# Patent

## Title
Three-dimensional structure of chalcone isomerase and methods of use thereof

## Abstract

This disclosure provides crystalline flavonoid or flavanone isomerases, isolated non-native isomerase having the structural coordinates of said crystalline isomerase, and nucleic acids encoding such non-native isomerase. Also disclosed are methods of predicting the activity and/or substrate specificity of a putative isomerase, methods of identifying potential isomerase substrates, and methods of identifying potential isomerase inhibitors.

## Background
RELATED APPLICATIONS
This application claims the benefit of U.S. Application No. 60/229,277, filed Aug. 30, 2000, which is hereby incorporated by reference herein in its entirety.


STATEMENT OF GOVERNMENT SUPPORT
This work is supported in part by grant number MCB-9982586 from the National Science Foundation. The Government has certain rights to this invention.
ACKNOWLEDGMENT
This invention was made with United States Government support under Grant No. MCB-9982586, awarded by the National Science Foundation. The Government has certain rights in the invention.


FIELD OF THE INVENTION
The present invention relates to methods for designing mutant chalcone isomerases, and to predicting the activity and/or substrate specificity of native and mutant chalcone isomerases. The present invention further relates to methods for identifying chalcone isomerase substrates and/or inhibitors.
BACKGROUND
Advances in molecular biology have allowed the development of biological agents useful in modulating protein or nucleic acid activity or expression, respectively. Many of these advances are based on identifying the primary sequence of the molecule to be modulated. For example, determining the nucleic acid sequence of DNA or RNA allows the development of antisense or ribozyme molecules. Similarly, identifying the primary sequence allows for the identification of sequences that may be useful in creating monoclonal antibodies. However, often the primary sequence of a protein is insufficient to develop therapeutic or diagnostic molecules due to the secondary, tertiary or quartenary structure of the protein from which the primary sequence is obtained. The process of designing potent and specific inhibitors, activators, or novel proteins has improved with the arrival of techniques for determining the three-dimensional structure of an enzyme or polypeptide whose activity one desires to modulate.
The phenylpropanoid synthetic pathway in plants produces a class of compounds know as anthocyanins, which are used for a variety of applications. Anthocyanins are involved in pigmentation and protection against UV photodamage, synthesis of anti-microbial phytoalexins, and are flavonoid inducers of Rhizobium modulation genes 1-4. As medicinal natural products, the phenylpropanoids exhibit cancer chemopreventive activity, as well as anti-mitotic, estrogenic, anti-malarial, anti-oxidant, and antiasthmatic activities. The benefits of consuming, red wine, which contains significant amounts of 3,4′,5-trihydroxystilbene (resveratrol) and other phenylpropanoids, highlight the dietary importance of these compounds. One strategy for the generation of novel enzymatic activity in flavonoid biosynthesis uses protein-engineering methods and requires a detailed structural knowledge of enzymes within the targeted pathway.
Polyketides are a large class of compounds and include a broad range of antibiotics, immunosuppressants and anticancer agents which together account for sales of over $5 billion per year. Polyketides are molecules which are an extremely rich source of bioactivities, including antibiotics (e.g., tetracyclines and erythromycin), anti-cancer agents (e.g., daunomycin), immunosuppressants (e.g., FK506 and rapamycin), veterinary products (e.g., monensin), and the like. Many polyketides (produced by polyketide synthases) are valuable as therapeutic agents. Polyketide synthases are multifunctional enzymes that catalyze the biosynthesis of a huge variety of carbon chains differing in length and patterns of functionality and cyclization.
Chalcone synthase (CHS), a polyketide synthase, plays an essential role in the biosynthesis of plant phenylpropanoids. CHS supplies 4,2′,4′,6′-tetrahydroxychalcone (chalcone) to downstream enzymes, such as chalcone isomerase (CHI), that synthesize a diverse set of flavonoid phytoalexins and anthocyanin pigments.
An improvement in the understanding of the structure/function of these enzymes would allow for a number of advances in the art, e.g., the exploitation of the synthetic capabilities of known enzymes for production of useful new chemical compounds, for the creation of novel non-native enzymes having new synthetic capabilities etc. A need exists, therefore, for a detailed understanding of the molecular basis of the chemical reactions involved in polyketide, flavanone and flavonoid synthesis. The present invention addresses this and related needs.
SUMMARY OF THE INVENTION
In accordance with the present invention there are presented crystalline chalcone isomerases (CHIs) and the three-dimensional coordinates derived therefrom. Three-dimensional coordinates have been obtained for an active form of chalcone isomerase and the active site thereof, both with and without product or product analog. Accordingly, the three-dimensional coordinates and crystal structure of a CHI provides the ability to develop novel substrates, proteins and enzymatic products of CHI. In addition, the invention provides the use of the three-dimensional structure either alone or together with the structure of polyketide synthases, such as chalcone synthase, to provide a useful template for engineering novel enzymes and enzyme pathways to diversify and modify flavonoid biosynthesis for crop and food sources, as well as providing novel flavanones for intermediates and leads in drug discovery (see WO/01/07579 A2, published Feb. 1, 2001, the disclosure of which is incorporated herein by reference in it entirety).
One aspect of the present invention made possible by the results described herein is a model of the three-dimensional properties of chalcone isomerase proteins. In particular, the invention provides the three-dimensional properties of the active site. The invention features specific coordinates of at least twelve α-carbon atoms defining the active site in three-dimensional space. R-groups attached to said α-carbons are defined such that mutants can be made by changing at least one R-group found in the isomerase active site. Such mutants have unique and useful properties. Thus, in accordance with another embodiment of the invention, there are provided isolated non-native (e.g., mutant) isomerase(s) having at least twelve active site α-carbons having the structural coordinates disclosed herein and one or more R-groups other than those found in native chalcone isomerase(s).
The chalcone isomerase used in the crystallization studies disclosed herein is a chalcone isomerase derived from Medicago sativa (alfalfa) (SEQ ID NO:1). A large number of isomerase proteins from various plant species have primary amino acid sequences showing substantial homology and conservation. Thus, the three-dimensional coordinates disclosed herein can be employed in a variety of methods extending to various isomerase proteins. Accordingly, in another embodiment of the present invention, there are provided methods for predicting the activity and/or substrate specificity of a putative chalcone isomerase from a variety of species. There are further provided methods for identifying potential substrates for a chalcone isomerase, as well as inhibitors thereof.
Other aspects, embodiments, advantages, and features of the present invention will become apparent from the following specification.



BRIEF DESCRIPTION OF FIGURES
FIG. 1A shows the overall reaction catalyzed by CHI, which involves a Michael-type nucleophilic attack of the 2′-hydroxyl on the α,β-unsaturated double bond of chalcone. The numbering systems for chalcones (left) and flavanones (right) are shown.
FIG. 1B is a schematic ribbon diagram of the overall structure of chalcone isomerase (CHI). The N- and C-termini are labeled, as are the β-strands and α-helices of the structure. The position of (2S)-naringenin is also indicated.
FIG. 1C is a stereo-view of the Cα-backbone. This orientation is the same as in (FIG. 1A). Every tenth residue is numbered. The position of (2S)-naringenin is also shown.
FIG. 1D is a primary and secondary structure of CHI from Medicago sativa (alfalfa; P28012; see also SEQ ID NO:1, wherein the amino acid at residue 36 is Arg, the amino acid at residue 38 is Leu, the amino acid at residue 48 is Thr, the amino acid at residue 106 is Tyr, the amino acid at residue 109 is Lys, the amino acid at residue 110 is Val, the amino acid at residue 113 is Asn, the amino acid at residue 190 is Thr, and the amino acid at residue 191 is Met) and sequence alignment of CHIs from Phaseolus vulgaris (bean; P14298; SEQ ID NO:2), Pisum sativum (pea; P41089; SEQ ID NO:3), Zea maize (corn; S41579; SEQ ID NO:4), Vitis vinifera (grape; P51117; SEQ ID NO:5), Ipomoea purpurea (morning glory; af028238; SEQ ID NO:6), Petunia hybrida (P11651; SEQ ID NO:7), and Arabidopsis thaliana (P41088; SEQ ID NO:8). α-Helices and β-strands of CHI are indicated above the sequence and the numbering of each protein in parenthesis. Every tenth position in the alignment is dotted. Residues of the (2S)-naringenin binding cleft (underlined), residues of the active site hydrogen bond network (boxed), and other conserved residues (shaded) are indicated as noted. The residues that may influence substrate preference between chalcone and 6′-deoxychalcone are indicated with an asterisk.
FIG. 2 collectively shows 2 (2S)-Naringenin binding and structure of the active site cleft.
FIG. 2A shows a stereo-view of the SIGMAA-weighted |2Fo-Fc| electron density (1.2σ) for (2S)-naringenin.
FIG. 2B shows a stereo-view of residues in the active site cleft. (2S)-Naringenin and a water molecule are also shown. Hydrogen bond interactions are indicated with dotted lines. This view is oriented looking into the cleft. The surface corresponding to Lys 109 and Asn 113 (SEQ ID NO:9) was removed for clarity.
FIG. 2C shows a stereo-view surface representation of active site cleft showing the fit of (2S)-naringenin and a water molecule therein.
FIG. 3 collectively shows the proposed enzyme-mediated stereochemical control of the cyclization reaction. The surface of the binding deft is transparent to show selected residues (grey). The surface associated with Lys 109 and Asn 113 has been removed for clarity. The position of chalcone (hatched) prior to cyclization has been modeled to show the formation of the new bond (dotted line). Hydrogen bond interactions are indicated by dotted lines.
FIG. 3A shows a stereo-view of the proposed chalcone conformation leading to (2S)-naringenin formation.
FIG. 3B shows a stereo-view of the steric clash that prevents (2R)-naringenin formation.
FIG. 4 collectively shows the proposed reaction mechanism of CHI.
FIG. 4A is a view of the active site hydrogen bond network. This view is oriented looking out of the active site cleft. Dotted lines indicate hydrogen bonds.
FIG. 4B is a schematic representation of the active site hydrogen bonds. Distances are indicated in Å.
FIG. 4C is a proposed cyclization reaction catalyzed by CHI. Following nucleophilic attack of the 2′-O— of the substrate on the double bond in a Michael addition, the water molecule stabilized by Tyr 106 (see SEQ ID NO:1) acts as a general acid to stabilize the enolate. This results in formation of a flav-3-en-4-ol intermediate that tautomerizes into the reaction product.
FIG. 5 shows an example of a computer system in block diagram form.



