. Farquhar-Smith, et al. (2002). “Endocannabinoids and pain: spinal and peripheral analgesia in inflammation and neuropathy.” Prostaglandins Leukot Essent Fatty Acids 66(2-3): 243-56.
    Richardson, J. D. (2000). “Cannabinoids modulate pain by multiple mechanisms of actions.” The Journal of Pain 1(1): 2-14.
    Richardson, J. D., S. Kilo, et al. (1998). “Cannabinoids reduce hyperalgesia and inflammation via interaction with peripheral CB1 receptors.” Pain 75(1): 111-9.
    Rossmann, M. G. (1972). The molecular replacement method; a collection of papers on the use of non-crystallographic symmetry., Gordon & Breach, New York.
    Rotstein, S. H. and M. A. Murcko (1993). “GroupBuild: a fragment-based method for de novo drug design.” J Med Chem 36(12): 1700-10.
    Roussel, A., S. Canaan, et al. (1999). “Crystal structure of human gastric lipase and model of lysosomal acid lipase, two lipolytic enzymes of medical interest.” J Biol Chem 274(24): 16995-7002.
    Roussel, A., N. Miled, et al. (2002). “Crystal structure of the open form of dog gastric lipase in complex with a phosphonate inhibitor.” J Biol Chem 277(3): 2266-74.
    Saario, S. M., A. Poso, et al. (2006). “Fatty acid amide hydrolase inhibitors from virtual screening of the endocannabinoid system.” J Med Chem 49(15): 4650-6.
    Saario, S. M., O. M. Salo, et al. (2005). “Characterization of the sulfhydryl-sensitive site in the enzyme responsible for hydrolysis of 2-arachidonoyl-glycerol in rat cerebellar membranes.” Chem Biol 12(6): 649-56.
    Saario, S. M., J. R. Savinainen, et al. (2004). “Monoglyceride lipase-like enzymatic activity is responsible for hydrolysis of 2-arachidonoylglycerol in rat cerebellar membranes.” Biochem Pharmacol 67(7): 1381-7.
    Sambrook, J., E. F. Fritsch, et al. (1989). Molecular cloning. 2nd ed. New York: Cold Spring Harbor Laboratory Press.
    Schlecht, M. (1998). Molecular Modeling on the PC, John Wiley & Sons.
    Schrag, J. D., Y. G. Li, et al. (1991). “Ser-His-Glu triad forms the catalytic site of the lipase from Geotrichum candidum.” Nature 351(6329): 761-4.
    Segel, I. H. (1975). Enzyme Kinetics: Behavior and Analysis of Rapid Equilibrium and Steady-State Enzyme Systems, J. Willey & Sons.
    Smith, W. B. (1996). Introduction to Theoretical Organic. Chemistry and Molecular Modeling. New York, VCH Publishers.
    Somma-Delpero, C., A. Valette, et al. (1995). “Purification and properties of a monoacylglycerol lipase in human erythrocytes.” Biochem J 312 (Pt 2): 519-25.
    Sugiura, T., S. Kondo, et al. (2000). “Evidence that 2-arachidonoylglycerol but not N-palmitoylethanolamine or anandamide is the physiological ligand for the cannabinoid CB2 receptor. Comparison of the agonistic activities of various cannabinoid receptor ligands in HL-60 cells.” J Biol Chem 275(1): 605-12.
    Sussman, J. L., D. Lin, et al. (1998). “Protein Data Bank (PDB): database of three-dimensional structural information of biological macromolecules.” Acta Crystallogr D Biol Crystallogr 54(Pt 6 Pt 1): 1078-84.
    Tatusova, T. A. and T. L. Madden (1999). “BLAST 2 Sequences, a new tool for comparing protein and nucleotide sequences.” FEMS Microbiol Lett 174(2): 247-50.
    Terwilliger, T. C., R. W. Grosse-Kunstleve, et al. (2008). “Iterative model building, structure refinement and density modification with the PHENIX AutoBuild wizard.” Acta Crystallogr D Biol Crystallogr 64(Pt 1): 61-9.
    Thompson, J. D., D. G. Higgins, et al. (1994). “CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.” Nucleic Acids Res 22(22): 4673-80.
    Tornqvist, H. and P. Belfrage (1976). “Purification and some properties of a monoacylglycerol-hydrolyzing enzyme of rat adipose tissue.” J Biol Chem 251(3): 813-9.
    Travis, J. (1993). “Proteins and Organic Solvents Make an Eye-Opening Mix.” Science 262:1374
    Tsirelson, V. G. and R. P. Ozerov (1996). Electron Density and Bonding in Crystals: Principles, Theory and X-ray Diffraction Experiments in Solid State Physics and Chemistry, Inst. of Physics Pub.
    Van Den Berg, B., M. Tessari, et al. (1995). “NMR structures of phospholipase A2 reveal conformational changes during interfacial activation.” Nat Struct Biol 2(5): 402-6.
    Van Den Berg, B., M. Tessari, et al. (1995). “Solution structure of porcine pancreatic phospholipase A2 complexed with micelles and a competitive inhibitor.” J Biomol NMR 5(2): 110-21.
    Van Tilbeurgh, H., L. Sarda, et al. (1992). “Structure of the pancreatic lipase-procolipase complex.” Nature 359(6391): 159-62.
    Vandevoorde, S, and D. M. Lambert (2005). “Focus on the three key enzymes hydrolysing endocannabinoids as new drug targets.” Curr Pharm Des 11(20): 2647-68.
    Walczak, J. S., V. Pichette, et al. (2005). “Behavioral, pharmacological and molecular characterization of the saphenous nerve partial ligation: a new model of neuropathic pain.” Neuroscience 132(4): 1093-102.
    Wall, E. M., J. Cao, et al. (1997). “A novel poxvirus gene and its human homolog are similar to an E. coli lysophospholipase.” Virus Res 52(2): 157-67.
    Wang, X., C. S. Wang, et al. (1997). “The crystal structure of bovine bile salt activated lipase: insights into the bile salt activation mechanism.” Structure 5(9): 1209-18.
    Winkler, F. K., A. D'arcy, et al. (1990). “Structure of human pancreatic lipase.” Nature 343(6260): 771-4.
    Woolfson, M. M. (1997). An Introduction to X-ray Crystallography. Cambridge, UK, Cambridge Univ. Pr.

## Claims

1. The A crystal comprising monoacylglycerol lipase (MGLL) peptide consisting of SEQ ID NO: 7 and a ligand, wherein said ligand is selected from the group consisting of the following structures:




and wherein said crystal has a space group of C2221; and a unit cell having dimensions of a=93.95 Å, b=128.45 Å, c=60.6 Å.

2. A method for the production of a crystal complex comprising a monoacylglycerol lipase (MGLL) peptide consisting of SEQ ID NO: 7 with a ligand of the following structure:




comprising:
a. contacting the MGLL polypeptide with said ligand in a suitable solution comprising 8% PEG MME 5K, 100 mM Na Citrate pH5.5 and 2% n-Octyl-Beta-D-Glucopyranoside, and,
b. crystallizing said resulting complex of MGLL polypeptide-ligand from said solution.