DETAILED DESCRIPTION OF THE INVENTION
In flavonoid biosynthesis, chalcone isomerase (CHI, E.C. 5.5.1.6) catalyzes the cyclization of chalcone (4,2′,4′,6′-tetrahydroxychalcone) and 6′-deoxychalcone (4,2′,4′-trihydroxychalcone) into (2S)-naringenin (5,7,4′-trihydroxyflavanone) and (2S)-5-deoxyflavanone (7,4′-dihydroxyflavanone), respectively. Since chalcone spontaneously cyclizes into (2S/2R)-naringenin, CHI guarantees formation of the biologically active (S)-isomer. (2S)-Naringenin is the precursor of anthocyanin pigments, and mutations in the gene encoding CHI are linked to changes in floral pigmentation. (2S)-Naringenin and other flavonoids also act as small molecule transcription activators that target bacterial transcription regulators governing expression of Rhizobium genes involved in root nodulation.
An “isomerase” or a “chalcone isomerase” includes any one of a family of isomerase enzymes that catalyze the formation of flavonoid or flavanone compounds. Chalcone isomerases are generally monomers.
Mechanistically, CHI catalyzes the cyclization of chalcone with an apparent 100,000:1 preference for the S-isomer over the R-isomer. The second-order rate constant (kcat/Km) for conversion of chalcone by CHI approaches the diffusion-controlled limit with an enzyme-catalyzed rate that exceeds the spontaneous conversion rate by 107-fold. Combined with structural knowledge, the comparison of the spontaneous and enzyme-catalyzed reactions provides insight on how an enzyme accelerates the rate of an intramolecular chemical reaction. The present invention provides a 2.5 Å crystal structure of CHI from Medicago sativa (alfalfa) (SEQ ID NO:1) by multiple isomorphous replacement with anomalous scattering (MIRAS) and the 1.85 Å resolution structure of CHI complexed with (2S)-naringenin by difference Fourier analysis. Atomic resolution structures provide a molecular understanding of how CHI recognizes and catalyzes the stereospecific cyclization of chalcone and provides the ability to modulate natural product specificity and to develop novel isomerase proteins having substrate specificities.
CHI is a functional monomer of approximately 220 residues and has been isolated from a variety of higher plants. (Bednar, R. A. & Hadcock, J. R. J. Biol. Chem. 263:9582-9588, 1988; Dixon, et al. Phytochemistry 27:2801-2808, 1988). The present invention provides the first crystal structure for chalcone isomerase, which resembles an upside-down bouquet that adopts an open-faced β-sandwich fold (FIGS. 1B and 1C). A large β-sheet (β3a-β3f) and a layer of α-helices (α1-α7) comprise the core structure with three short β-strands (β1a, β1b, β2) on the opposite side of the large β-sheet. A search of the Protein Data Bank using DALI (Sander, C. & Schneider, R. Proteins Struct. Funct. Genet. 9:56-68, 1991) revealed no other structurally homologous folds. In addition, a PSI-BLAST search of sequence databases showed that CHI-like sequences are currently found in a number of plants and these sequences display limited homology with other proteins. These results imply that the CHI three-dimensional fold and enzymatic activity are found typically in the plant kingdom. Amino acid sequence comparison of CHIs from a variety of advanced land plants (SEQ ID NOs:1-8) reveals high homology (49% to 82% amino acid sequence identity) with regions of conservation spread uniformly throughout the primary structure (see FIG. 1D). A conservation of residues spanning β3a, β3b, α4, and α6 in the three-dimensional structure among CHIs from different species is demonstrated by the present invention as structural elements of the active site on the protein surface.
The data demonstrates that co-localization of proteins in loosely associated macromolecular complexes is a fundamental component of cellular processes, including flavonoid biosynthesis. CHI and other flavonoid biosynthetic enzymes may associate to provide efficient channeling of substrates and products as shown recently in Arabidopsis thaliana. Although the three short β-strands (β1a, β1b, β2) on the backside of the CHI structure (the relevant portion of the CHI structure is presented in SEQ ID NO:9) form a relatively flat surface that would be ideal for protein-protein interactions, both gel filtration and analytical ultracentrifugation failed to detect association of alfalfa CHI (SEQ ID NO:1) and alfalfa chalcone synthase 2 in vitro.
For the first time the present invention identifies the active site of CHI by identifying the location of (2S)-naringenin in the CHI structure (see SEQ ID NO:9 and FIG. 2). Although a commercially obtained mixture of (2S)- and (2R)-naringenin was used for co-crystallization, only the (2S)-isomer bound the CHI active site. The position of the (2S)-naringenin binding cleft is consistent with inactivation studies that suggested a cysteine residue (Cys 114 in alfalfa CHI, SEQ ID NO:1) is proximal to the active site (Bednar et al. J. Biol. Chem. 264:14272-14276, 1989). In the CHI structure, Cys 114 is near the binding cleft but does not directly contact (2S)-naringenin. The active site cleft is largely apolar and consists of residues from β3a (Arg 36, Gly 37, Leu 38), β3b (Phe 47, Thr 48, Ile 50), α4 (Tyr 106, Lys 109, Val 110, Asn 113), and α6 (Thr 190, Met 191) (FIG. 2B, SEQ ID NO:9). In addition, residues Ala 49, Lys 97, Leu 101, Glu 105, Glu 112, Cys 114, Tyr 152, Val 187, Asp 200 and Leu 201 (see SEQ ID NO:9) contact the ligand (naringenin or deoxyflavanones) or butress the above residues of β3a, β3b, α4, or α6. The apolar methylene carbons of Arg 36 are positioned by a restraining charge-charge interaction from the δ-guanido group to Glu 200 (see SEQ ID NO:9). In addition, the methylene carbons of Lys 109 are fixed by a charge-charge interaction between the side chain amine and Glu 112. Except for Thr 190 and Met 191, the residues contacting (2S)-naringenin are identical among CHIs from different plants (FIG. 1D, SEQ ID NOs:1-8). Although van der Waals contacts dominate the interactions between CHI and (2S)-naringenin, two hydrogen bond interactions exist. The first is mediated by the side chain hydroxyl moiety of Thr 48 bound to the 4′-hydroxyl group of (2S)-naringenin. The second interaction is between a water molecule and the ligand ketone (FIG. 2B). This water molecule and its connected network of hydrogen bonds occupy the same position in the apoenzyme structure. The overall surface topology of the cleft tightly sequesters the (2S)-naringenin molecule (FIG. 2C). The CHI•naringenin complex (SEQ ID NO:9) explains the stereochemical preference of the cyclization reaction; moreover, it suggests why CHIs from different species show moderate selectivity for chalcone and 6′-deoxychalcone as substrates.
“Active Site” refers to a site in an isomerase defined by amino acid residues that interact with substrate and facilitate a biosynthetic reaction that allows one or more products to be produced. An active site is comprised of α-carbon atoms that are indirectly linked via peptide bonds and have the structural coordinates disclosed by the atoms of the residues found in the β3a, β3b, α4 and α6 regions of chalcone isomerase (e.g., Arg 36, Gly 37, Leu 38, Phe 47, Thr 48, Ile 50, Tyr 106, Lys 109, Val 110, Asn 113, Thr 190, and Met 191 of SEQ ID NO:1). In addition, residues Ala 49, Lys 97, Leu 101, Glu 105, Glu 112, Cys 114, Tyr 152, Val 187, Asp 200 and Leu 201 (see SEQ ID NO:1) contact the ligand (naringenin or deoxyflavanones) or butress the above residues of β3a, β3b, α4, or α6. The position in three-dimensional space of an α-carbon at the active site of an isomerase and of R-groups associated therewith can be determined using techniques such as three-dimensional modeling, X-ray crystallography, and/or techniques associated therewith.
Modeling of chalcone, based on the position of (2S)-naringenin, shows that a slight rotation of the trihydroxyl-ring outward in the direction of the active site opening places the 2′-hydroxyl group in position for nucleophilic attack on the α,β-unsaturated double bond of the coumaroyl moiety (FIG. 3A) This rotation preserves the position of the chalcone backbone and the hydrogen bonds between Thr 190 and the water molecule at the backside of the binding site. Formation of (2R)-naringenin would require substantial rearrangements in the active site of CHI due to significant steric clashes between the trihydroxyl-ring and CHI side chains. Although rotation of the trihydroxyl-ring away from the active site entrance could reposition the 2′-hydroxyl group for attack on the opposite face of the α,β-double bond, the side chain of Val 110 sterically prevents this movement from occurring (FIG. 3B). In addition, the opposite side of the substrate double bond could be positioned for attack by the 2′-hydroxyl group in the formation of (2R)-naringenin. This alternative cyclization would be accomplished by rotation of the coumaroyl moiety outward towards the solvent accessible active site entrance. However, Leu 38 and Lys 109 constrain the orientation of the coumaroyl moiety in the binding cleft. Architecturally, the CHI active site limits the substrate's available conformations to ensure stereospecific product formation.
Subtle variations in substrate preference reflected in the Km values for chalcone versus 6′-deoxychalcone exist between CHIs of different species (Dixon et al. Phytochemistry 27:2801-2808, 1988). CHIs from legumes, such as alfalfa, prefer 6′-deoxychalcone as a substrate but the enzymes from non-legumes, like petunia, optimally use chalcone. The structure of the CHI•naringenin complex (SEQ ID NO:9), viewed with reference to the amino acid sequences of different CHIs, show that Thr 190 and Met 191 may partially modulate substrate preference. In the CHIs from non-legumes, a serine and an isoleucine replace Thr 190 and Met 191 (see SEQ ID NO:1), respectively. These two differences may better accommodate the 6′-hydroxyl moiety of chalcone due to a modest increase in active site volume in the vicinity of the trihydroxyl ring.
The present invention provides for the first time the intramolecular reaction of CHI (SEQ ID NOs:1, 9-11) with its product. CHI catalyzes an intramolecular reaction utilizing a substrate-derived nucleophile and a carbon-carbon double bond as a Michael acceptor. Two reaction mechanisms have been proposed for (2S)-naringenin formation by CHI. One involves nucleophilic catalysis by an active site residue that forms a covalent intermediate that is released after a SN2 displacement by the 2′-O− of chalcone. The other mechanism invokes general acid-base catalysis employing an enol intermediate. The structure of CHI clearly supports the latter mechanism.
Examination of the CHI•naringenin complex structure (SEQ ID NO:9) reveals a hydrogen bond network at the bottom of the binding cleft centered about the water molecule that contacts the ketone of (2S)-naringenin (FIGS. 4A and 4B). Of the five amino acids contributing to this network, only Thr 48 and Tyr 106 are conserved in all CHIs (SEQ ID NOs:1-8). The position of the water molecule between (2S)-naringenin and Tyr 106 (see SEQ ID NOs:1, 9-11) suggests a reaction mechanism in which the tyrosine activates the water, allowing it to serve as a general acid in the cyclization reaction (FIG. 4C). In the proposed reaction mechanism, the 2′-O− (pKa ̃7-8) forms in solution as suggested by studies on the spontaneous cyclization of chalcones. The negatively charged oxygen then attacks the carbon-carbon double bond of chalcone utilizing a Michael addition with the water molecule at the backside of the active site acting as the general acid in the transient protonation of the intermediate enolate.
Accordingly, for the first time, the invention provides the ability to modulate activity of the active site of CHI to design novel enzymes to catalyze the synthesis of various flavanones. For example, Tyr 106 (see SEQ ID NO:1) was substituted by phenylalanine and the properties of the mutant CHI (SEQ ID NO:9) compared to the wild-type enzyme. The present invention allows the comparison of the activities of mutants and designed mutants by computer modeling as well as by biological assays. The kinetics for the cyclization of 6′-deoxychalcone by wild-type CHI (SEQ ID NO:1) (kcat=4384 min−1; Km=25.7 μM; kcat/Km=1.71×108 M−1 min−1) versus those of the reaction catalyzed by the CHI Y106F mutant (see SEQ ID NO:1) (kcat=69.0 min−1; Km=29.1 μM; kcat/Km=2.37×106 M−1 min−1) demonstrate that the tyrosine residue contributes to the stabilization of the transition state. The 100-fold reduction in reaction rate is consistent with the decrease in rate associated with the loss of a general acid. However, the observed reaction rate with the mutant remains greater than that of the uncatalyzed cyclization reaction. Thus, the present invention demonstrates that the structural complementarity of the binding cleft to the transition state of the reaction contributes additional levels of catalytic rate enhancement.
A major contribution to rate enhancement in enzymatic reactions results from bringing substrates or reactive centers in the same molecule together in space. As described above, the topology of the binding cleft limits the flexibility of chalcone and eliminates catalytically unproductive orientations by spatially defining an optimal geometry for (2S)-naringenin formation. This effectively channels the ground state conformation of the substrate into a catalytically productive conformation. Together with contributions from general acid-base catalysis, shape complementarity between the CHI binding pocket and chalcone accelerates the cyclization of chalcone 107-fold over the spontaneous reaction rate. Accordingly, the present invention, provides for the first time, the ability to design, model, and assay native CHI and mutant CHI polypeptides.
The three-dimensional structure of CHI (see SEQ ID NOs:1, 9-11), provided herein, together with the structure of chalcone synthase (see WO/01/07579 A2, published Feb. 1, 2001), provides a useful template for engineering experiments that aim to diversify and modify flavonoid biosynthetic pathways for crop and food sources, as well as providing novel flavanones for intermediates and leads in drug discovery.
As used herein, “naturally occurring amino acid” and “naturally occurring R-group” includes L-isomers of the twenty amino acids naturally occurring in proteins. 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, arginine, and lysine. Unless specially indicated, all amino acids referred to in this application are in the L-form.
“Unnatural amino acid” and “unnatural R-group” includes amino acids that are not naturally found in proteins. Examples of unnatural amino acids included herein are racemic mixtures of selenocysteine and selenomethionine. In addition, unnatural amino acids include the D or L forms of, for example, nor-leucine, para-nitrophenylalanine, homophenylalanine, para-fluorophenylalanine, 3-amino-2-benzylpropionic acid, homoarginines, D-phenylalanine, and the like.
“R-group” refers to the substituent attached to the α-carbon of an amino acid residue. An R-group is an important determinant of the overall chemical character of an amino acid. There are twenty natural R-groups found in proteins, which make up the twenty naturally occurring amino acids.
“α-carbon” refers to the chiral carbon atom found in an amino acid residue. Typically, four substituents will be covalently bound to said α-carbon including an amine group, a carboxylic acid group, a hydrogen atom, and an R-group.
“Positively charged amino acid” and “positively charged R-group” includes any naturally occurring or unnatural amino acid having a positively charged side chain under normal physiological conditions. Examples of positively charged, naturally occurring amino acids include arginine, lysine, histidine, and the like.
“Negatively charged amino acid” and “negatively charged R-group” includes any naturally occurring or unnatural amino acid having a negatively charged side chain under normal physiological conditions. Examples of negatively charged, naturally occurring amino acids include aspartic acid, glutamic acid, and the like.
“Hydrophobic amino acid” and “hydrophobic R-group” includes any naturally occurring or unnatural amino acid 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, methionine, and the like.
“Hydrophilic amino acid” and “hydrophilic R-group” includes any naturally occurring or unnatural amino acid having a charged polar side chain that is relatively soluble in water. Examples of naturally occurring hydrophilic amino acids include serine, threonine, tyrosine, asparagine, glutamine, cysteine, and the like.
“Mutant” or “mutated isomerase” refers to an isomerase enzyme (e.g., chalcone isomerase) having one or more R-group modifications to the amino acids of a wild-type isomerase or having a substitution of one or more amino acids, either conservative or non-conservative substitutions, that result in a modification to the catalytic activity of a wild-type isomerase. For example, a mutant isomerase has an R-group on one or more α-carbon other than the prescribed arrangements of R-groups associated with one or more α-carbon of a known isolated chalcone isomerase (Accession No. 1EYP, Protein Data Bank, Table 1, SEQ ID NOs:9-11). Access to the foregoing information in the Protein Data Bank can be found on the World Wide Web at the website for rcsb.org. Typically mutants refer to changes or modification to the configuration of R-groups within the active site, however mutations outside of the residues found in the active site are also considered to be mutants in light of the present invention.
“Nonmutated isomerase” includes an isomerase wherein no R-group(s) are changed relative to the active site of CHI (see, for example, PDB Accession No. 1EYP; and Table 1, SEQ ID NOs:9-11). A nonmutated isomerase according to the present invention may or may not have amino acid residues outside of the active site that are the same as those taught for native CHI (SEQ ID NO:1).
The R-groups of known isolated chalcone isomerases can be readily determined by consulting sequence databases well known in the art such as, for example, GenBank. Additional R-groups found inside and/or outside of the active site may or may not be the same. R-groups may be a natural R-group, unnatural R-group, hydrophobic R-group, hydrophilic R-group, positively charged R-group, negatively charged R-group, and the like.
“Non-native” or “non-native isomerase” refers to an isomerase protein that is not found in nature, whether isolated or not. A non-native isomerase may, for example, be a mutated isomerase (see, the Examples below).
“Native” or “native isomerase” refers to isomerase proteins that are produced in nature, e.g., are not mutated (see, for example, PDB Accession No. 1EYP, SEQ ID NOs:9-11).
“Purified” or “isolated” refers to a protein or nucleic acid, respectively, that has been separated from its natural environment Contaminant components of its natural environment may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes. In one embodiment, the isolated molecule, in the case of a protein, will be purified to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence or to homogeneity by SDS-PAGE under reducing or non-reducing conditions using Coomassie blue or silver stain. In the case of a nucleic acid the isolated molecule will preferably be purified to a degree sufficient to obtain a nucleic acid sequence using standard sequencing methods.
By a “substantially pure polypeptide” is meant an isomerase polypeptide (e.g., a chalcone isomerase) which has been separated from components which naturally accompany it. Typically, the polypeptide is substantially pure when it is at least 60%, by weight, free from the proteins and naturally-occurring organic molecules with which it is naturally associated. Preferably, the preparation is at least 75%, more preferably at least 90%, and most preferably at least 99%, by weight, isomerase polypeptide. A substantially pure isomerase polypeptide may be obtained, for example, by extraction from a natural source; by expression of a recombinant nucleic acid encoding an isomerase polypeptide; or by chemically synthesizing the protein. Purity can be measured by any appropriate method (e.g., column chromatography, polyacrylamide gel electrophoresis, or by HPLC analysis).
“Degenerate variations thereof” refers to changing a gene sequence using the degenerate nature of the genetic code to encode proteins having the same amino acid sequence yet having a different gene sequence. For example, a chalcone isomerase of the present invention is based on amino acid sequences. Degenerate gene variations thereof can be made encoding the same protein due to the plasticity of the genetic code, as described herein.
“Expression” refers to transcription of a gene or nucleic acid sequence, stable accumulation of nucleic acid, and the translation of that nucleic acid to a polypeptide sequence. Expression of genes also involves transcription of the gene to make RNA, processing of RNA into mRNA in eukaryotic systems, and translation of mRNA into proteins. It is not necessary for the genes to integrate into the genome of a cell in order to achieve expression. This definition in no way limits expression to a particular system or to being confined to cells or a particular cell type and is meant to include cellular, transient, in vitro, in vivo, and viral expression systems in both prokaryotic, eukaryotic cells, and the like.
“Foreign” or “heterologous” genes refers to a gene encoding a protein whose exact amino acid sequence is not normally found in the host cell.
“Promoter” and “promoter regulatory element”, and the like, refers to a nucleotide sequence element within a nucleic acid fragment or gene that controls the expression of that gene. These can also include expression control sequences. Promoter regulatory elements, and the like, from a variety of sources can be used efficiently to promote gene expression. Promoter regulatory elements are meant to include constitutive, tissue-specific, developmental-specific, inducible, subgenomic promoters, and the like. Promoter regulatory elements may also include certain enhancer elements or silencing elements that improve or regulate transcriptional efficiency. Promoter regulatory elements are recognized by RNA polymerases, promote the binding thereof, and facilitate RNA transcription.
Table 1 lists the atomic structure coordinates for a chalcone isomerase (SEQ ID NOs:9-11) as derived by X-ray diffraction from a crystal of a chalcone isomerase. The data set, which may also be referred to as the “atomic coordinates” or “structure coordinates”, is useful for the methods of the present invention. In addition, invention methods may use a subset or portion of the atomic coordinates contained within this data set, for example, those atomic coordinates defining the amino acid residues which comprise the enzymatic active site. The following abbreviations are used in Table 1: “Atom Type” refers to the element whose coordinates are measured. The first letter in the column defines the element; “X, Y, Z” crystallographically define the atomic position of the element measured; “B” is a thermal factor that measures movement of the atom around its atomic center; and “molecule” denoted in the table refers to the particular monomer of CHI.
















TABLE 1






Atom










Atom
Type
Res
#
X
Y
Z
OCC
B
Mol.
























1
CB
SER
4
1.314
23.727
49.727
1.00
67.92
A


2
OG
SER
4
1.884
22.898
50.729
1.00
68.17
A


3
C
SER
4
2.569
22.924
47.696
1.00
67.57
A


4
O
SER
4
3.457
23.749
47.948
1.00
67.60
A


5
N
SER
4
.637
21.623
48.600
1.00
67.80
A


6
CA
SER
4
1.205
22.987
48.386
1.00
67.75
A


7
N
ILE
5
2.718
21.924
46.829
1.00
67.22
A


8
CA
ILE
5
3.933
21.713
46.044
1.00
66.80
A


9
CB
ILE
5
4.100
20.197
45.707
1.00
66.89
A


10
CG2
ILE
5
2.735
19.531
45.616
1.00
66.97
A


11
CG1
ILE
5
4.885
20.005
44.408
1.00
66.89
A


12
CD1
ILE
5
6.348
20.312
44.527
1.00
66.95
A


13
C
ILE
5
3.731
22.543
44.776
1.00
66.40
A


14
O
ILE
5
2.987
22.154
43.877
1.00
66.38
A


15
N
THR
6
4.382
23.700
44.723
1.00
65.89
A


16
CA
THR
6
4.247
24.615
43.594
1.00
65.37
A


17
CB
THR
6
4.518
26.068
44.030
1.00
65.36
A


18
OG1
THR
6
5.842
26.163
44.573
1.00
65.19
A


19
CG2
THR
6
3.510
26.509
45.078
1.00
65.29
A


20
C
THR
6
5.149
24.331
42.402
1.00
65.02
A


21
O
THR
6
5.990
23.432
42.429
1.00
64.94
A


22
N
ALA
7
4.947
25.113
41.347
1.00
64.56
A


23
CA
ALA
7
5.747
25.001
40.141
1.00
64.14
A


24
CB
ALA
7
4.900
25.305
38.911
1.00
64.10
A


25
C
ALA
7
6.848
26.040
40.294
1.00
63.80
A


26
O
ALA
7
6.735
26.954
41.114
1.00
63.75
A


27
N
ILE
8
7.917
25.894
39.524
1.00
63.43
A


28
CA
ILE
8
9.020
26.841
39.589
1.00
63.05
A


29
CB
ILE
8
10.269
26.220
40.250
1.00
62.98
A


30
CG2
ILE
8
11.387
27.252
40.320
1.00
62.92
A


31
CG1
ILE
8
9.926
25.724
41.654
1.00
62.91
A


32
CD1
ILE
8
11.064
25.002
42.332
1.00
62.85
A


33
C
ILE
8
9.397
27.283
38.186
1.00
62.84
A


34
O
ILE
8
9.495
26.467
37.272
1.00
62.74
A


35
N
THR
9
9.598
28.581
38.015
1.00
62.62
A


36
CA
THR
9
9.984
29.104
36.716
1.00
62.45
A


37
CB
THR
9
9.017
30.206
36.235
1.00
62.45
A


38
OG1
THR
9
7.720
29.637
36.021
1.00
62.47
A


39
CG2
THR
9
9.505
30.817
34.928
1.00
62.44
A


40
C
THR
9
11.393
29.664
36.802
1.00
62.27
A


41
O
THR
9
11.708
30.455
37.689
1.00
62.24
A


42
N
VAL
10
12.244
29.225
35.885
1.00
62.10
A


43
CA
VAL
10
13.624
29.681
35.835
1.00
61.94
A


44
CB
VAL
10
14.608
28.509
36.024
1.00
61.87
A


45
CG1
VAL
10
16.039
29.011
35.927
1.00
61.79
A


46
CG2
VAL
10
14.369
27.839
37.370
1.00
61.80
A


47
C
VAL
10
13.863
30.310
34.469
1.00
61.89
A


48
O
VAL
10
13.782
29.627
33.448
1.00
61.89
A


49
N
GLU
11
14.148
31.608
34.453
1.00
61.82
A


50
CA
GLU
11
14.390
32.314
33.201
1.00
61.74
A


51
CB
GLU
11
15.757
31.931
32.634
1.00
61.91
A


52
CG
GLU
11
16.926
32.601
33.322
1.00
62.24
A


53
CD
GLU
11
16.947
34.102
33.095
1.00
62.41
A


54
OE1
GLU
11
16.966
34.530
31.919
1.00
62.59
A


55
OE2
GLU
11
16.944
34.851
34.093
1.00
62.48
A


56
C
GLU
11
13.310
32.001
32.173
1.00
61.59
A


57
O
GLU
11
13.609
31.632
31.039
1.00
61.66
A


58
N
ASN
12
12.054
32.132
32.583
1.00
61.41
A


59
CA
ASN
12
10.922
31.878
31.698
1.00
61.23
A


60
CB
ASN
12
11.060
32.716
30.426
1.00
61.51
A


61
CG
ASN
12
11.305
34.181
30.726
1.00
61.77
A


62
OD1
ASN
12
10.511
34.824
31.417
1.00
61.95
A


63
ND2
ASN
12
12.415
34.718
30.214
1.00
61.87
A


64
C
ASN
12
10.728
30.408
31.330
1.00
60.92
A


65
O
ASN
12
9.832
30.075
30.559
1.00
60.93
A


66
N
LEU
13
11.576
29.536
31.865
1.00
60.46
A


67
CA
LEU
13
11.451
28.103
31.612
1.00
60.03
A


68
CB
LEU
13
12.832
27.448
31.513
1.00
59.92
A


69
CG
LEU
13
13.692
27.886
30.326
1.00
59.82
A


70
CD1
LEU
13
15.112
27.380
30.496
1.00
59.84
A


71
CD2
LEU
13
13.082
27.359
29.041
1.00
59.83
A


72
C
LEU
13
10.690
27.559
32.813
1.00
59.78
A


73
O
LEU
13
11.206
27.551
33.930
1.00
59.74
A


74
N
GLU
14
9.460
27.115
32.595
1.00
59.52
A


75
CA
GLU
14
8.664
26.616
33.705
1.00
59.29
A


76
CB
GLU
14
7.178
26.915
33.491
1.00
59.60
A


77
CG
GLU
14
6.319
26.482
34.673
1.00
60.12
A


78
CD
GLU
14
4.845
26.394
34.340
1.00
60.47
A


79
OE1
GLU
14
4.490
25.643
33.400
1.00
60.76
A


80
OE2
GLU
14
4.041
27.067
35.023
1.00
60.61
A


81
C
GLU
14
8.818
25.133
33.989
1.00
58.89
A


82
O
GLU
14
8.951
24.315
33.078
1.00
58.84
A


83
N
TYR
15
8.796
24.811
35.276
1.00
58.40
A


84
CA
TYR
15
8.894
23.442
35.750
1.00
58.01
A


85
CB
TYR
15
10.107
23.270
36.663
1.00
57.57
A


86
CG
TYR
15
11.404
23.193
35.903
1.00
57.20
A


87
CD1
TYR
15
11.998
24.336
35.373
1.00
56.96
A


88
CE1
TYR
15
13.172
24.256
34.631
1.00
56.83
A


89
CD2
TYR
15
12.017
21.964
35.674
1.00
56.98
A


90
CE2
TYR
15
13.186
21.870
34.934
1.00
56.81
A


91
CZ
TYR
15
13.760
23.016
34.414
1.00
56.76
A


92
OH
TYR
15
14.919
22.915
33.679
1.00
56.45
A


93
C
TYR
15
7.618
23.132
36.516
1.00
57.93
A


94
O
TYR
15
7.439
23.577
37.647
1.00
57.80
A


95
N
PRO
16
6.703
22.379
35.891
1.00
57.97
A


96
CD
PRO
16
6.811
21.806
34.539
1.00
57.95
A


97
CA
PRO
16
5.429
22.005
36.514
1.00
58.06
A


98
CB
PRO
16
4.829
21.029
35.505
1.00
58.00
A


99
CG
PRO
16
5.373
21.514
34.206
1.00
58.02
A


100
C
PRO
16
5.666
21.351
37.872
1.00
58.14
A


101
O
PRO
16
6.686
20.694
38.081
1.00
58.11
A


102
N
ALA
17
4.722
21.532
38.789
1.00
58.25
A


103
CA
ALA
17
4.839
20.957
40.123
1.00
58.39
A


104
CB
ALA
17
3.678
21.419
40.990
1.00
58.35
A


105
C
ALA
17
4.872
19.435
40.067
1.00
58.46
A


106
O
ALA
17
5.458
18.786
40.932
1.00
58.51
A


107
N
VAL
18
4.242
18.872
39.045
1.00
58.58
A


108
CA
VAL
18
4.183
17.425
38.882
1.00
58.75
A


109
CB
VAL
18
2.849
16.865
39.448
1.00
58.76
A


110
CG1
VAL
18
2.686
15.402
39.079
1.00
58.85
A


111
CG2
VAL
18
2.821
17.025
40.957
1.00
58.73
A


112
C
VAL
18
4.296
17.049
37.412
1.00
58.83
A


113
O
VAL
18
3.944
17.835
36.535
1.00
58.87
A


114
N
VAL
19
4.800
15.850
37.147
1.00
58.92
A


115
CA
VAL
19
4.942
15.359
35.782
1.00
59.03
A


116
CB
VAL
19
6.300
15.763
35.148
1.00
59.07
A


117
CG1
VAL
19
6.430
17.275
35.089
1.00
59.10
A


118
CG2
VAL
19
7.445
15.160
35.934
1.00
59.07
A


119
C
VAL
19
4.863
13.841
35.794
1.00
59.13
A


120
O
VAL
19
5.164
13.207
36.808
1.00
59.11
A


121
N
THR
20
4.456
13.266
34.666
1.00
59.26
A


122
CA
THR
20
4.350
11.817
34.529
1.00
59.38
A


123
CB
THR
20
2.899
11.393
34.226
1.00
59.40
A


124
OG1
THR
20
2.067
11.731
35.342
1.00
59.47
A


125
CG2
THR
20
2.819
9.890
33.973
1.00
59.42
A


126
C
THR
20
5.258
11.353
33.395
1.00
59.42
A


127
O
THR
20
5.264
11.943
32.317
1.00
59.50
A


128
N
SER
21
6.032
10.301
33.640
1.00
59.49
A


129
CA
SER
21
6.943
9.784
32.625
1.00
59.51
A


130
CB
SER
21
8.044
8.942
33.271
1.00
59.56
A


131
OG
SER
21
8.853
8.329
32.277
1.00
59.58
A


132
C
SER
21
6.247
8.942
31.566
1.00
59.51
A


133
O
SER
21
5.539
7.989
31.884
1.00
59.49
A


134
N
PRO
22
6.427
9.299
30.287
1.00
59.53
A


135
CD
PRO
22
6.896
10.609
29.803
1.00
59.57
A


136
CA
PRO
22
5.805
8.540
29.199
1.00
59.44
A


137
CB
PRO
22
5.976
9.463
27.991
1.00
59.52
A


138
CG
PRO
22
6.003
10.835
28.609
1.00
59.57
A


139
C
PRO
22
6.576
7.232
29.029
1.00
59.35
A


140
O
PRO
22
6.142
6.322
28.321
1.00
59.29
A


141
N
VAL
23
7.722
7.152
29.701
1.00
59.24
A


142
CA
VAL
23
8.586
5.980
29.632
1.00
59.12
A


143
CB
VAL
23
10.076
6.383
29.781
1.00
59.16
A


144
CG1
VAL
23
10.967
5.150
29.690
1.00
59.11
A


145
CG2
VAL
23
10.450
7.400
28.709
1.00
59.19
A


146
C
VAL
23
8.272
4.924
30.689
1.00
59.03
A


147
O
VAL
23
8.253
3.731
30.393
1.00
59.06
A


148
N
THR
24
8.019
5.362
31.918
1.00
58.90
A


149
CA
THR
24
7.748
4.432
33.008
1.00
58.69
A


150
CB
THR
24
8.759
4.636
34.150
1.00
58.76
A


151
OG1
THR
24
8.694
5.995
34.596
1.00
58.70
A


152
CG2
THR
24
10.174
4.327
33.678
1.00
58.73
A


153
C
THR
24
6.345
4.544
33.595
1.00
58.51
A


154
O
THR
24
5.917
3.675
34.356
1.00
58.54
A


155
N
GLY
25
5.636
5.612
33.252
1.00
58.25
A


156
CA
GLY
25
4.293
5.791
33.775
1.00
57.90
A


157
C
GLY
25
4.290
6.303
35.205
1.00
57.62
A


158
O
GLY
25
3.239
6.623
35.764
1.00
57.68
A


159
N
LYS
26
5.472
6.383
35.805
1.00
57.25
A


160
CA
LYS
26
5.592
6.862
37.174
1.00
56.78
A


161
CB
LYS
26
6.977
6.513
37.720
1.00
56.84
A


162
CG
LYS
26
7.244
5.014
37.718
1.00
56.94
A


163
CD
LYS
26
8.619
4.652
38.255
1.00
57.05
A


164
CE
LYS
26
8.748
3.143
38.402
1.00
57.13
A


165
NZ
LYS
26
10.088
2.707
38.901
1.00
57.35
A


166
C
LYS
26
5.354
8.368
37.224
1.00
56.40
A


167
O
LYS
26
5.589
9.078
36.243
1.00
56.47
A


168
N
SER
27
4.860
8.853
38.357
1.00
55.84
A


169
CA
SER
27
4.607
10.282
38.512
1.00
55.24
A


170
CB
SER
27
3.182
10.515
39.021
1.00
55.40
A


171
OG
SER
27
2.947
9.774
40.205
1.00
55.82
A


172
C
SER
27
5.619
10.866
39.491
1.00
54.61
A


173
O
SER
27
6.023
10.197
40.443
1.00
54.55
A


174
N
TYR
28
6.032
12.108
39.252
1.00
53.92
A


175
CA
TYR
28
7.010
12.759
40.120
1.00
53.08
A


176
CB
TYR
28
8.364
12.920
39.414
1.00
53.09
A


177
CG
TYR
28
8.740
11.829
38.440
1.00
53.03
A


178
CD1
TYR
28
8.118
11.735
37.199
1.00
53.08
A


179
CE1
TYR
28
8.493
10.762
36.282
1.00
53.18
A


180
CD2
TYR
28
9.748
10.915
38.744
1.00
53.03
A


181
CE2
TYR
28
10.129
9.938
37.837
1.00
52.99
A


182
CZ
TYR
28
9.500
9.867
36.608
1.00
53.14
A


183
OH
TYR
28
9.874
8.908
35.701
1.00
53.23
A


184
C
TYR
28
6.550
14.146
40.541
1.00
52.56
A


185
O
TYR
28
5.671
14.743
39.916
1.00
52.53
A


186
N
PHE
29
7.155
14.658
41.606
1.00
51.84
A


187
CA
PHE
29
6.837
15.992
42.082
1.00
51.21
A


188
CB
PHE
29
6.298
15.937
43.517
1.00
51.18
A


189
CG
PHE
29
7.297
15.469
44.535
1.00
51.12
A


190
CD1
PHE
29
8.117
16.379
45.195
1.00
51.10
A


191
CD2
PHE
29
7.404
14.119
44.852
1.00
51.11
A


192
CE1
PHE
29
9.030
15.954
46.160
1.00
51.04
A


193
CE2
PHE
29
8.315
13.683
45.816
1.00
51.11
A


194
CZ
PHE
29
9.129
14.604
46.471
1.00
51.04
A


195
C
PHE
29
8.126
16.801
42.004
1.00
50.74
A


196
O
PHE
29
9.218
16.260
42.184
1.00
50.60
A


197
N
LEU
30
8.003
18.087
41.710
1.00
50.27
A


198
CA
LEU
30
9.172
18.951
41.601
1.00
49.82
A


199
CB
LEU
30
8.757
20.321
41.069
1.00
49.72
A


200
CG
LEU
30
9.886
21.320
40.830
1.00
49.58
A


201
CD1
LEU
30
10.840
20.777
39.779
1.00
49.42
A


202
CD2
LEU
30
9.284
22.653
40.395
1.00
49.59
A


203
C
LEU
30
9.874
19.111
42.945
1.00
49.54
A


204
O
LEU
30
9.311
19.671
43.887
1.00
49.44
A


205
N
GLY
31
11.106
18.613
43.024
1.00
49.23
A


206
CA
GLY
31
11.866
18.724
44.257
1.00
48.92
A


207
C
GLY
31
12.561
20.067
44.333
1.00
48.73
A


208
O
GLY
31
12.777
20.617
45.414
1.00
48.72
A


209
N
GLY
32
12.904
20.601
43.167
1.00
48.61
A


210
CA
GLY
32
13.571
21.885
43.091
1.00
48.42
A


211
C
GLY
32
14.033
22.199
41.681
1.00
48.30
A


212
O
GLY
32
14.156
21.306
40.841
1.00
48.08
A


213
N
ALA
33
14.278
23.479
41.422
1.00
48.27
A


214
CA
ALA
33
14.738
23.938
40.118
1.00
48.30
A


215
CB
ALA
33
13.553
24.384
39.266
1.00
48.18
A


216
C
ALA
33
15.704
25.094
40.314
1.00
48.37
A


217
O
ALA
33
15.560
25.884
41.247
1.00
48.48
A


218
N
GLY
34
16.694
25.186
39.437
1.00
48.42
A


219
CA
GLY
34
17.667
26.252
39.529
1.00
48.50
A


220
C
GLY
34
18.320
26.486
38.181
1.00
48.60
A


221
O
GLY
34
17.731
26.209
37.132
1.00
48.47
A


222
N
GLU
35
19.550
26.984
38.208
1.00
48.66
A


223
CA
GLU
35
20.268
27.255
36.980
1.00
48.81
A


224
CB
GLU
35
20.173
28.731
36.625
1.00
49.27
A


225
CG
GLU
35
19.336
29.521
37.593
1.00
50.31
A


226
CD
GLU
35
19.328
30.995
37.273
1.00
50.76
A


227
OE1
GLU
35
19.983
31.383
36.294
1.00
51.26
A


228
OE2
GLU
35
18.668
31.766
37.995
1.00
51.30
A


229
C
GLU
35
21.724
26.865
37.061
1.00
48.56
A


230
O
GLU
35
22.319
26.835
38.137
1.00
48.49
A


231
N
ARG
36
22.294
26.557
35.906
1.00
48.31
A


232
CA
ARG
36
23.692
26.184
35.814
1.00
48.10
A


233
CB
ARG
36
23.834
24.681
35.559
1.00
48.15
A


234
CG
ARG
36
24.466
23.912
36.716
1.00
48.32
A


235
CD
ARG
36
23.936
24.429
38.027
1.00
48.33
A


236
NE
ARG
36
23.980
23.448
39.103
1.00
48.32
A


237
CZ
ARG
36
23.172
23.506
40.153
1.00
48.28
A


238
NH1
ARG
36
22.287
24.492
40.229
1.00
48.19
A


239
NH2
ARG
36
23.239
22.590
41.114
1.00
48.15
A


240
C
ARG
36
24.256
26.954
34.648
1.00
48.05
A


241
O
ARG
36
23.530
27.321
33.721
1.00
47.90
A


242
N
GLY
37
25.553
27.201
34.706
1.00
47.95
A


243
CA
GLY
37
26.202
27.934
33.646
1.00
47.99
A


244
C
GLY
37
27.680
27.972
33.926
1.00
47.99
A


245
O
GLY
37
28.225
27.045
34.519
1.00
47.95
A


246
N
LEU
38
28.328
29.052
33.510
1.00
47.97
A


247
CA
LEU
38
29.757
29.190
33.723
1.00
47.98
A


248
CB
LEU
38
30.497
29.063
32.382
1.00
48.09
A


249
CG
LEU
38
30.242
27.777
31.580
1.00
48.20
A


250
CD1
LEU
38
30.780
27.914
30.165
1.00
48.09
A


251
CD2
LEU
38
30.894
26.601
32.287
1.00
48.21
A


252
C
LEU
38
30.071
30.533
34.361
1.00
47.94
A


253
O
LEU
38
29.368
31.521
34.145
1.00
47.77
A


254
N
THR
39
31.114
30.554
35.178
1.00
47.97
A


255
CA
THR
39
31.544
31.794
35.797
1.00
48.05
A


256
CB
THR
39
32.021
31.579
37.242
1.00
48.08
A


257
OG1
THR
39
30.921
31.111
38.030
1.00
48.06
A


258
CG2
THR
39
32.532
32.894
37.839
1.00
48.01
A


259
C
THR
39
32.697
32.254
34.919
1.00
48.04
A


260
O
THR
39
33.765
31.642
34.902
1.00
48.00
A


261
N
ILE
40
32.452
33.313
34.160
1.00
48.10
A


262
CA
ILE
40
33.452
33.854
33.249
1.00
48.20
A


263
CB
ILE
40
32.884
33.983
31.816
1.00
48.29
A


264
CG2
ILE
40
33.992
34.369
30.856
1.00
48.42
A


265
CG1
ILE
40
32.241
32.664
31.372
1.00
48.43
A


266
CD1
ILE
40
33.229
31.523
31.159
1.00
48.46
A


267
C
ILE
40
33.858
35.241
33.732
1.00
48.15
A


268
O
ILE
40
33.030
36.156
33.785
1.00
48.07
A


269
N
GLU
41
35.131
35.389
34.085
1.00
48.08
A


270
CA
GLU
41
35.644
36.665
34.569
1.00
47.96
A


271
CB
GLU
41
35.594
37.711
33.450
1.00
48.36
A


272
CG
GLU
41
36.387
37.321
32.207
1.00
48.95
A


273
CD
GLU
41
37.889
37.463
32.393
1.00
49.41
A


274
OE1
GLU
41
38.413
36.986
33.424
1.00
49.70
A


275
OE2
GLU
41
38.549
38.051
31.502
1.00
49.86
A


276
C
GLU
41
34.816
37.137
35.754
1.00
47.62
A


277
O
GLU
41
34.394
38.290
35.804
1.00
47.61
A


278
N
GLY
42
34.575
36.234
36.700
1.00
47.21
A


279
CA
GLY
42
33.805
36.586
37.881
1.00
46.69
A


280
C
GLY
42
32.329
36.848
37.638
1.00
46.35
A


281
O
GLY
42
31.622
37.266
38.555
1.00
46.46
A


282
N
ASN
43
31.861
36.603
36.415
1.00
45.84
A


283
CA
ASN
43
30.458
36.809
36.057
1.00
45.29
A


284
CB
ASN
43
30.349
37.654
34.786
1.00
45.16
A


285
CG
ASN
43
30.617
39.121
35.035
1.00
45.00
A


286
OD1
ASN
43
29.693
39.899
35.273
1.00
44.96
A


287
ND2
ASN
43
31.886
39.506
34.990
1.00
44.74
A


288
C
ASN
43
29.760
35.479
35.813
1.00
45.05
A


289
O
ASN
43
30.271
34.628
35.083
1.00
44.95
A


290
N
PHE
44
28.592
35.296
36.421
1.00
44.77
A


291
CA
PHE
44
27.856
34.062
36.218
1.00
44.54
A


292
CB
PHE
44
26.857
33.801
37.345
1.00
44.68
A


293
CG
PHE
44
26.102
32.502
37.187
1.00
44.83
A


294
CD1
PHE
44
26.780
31.286
37.185
1.00
44.87
A


295
CD2
PHE
44
24.719
32.496
37.048
1.00
44.98
A


296
CE1
PHE
44
26.092
30.079
37.050
1.00
45.03
A


297
CE2
PHE
44
24.019
31.298
36.914
1.00
45.00
A


298
CZ
PHE
44
24.708
30.084
36.915
1.00
45.08
A


299
C
PHE
44
27.100
34.142
34.904
1.00
44.28
A


300
O
PHE
44
26.204
34.968
34.739
1.00
44.16
A


301
N
ILE
45
27.477
33.284
33.967
1.00
44.04
A


302
CA
ILE
45
26.825
33.245
32.670
1.00
43.98
A


303
CB
ILE
45
27.867
33.142
31.535
1.00
43.86
A


304
CG2
ILE
45
27.179
33.253
30.171
1.00
43.72
A


305
CG1
ILE
45
28.915
34.253
31.697
1.00
43.65
A


306
CD1
ILE
45
28.339
35.658
31.718
1.00
43.39
A


307
C
ILE
45
25.924
32.016
32.678
1.00
44.04
A


308
O
ILE
45
26.402
30.882
32.703
1.00
43.96
A


309
N
LYS
46
24.620
32.260
32.685
1.00
44.19
A


310
CA
LYS
46
23.632
31.191
32.713
1.00
44.41
A


311
CB
LYS
46
22.255
31.745
33.069
1.00
44.81
A


312
CG
LYS
46
22.226
32.729
34.226
1.00
45.69
A


313
CD
LYS
46
20.797
33.163
34.535
1.00
46.28
A


314
CE
LYS
46
20.057
33.662
33.294
1.00
46.83
A


315
NZ
LYS
46
20.028
35.167
33.162
1.00
47.55
A


316
C
LYS
46
23.540
30.499
31.359
1.00
44.31
A


317
O
LYS
46
23.489
31.161
30.326
1.00
44.22
A


318
N
PHE
47
23.517
29.169
31.376
1.00
44.11
A


319
CA
PHE
47
23.410
28.391
30.151
1.00
44.14
A


320
CB
PHE
47
24.660
27.550
29.933
1.00
43.96
A


321
CG
PHE
47
25.743
28.279
29.203
1.00
44.12
A


322
CD1
PHE
47
26.644
29.092
29.888
1.00
44.02
A


323
CD2
PHE
47
25.827
28.201
27.815
1.00
44.07
A


324
CE1
PHE
47
27.611
29.817
29.200
1.00
43.92
A


325
CE2
PHE
47
26.791
28.923
27.116
1.00
43.99
A


326
CZ
PHE
47
27.686
29.735
27.811
1.00
43.99
A


327
C
PHE
47
22.194
27.484
30.150
1.00
44.13
A


328
O
PHE
47
21.664
27.152
29.086
1.00
44.05
A


329
N
THR
48
21.743
27.077
31.330
1.00
44.21
A


330
CA
THR
48
20.590
26.200
31.393
1.00
44.44
A


331
CB
THR
48
21.002
24.726
31.133
1.00
44.49
A


332
OG1
THR
48
19.824
23.910
31.060
1.00
44.77
A


333
CG2
THR
48
21.902
24.200
32.253
1.00
44.35
A


334
C
THR
48
19.835
26.247
32.702
1.00
44.49
A


335
O
THR
48
20.341
26.708
33.728
1.00
44.53
A


336
N
ALA
49
18.596
25.789
32.643
1.00
44.62
A


337
CA
ALA
49
17.757
25.700
33.819
1.00
44.66
A


338
CB
ALA
49
16.326
26.106
33.502
1.00
44.62
A


339
C
ALA
49
17.821
24.218
34.149
1.00
44.73
A


340
O
ALA
49
18.098
23.387
33.277
1.00
44.64
A


341
N
ILE
50
17.605
23.888
35.409
1.00
44.80
A


342
CA
ILE
50
17.630
22.501
35.823
1.00
45.04
A


343
CB
ILE
50
18.963
22.098
36.472
1.00
45.43
A


344
CG2
ILE
50
18.916
20.618
36.842
1.00
45.46
A


345
CG1
ILE
50
20.117
22.339
35.504
1.00
45.85
A


346
CD1
ILE
50
21.468
22.096
36.128
1.00
46.32
A


347
C
ILE
50
16.547
22.281
36.849
1.00
44.88
A


348
O
ILE
50
16.352
23.097
37.749
1.00
44.85
A


349
N
GLY
51
15.841
21.174
36.700
1.00
44.82
A


350
CA
GLY
51
14.792
20.836
37.637
1.00
44.61
A


351
C
GLY
51
14.951
19.380
38.020
1.00
44.59
A


352
O
GLY
51
15.251
18.535
37.175
1.00
44.53
A


353
N
VAL
52
14.772
19.086
39.301
1.00
44.48
A


354
CA
VAL
52
14.881
17.721
39.780
1.00
44.45
A


355
CB
VAL
52
15.888
17.607
40.936
1.00
44.45
A


356
CG1
VAL
52
15.957
16.162
41.424
1.00
44.49
A


357
CG2
VAL
52
17.257
18.077
40.475
1.00
44.55
A


358
C
VAL
52
13.527
17.233
40.267
1.00
44.39
A


359
O
VAL
52
12.932
17.822
41.170
1.00
44.42
A


360
N
TYR
53
13.035
16.267
39.649
1.00
44.36
A


361
CA
TYR
53
11.764
15.585
40.053
1.00
44.38
A


362
CB
TYR
53
10.873
15.298
38.841
1.00
44.42
A


363
CG
TYR
53
10.307
16.543
38.209
1.00
44.64
A


364
CD1
TYR
53
11.004
17.218
37.211
1.00
44.66
A


365
CE1
TYR
53
10.510
18.392
36.656
1.00
44.85
A


366
CD2
TYR
53
9.091
17.075
38.642
1.00
44.79
A


367
CE2
TYR
53
8.585
18.256
38.093
1.00
44.84
A


368
CZ
TYR
53
9.300
18.906
37.102
1.00
44.90
A


369
OH
TYR
53
8.814
20.070
36.550
1.00
44.98
A


370
C
TYR
53
12.019
14.294
40.809
1.00
44.31
A


371
O
TYR
53
12.924
13.535
40.473
1.00
44.16
A


372
N
LEU
54
11.222
14.057
41.842
1.00
44.38
A


373
CA
LEU
54
11.360
12.846
42.630
1.00
44.58
A


374
CB
LEU
54
11.602
13.196
44.098
1.00
44.57
A


375
CG
LEU
54
12.821
14.066
44.413
1.00
44.65
A


376
CD1
LEU
54
12.956
14.179
45.920
1.00
44.61
A


377
CD2
LEU
54
14.081
13.457
43.814
1.00
44.54
A


378
C
LEU
54
10.067
12.055
42.488
1.00
44.72
A


379
O
LEU
54
8.980
12.634
42.499
1.00
44.65
A


380
N
GLU
55
10.182
10.740
42.340
1.00
44.92
A


381
CA
GLU
55
8.999
9.905
42.202
1.00
45.23
A


382
CB
GLU
55
9.388
8.429
42.108
1.00
45.30
A


383
CG
GLU
55
8.289
7.553
41.512
1.00
45.53
A


384
CD
GLU
55
8.682
6.094
41.412
1.00
45.70
A


385
OE1
GLU
55
9.897
5.791
41.434
1.00
45.76
A


386
OE2
GLU
55
7.773
5.245
41.297
1.00
45.91
A


387
C
GLU
55
8.118
10.141
43.422
1.00
45.38
A


388
O
GLU
55
8.619
10.341
44.528
1.00
45.30
A


389
N
ASP
56
6.805
10.133
43.221
1.00
45.67
A


390
CA
ASP
56
5.875
10.365
44.320
1.00
45.96
A


391
CB
ASP
56
4.433
10.122
43.847
1.00
46.66
A


392
CG
ASP
56
4.256
8.790
43.135
1.00
47.25
A


393
OD1
ASP
56
3.175
8.582
42.539
1.00
47.89
A


394
OD2
ASP
56
5.180
7.946
43.162
1.00
47.80
A


395
C
ASP
56
6.178
9.531
45.565
1.00
45.79
A


396
O
ASP
56
6.049
10.014
46.690
1.00
45.83
A


397
N
ILE
57
6.605
8.290
45.371
1.00
45.59
A


398
CA
ILE
57
6.904
7.422
46.505
1.00
45.50
A


399
CB
ILE
57
7.136
5.975
46.039
1.00
45.50
A


400
CG2
ILE
57
5.880
5.456
45.326
1.00
45.59
A


401
CG1
ILE
57
8.342
5.915
45.097
1.00
45.25
A


402
CD1
ILE
57
8.695
4.520
44.636
1.00
45.00
A


403
C
ILE
57
8.107
7.875
47.339
1.00
45.50
A


404
O
ILE
57
8.345
7.355
48.428
1.00
45.45
A


405
N
ALA
58
8.861
8.847
46.837
1.00
45.47
A


406
CA
ALA
58
10.025
9.338
47.569
1.00
45.46
A


407
CB
ALA
58
10.709
10.441
46.777
1.00
45.36
A


408
C
ALA
58
9.636
9.852
48.956
1.00
45.54
A


409
O
ALA
58
10.403
9.735
49.915
1.00
45.38
A


410
N
VAL
59
8.433
10.414
49.053
1.00
45.66
A


411
CA
VAL
59
7.933
10.963
50.310
1.00
45.90
A


412
CB
VAL
59
6.510
11.543
50.125
1.00
45.96
A


413
CG1
VAL
59
6.036
12.200
51.414
1.00
46.02
A


414
CG2
VAL
59
6.513
12.547
48.981
1.00
46.03
A


415
C
VAL
59
7.911
9.917
51.418
1.00
45.95
A


416
O
VAL
59
8.474
10.132
52.493
1.00
46.04
A


417
N
ALA
60
7.259
8.789
51.156
1.00
46.03
A


418
CA
ALA
60
7.176
7.706
52.132
1.00
46.13
A


419
CB
ALA
60
6.321
6.570
51.576
1.00
46.10
A


420
C
ALA
60
8.567
7.181
52.487
1.00
46.26
A


421
O
ALA
60
8.842
6.844
53.642
1.00
46.27
A


422
N
SER
61
9.441
7.105
51.487
1.00
46.37
A


423
CA
SER
61
10.800
6.613
51.695
1.00
46.45
A


424
CB
SER
61
11.505
6.442
50.348
1.00
46.27
A


425
OG
SER
61
12.858
6.065
50.529
1.00
45.97
A


426
C
SER
61
11.634
7.534
52.584
1.00
46.70
A


427
O
SER
61
12.381
7.074
53.445
1.00
46.61
A


428
N
LEU
62
11.504
8.838
52.366
1.00
47.06
A


429
CA
LEU
62
12.264
9.824
53.124
1.00
47.50
A


430
CB
LEU
62
12.423
11.094
52.281
1.00
47.29
A


431
CG
LEU
62
13.216
10.963
50.974
1.00
47.14
A


432
CD1
LEU
62
12.898
12.134
50.065
1.00
47.11
A


433
CD2
LEU
62
14.707
10.901
51.269
1.00
47.05
A


434
C
LEU
62
11.645
10.181
54.475
1.00
47.94
A


435
O
LEU
62
12.342
10.636
55.382
1.00
47.96
A


436
N
ALA
63
10.341
9.959
54.606
1.00
48.47
A


437
CA
ALA
63
9.609
10.281
55.829
1.00
49.01
A


438
CB
ALA
63
8.182
9.738
55.735
1.00
48.91
A


439
C
ALA
63
10.248
9.812
57.134
1.00
49.39
A


440
O
ALA
63
10.470
10.619
58.043
1.00
49.51
A


441
N
ALA
64
10.545
8.523
57.227
1.00
49.79
A


442
CA
ALA
64
11.123
7.954
58.441
1.00
50.27
A


443
CB
ALA
64
11.583
6.537
58.170
1.00
50.33
A


444
C
ALA
64
12.273
8.767
59.020
1.00
50.57
A


445
O
ALA
64
12.213
9.230
60.164
1.00
50.74
A


446
N
LYS
65
13.314
8.954
58.226
1.00
50.78
A


447
CA
LYS
65
14.484
9.686
58.684
1.00
51.02
A


448
CB
LYS
65
15.705
9.308
57.832
1.00
50.84
A


449
CG
LYS
65
16.145
7.871
57.945
1.00
50.72
A


450
CD
LYS
65
17.253
7.556
56.959
1.00
50.55
A


451
CE
LYS
65
18.469
8.455
57.174
1.00
50.54
A


452
NZ
LYS
65
19.582
8.084
56.260
1.00
50.34
A


453
C
LYS
65
14.371
11.204
58.691
1.00
51.29
A


454
O
LYS
65
15.004
11.858
59.510
1.00
51.21
A


455
N
TRP
66
13.559
11.768
57.803
1.00
51.71
A


456
CA
TRP
66
13.510
13.224
57.702
1.00
52.27
A


457
CB
TRP
66
13.947
13.625
56.284
1.00
52.00
A


458
CG
TRP
66
15.219
12.925
55.918
1.00
51.86
A


459
CD2
TRP
66
16.500
13.127
56.532
1.00
51.78
A


460
CE2
TRP
66
17.380
12.174
55.983
1.00
51.69
A


461
CE3
TRP
66
16.983
14.018
57.494
1.00
51.76
A


462
CD1
TRP
66
15.376
11.897
55.039
1.00
51.80
A


463
NE1
TRP
66
16.676
11.435
55.075
1.00
51.70
A


464
CZ2
TRP
66
18.720
12.086
56.368
1.00
51.68
A


465
CZ3
TRP
66
18.316
13.928
57.878
1.00
51.76
A


466
CH2
TRP
66
19.166
12.964
57.310
1.00
51.70
A


467
C
TRP
66
12.265
14.000
58.099
1.00
52.82
A


468
O
TRP
66
12.299
15.230
58.109
1.00
52.83
A


469
N
LYS
67
11.172
13.321
58.418
1.00
53.49
A


470
CA
LYS
67
9.981
14.049
58.824
1.00
54.21
A


471
CB
LYS
67
8.819
13.085
59.067
1.00
54.43
A


472
CG
LYS
67
7.514
13.802
59.389
1.00
54.89
A


473
CD
LYS
67
6.372
12.831
59.655
1.00
55.21
A


474
CE
LYS
67
5.123
13.565
60.154
1.00
55.51
A


475
NZ
LYS
67
4.004
12.622
60.451
1.00
55.72
A


476
C
LYS
67
10.302
14.817
60.107
1.00
54.57
A


477
O
LYS
67
11.158
14.397
60.888
1.00
54.62
A


478
N
GLY
68
9.633
15.945
60.313
1.00
54.97
A


479
CA
GLY
68
9.881
16.714
61.517
1.00
55.48
A


480
C
GLY
68
11.142
17.567
61.493
1.00
55.79
A


481
O
GLY
68
11.321
18.427
62.351
1.00
55.94
A


482
N
LYS
69
12.024
17.344
60.528
1.00
56.06
A


483
CA
LYS
69
13.254
18.128
60.433
1.00
56.28
A


484
CB
LYS
69
14.303
17.396
59.586
1.00
56.40
A


485
CG
LYS
69
14.711
16.012
60.072
1.00
56.61
A


486
CD
LYS
69
15.423
16.035
61.404
1.00
56.82
A


487
CE
LYS
69
16.014
14.669
61.680
1.00
56.88
A


488
NZ
LYS
69
14.926
13.657
61.657
1.00
57.06
A


489
C
LYS
69
12.947
19.472
59.773
1.00
56.39
A


490
O
LYS
69
12.129
19.539
58.854
1.00
56.43
A


491
N
SER
70
13.599
20.539
60.224
1.00
56.49
A


492
CA
SER
70
13.356
21.849
59.627
1.00
56.59
A


493
CB
SER
70
13.750
22.963
60.596
1.00
56.59
A


494
OG
SER
70
15.159
23.074
60.709
1.00
56.59
A


495
C
SER
70
14.163
21.992
58.336
1.00
56.69
A


496
O
SER
70
15.172
21.303
58.149
1.00
56.70
A


497
N
SER
71
13.722
22.883
57.450
1.00
56.67
A


498
CA
SER
71
14.433
23.106
56.193
1.00
56.71
A


499
CB
SER
71
13.802
24.252
55.401
1.00
56.61
A


500
OG
SER
71
12.457
23.959
55.072
1.00
56.71
A


501
C
SER
71
15.888
23.440
56.476
1.00
56.73
A


502
O
SER
71
16.792
22.941
55.805
1.00
56.77
A


503
N
GLU
72
16.107
24.277
57.483
1.00
56.79
A


504
CA
GLU
72
17.450
24.693
57.849
1.00
56.88
A


505
CB
GLU
72
17.402
25.690
59.012
1.00
57.38
A


506
CG
GLU
72
16.430
26.852
58.823
1.00
58.18
A


507
CD
GLU
72
14.973
26.428
58.943
1.00
58.60
A


508
OE1
GLU
72
14.677
25.517
59.756
1.00
59.03
A


509
OE2
GLU
72
14.113
27.010
58.239
1.00
58.90
A


510
C
GLU
72
18.281
23.479
58.246
1.00
56.65
A


511
O
GLU
72
19.474
23.402
57.934
1.00
56.67
A


512
N
GLU
73
17.645
22.534
58.931
1.00
56.31
A


513
CA
GLU
73
18.327
21.321
59.373
1.00
55.99
A


514
CB
GLU
73
17.479
20.596
60.411
1.00
56.34
A


515
CG
GLU
73
18.198
19.447
61.076
1.00
56.94
A


516
CD
GLU
73
17.366
18.786
62.153
1.00
57.34
A


517
OE1
GLU
73
16.262
19.308
62.478
1.00
57.53
A


518
OE2
GLU
73
17.826
17.741
62.678
1.00
57.51
A


519
C
GLU
73
18.623
20.382
58.204
1.00
55.46
A


520
O
GLU
73
19.726
19.846
58.093
1.00
55.44
A


521
N
LEU
74
17.640
20.188
57.332
1.00
54.85
A


522
CA
LEU
74
17.834
19.320
56.179
1.00
54.21
A


523
CB
LEU
74
16.557
19.230
55.341
1.00
54.21
A


524
CG
LEU
74
15.374
18.505
55.982
1.00
54.26
A


525
CD1
LEU
74
14.218
18.545
55.015
1.00
54.12
A


526
CD2
LEU
74
15.741
17.073
56.313
1.00
54.10
A


527
C
LEU
74
18.966
19.869
55.327
1.00
53.75
A


528
O
LEU
74
19.890
19.141
54.975
1.00
53.62
A


529
N
LEU
75
18.917
21.165
55.041
1.00
53.20
A


530
CA
LEU
75
19.934
21.792
54.203
1.00
52.62
A


531
CB
LEU
75
19.693
23.306
54.085
1.00
52.66
A


532
CG
LEU
75
20.678
24.078
53.193
1.00
52.68
A


533
CD1
LEU
75
20.657
23.565
51.765
1.00
52.54
A


534
CD2
LEU
75
20.293
25.542
53.215
1.00
52.73
A


535
C
LEU
75
21.367
21.539
54.654
1.00
52.18
A


536
O
LEU
75
22.246
21.360
53.821
1.00
52.13
A


537
N
GLU
76
21.604
21.498
55.960
1.00
51.70
A


538
CA
GLU
76
22.960
21.288
56.468
1.00
51.14
A


539
CB
GLU
76
23.139
22.025
57.805
1.00
51.69
A


540
CG
GLU
76
22.263
21.456
58.919
1.00
52.39
A


541
CD
GLU
76
22.485
22.120
60.274
1.00
52.91
A


542
OE1
GLU
76
23.297
23.078
60.349
1.00
53.16
A


543
OE2
GLU
76
21.840
21.683
61.267
1.00
53.17
A


544
C
GLU
76
23.295
19.810
56.670
1.00
50.46
A


545
O
GLU
76
24.387
19.481
57.134
1.00
50.48
A


546
N
THR
77
22.376
18.916
56.316
1.00
49.52
A


547
CA
THR
77
22.608
17.482
56.517
1.00
48.55
A


548
CB
THR
77
21.393
16.832
57.210
1.00
48.64
A


549
OG1
THR
77
21.011
17.623
58.341
1.00
48.63
A


550
CG2
THR
77
21.737
15.424
57.678
1.00
48.51
A


551
C
THR
77
22.905
16.709
55.232
1.00
47.86
A


552
O
THR
77
21.999
16.427
54.447
1.00
47.69
A


553
N
LEU
78
24.176
16.369
55.026
1.00
46.95
A


554
CA
LEU
78
24.590
15.635
53.835
1.00
46.11
A


555
CB
LEU
78
26.095
15.337
53.877
1.00
46.00
A


556
CG
LEU
78
27.036
16.539
53.810
1.00
45.95
A


557
CD1
LEU
78
28.458
16.058
53.908
1.00
45.93
A


558
CD2
LEU
78
26.831
17.293
52.524
1.00
45.83
A


559
C
LEU
78
23.825
14.322
53.664
1.00
45.56
A


560
O
LEU
78
23.487
13.936
52.548
1.00
45.41
A


561
N
ASP
79
23.553
13.646
54.775
1.00
44.78
A


562
CA
ASP
79
22.845
12.377
54.739
1.00
44.19
A


563
CB
ASP
79
22.620
11.853
56.160
1.00
44.07
A


564
CG
ASP
79
22.214
10.395
56.183
1.00
43.85
A


565
OD1
ASP
79
21.292
10.057
56.945
1.00
43.81
A


566
OD2
ASP
79
22.827
9.591
55.452
1.00
43.55
A


567
C
ASP
79
21.507
12.534
54.029
1.00
43.87
A


568
O
ASP
79
21.081
11.650
53.287
1.00
43.79
A


569
N
PHE
80
20.847
13.663
54.268
1.00
43.46
A


570
CA
PHE
80
19.559
13.946
53.649
1.00
43.12
A


571
CB
PHE
80
19.080
15.339
54.050
1.00
43.22
A


572
CG
PHE
80
17.830
15.779
53.339
1.00
43.41
A


573
CD1
PHE
80
17.794
17.000
52.664
1.00
43.41
A


574
CD2
PHE
80
16.686
14.984
53.355
1.00
43.40
A


575
CE1
PHE
80
16.633
17.428
52.011
1.00
43.56
A


576
CE2
PHE
80
15.517
15.397
52.707
1.00
43.61
A


577
CZ
PHE
80
15.490
16.624
52.033
1.00
43.59
A


578
C
PHE
80
19.677
13.870
52.131
1.00
42.88
A


579
O
PHE
80
18.904
13.168
51.463
1.00
42.77
A


580
N
TYR
81
20.652
14.592
51.591
1.00
42.41
A


581
CA
TYR
81
20.863
14.605
50.156
1.00
42.04
A


582
CB
TYR
81
21.844
15.722
49.789
1.00
42.05
A


583
CG
TYR
81
21.238
17.071
50.098
1.00
42.06
A


584
CD1
TYR
81
21.532
17.740
51.288
1.00
42.06
A


585
CE1
TYR
81
20.858
18.913
51.637
1.00
42.06
A


586
CD2
TYR
81
20.262
17.615
49.261
1.00
41.96
A


587
CE2
TYR
81
19.587
18.776
49.598
1.00
41.95
A


588
CZ
TYR
81
19.884
19.421
50.785
1.00
42.02
A


589
OH
TYR
81
19.190
20.562
51.121
1.00
42.06
A


590
C
TYR
81
21.316
13.254
49.635
1.00
41.73
A


591
O
TYR
81
21.047
12.914
48.484
1.00
41.56
A


592
N
ARG
82
21.989
12.472
50.476
1.00
41.49
A


593
CA
ARG
82
22.416
11.146
50.043
1.00
41.39
A


594
CB
ARG
82
23.368
10.501
51.053
1.00
41.56
A


595
CG
ARG
82
24.743
11.152
51.094
1.00
41.85
A


596
CD
ARG
82
25.827
10.159
51.476
1.00
41.94
A


597
NE
ARG
82
27.135
10.810
51.497
1.00
42.20
A


598
CZ
ARG
82
27.610
11.489
52.535
1.00
42.16
A


599
NH1
ARG
82
26.889
11.597
53.642
1.00
42.08
A


600
NH2
ARG
82
28.798
12.079
52.456
1.00
42.21
A


601
C
ARG
82
21.189
10.261
49.861
1.00
41.18
A


602
O
ARG
82
21.137
9.456
48.935
1.00
41.19
A


603
N
ASP
83
20.207
10.405
50.745
1.00
40.88
A


604
CA
ASP
83
18.989
9.610
50.631
1.00
40.73
A


605
CB
ASP
83
18.086
9.794
51.848
1.00
40.55
A


606
CG
ASP
83
18.582
9.036
53.055
1.00
40.56
A


607
OD1
ASP
83
19.253
8.002
52.869
1.00
40.49
A


608
OD2
ASP
83
18.289
9.464
54.189
1.00
40.67
A


609
C
ASP
83
18.236
10.017
49.377
1.00
40.62
A


610
O
ASP
83
17.642
9.182
48.709
1.00
40.71
A


611
N
ILE
84
18.259
11.303
49.055
1.00
40.36
A


612
CA
ILE
84
17.582
11.765
47.857
1.00
40.35
A


613
CB
ILE
84
17.556
13.302
47.774
1.00
40.35
A


614
CG2
ILE
84
17.102
13.737
46.387
1.00
40.17
A


615
CG1
ILE
84
16.619
13.860
48.840
1.00
40.41
A


616
CD1
ILE
84
16.633
15.367
48.925
1.00
40.76
A


617
C
ILE
84
18.278
11.238
46.607
1.00
40.24
A


618
O
ILE
84
17.624
10.773
45.669
1.00
40.39
A


619
N
ILE
85
19.605
11.309
46.598
1.00
39.99
A


620
CA
ILE
85
20.393
10.872
45.454
1.00
39.77
A


621
CB
ILE
85
21.885
11.237
45.632
1.00
39.66
A


622
CG2
ILE
85
22.722
10.612
44.517
1.00
39.42
A


623
CG1
ILE
85
22.051
12.762
45.654
1.00
39.65
A


624
CD1
ILE
85
23.433
13.217
46.110
1.00
39.43
A


625
C
ILE
85
20.306
9.377
45.202
1.00
39.75
A


626
O
ILE
85
20.106
8.936
44.067
1.00
39.69
A


627
N
SER
86
20.458
8.600
46.266
1.00
39.72
A


628
CA
SER
86
20.446
7.149
46.152
1.00
39.80
A


629
CB
SER
86
21.688
6.589
46.841
1.00
39.86
A


630
OG
SER
86
22.853
7.094
46.221
1.00
40.14
A


631
C
SER
86
19.209
6.478
46.725
1.00
39.65
A


632
O
SER
86
19.219
5.278
46.986
1.00
39.70
A


633
N
GLY
87
18.148
7.251
46.917
1.00
39.62
A


634
CA
GLY
87
16.922
6.701
47.468
1.00
39.42
A


635
C
GLY
87
16.271
5.701
46.537
1.00
39.31
A


636
O
GLY
87
16.462
5.760
45.323
1.00
39.13
A


637
N
PRO
88
15.490
4.762
47.089
1.00
39.33
A


638
CD
PRO
88
15.214
4.644
48.532
1.00
39.36
A


639
CA
PRO
88
14.787
3.719
46.342
1.00
39.36
A


640
CB
PRO
88
14.417
2.723
47.435
1.00
39.34
A


641
CG
PRO
88
14.090
3.626
48.575
1.00
39.38
A


642
C
PRO
88
13.568
4.264
45.603
1.00
39.39
A


643
O
PRO
88
12.429
3.862
45.851
1.00
39.44
A


644
N
PHE
89
13.814
5.200
44.700
1.00
39.46
A


645
CA
PHE
89
12.755
5.799
43.909
1.00
39.53
A


646
CB
PHE
89
11.944
6.787
44.753
1.00
39.50
A


647
CG
PHE
89
12.782
7.703
45.591
1.00
39.55
A


648
CD1
PHE
89
13.426
8.803
45.020
1.00
39.59
A


649
CD2
PHE
89
12.937
7.465
46.957
1.00
39.49
A


650
CE1
PHE
89
14.211
9.653
45.796
1.00
39.50
A


651
CE2
PHE
89
13.723
8.309
47.745
1.00
39.56
A


652
CZ
PHE
89
14.362
9.409
47.156
1.00
39.51
A


653
C
PHE
89
13.394
6.486
42.721
1.00
39.68
A


654
O
PHE
89
14.564
6.878
42.764
1.00
39.78
A


655
N
GLU
90
12.631
6.623
41.651
1.00
39.59
A


656
CA
GLU
90
13.148
7.230
40.449
1.00
39.72
A


657
CB
GLU
90
12.242
6.863
39.281
1.00
39.81
A


658
CG
GLU
90
12.930
6.874
37.953
1.00
40.17
A


659
CD
GLU
90
12.091
6.233
36.879
1.00
40.25
A


660
OE1
GLU
90
11.068
6.830
36.495
1.00
40.45
A


661
OE2
GLU
90
12.453
5.131
36.424
1.00
40.31
A


662
C
GLU
90
13.265
8.740
40.574
1.00
39.72
A


663
O
GLU
90
12.461
9.383
41.245
1.00
39.73
A


664
N
LYS
91
14.302
9.298
39.963
1.00
39.62
A


665
CA
LYS
91
14.475
10.743
39.958
1.00
39.59
A


666
CB
LYS
91
15.810
11.171
40.594
1.00
39.42
A


667
CG
LYS
91
16.011
10.724
42.039
1.00
39.26
A


668
CD
LYS
91
16.765
9.392
42.103
1.00
38.93
A


669
CE
LYS
91
16.814
8.844
43.521
1.00
38.82
A


670
NZ
LYS
91
17.615
7.592
43.607
1.00
38.74
A


671
C
LYS
91
14.460
11.137
38.487
1.00
39.68
A


672
O
LYS
91
14.992
10.425
37.641
1.00
39.50
A


673
N
LEU
92
13.821
12.254
38.174
1.00
39.83
A


674
CA
LEU
92
13.783
12.718
36.800
1.00
40.17
A


675
CB
LEU
92
12.336
12.747
36.283
1.00
40.40
A


676
CG
LEU
92
12.157
13.210
34.837
1.00
40.65
A


677
CD1
LEU
92
12.849
12.250
33.875
1.00
40.70
A


678
CD2
LEU
92
10.666
13.288
34.530
1.00
40.99
A


679
C
LEU
92
14.390
14.119
36.785
1.00
40.17
A


680
O
LEU
92
13.913
15.010
37.485
1.00
40.31
A


681
N
ILE
93
15.463
14.301
36.022
1.00
40.18
A


682
CA
ILE
93
16.112
15.600
35.945
1.00
40.24
A


683
CB
ILE
93
17.638
15.520
36.226
1.00
40.35
A


684
CG2
ILE
93
18.256
16.909
36.135
1.00
40.08
A


685
CG1
ILE
93
17.904
14.954
37.623
1.00
40.48
A


686
CD1
ILE
93
17.779
13.466
37.716
1.00
40.93
A


687
C
ILE
93
15.920
16.207
34.565
1.00
40.31
A


688
O
ILE
93
16.262
15.596
33.549
1.00
40.16
A


689
N
ARG
94
15.362
17.408
34.523
1.00
40.41
A


690
CA
ARG
94
15.163
18.061
33.244
1.00
40.74
A


691
CB
ARG
94
13.698
18.427
33.008
1.00
41.08
A


692
CG
ARG
94
13.453
18.908
31.576
1.00
41.68
A


693
CD
ARG
94
13.049
20.373
31.509
1.00
42.37
A


694
NE
ARG
94
11.673
20.578
31.942
1.00
42.92
A


695
CZ
ARG
94
11.073
21.762
32.013
1.00
43.32
A


696
NH1
ARG
94
11.724
22.867
31.680
1.00
43.61
A


697
NH2
ARG
94
9.814
21.844
32.423
1.00
43.78
A


698
C
ARG
94
15.991
19.318
33.126
1.00
40.74
A


699
O
ARG
94
15.840
20.255
33.913
1.00
40.65
A


700
N
GLY
95
16.884
19.316
32.145
1.00
40.68
A


701
CA
GLY
95
17.710
20.477
31.890
1.00
40.71
A


702
C
GLY
95
17.156
21.129
30.639
1.00
40.71
A


703
O
GLY
95
16.993
20.470
29.612
1.00
40.76
A


704
N
SER
96
16.822
22.410
30.729
1.00
40.71
A


705
CA
SER
96
16.301
23.142
29.582
1.00
40.80
A


706
CB
SER
96
14.893
23.661
29.860
1.00
40.81
A


707
OG
SER
96
13.985
22.583
29.977
1.00
40.67
A


708
C
SER
96
17.244
24.306
29.336
1.00
40.89
A


709
O
SER
96
17.483
25.122
30.230
1.00
40.93
A


710
N
LYS
97
17.776
24.378
28.124
1.00
40.80
A


711
CA
LYS
97
18.731
25.416
27.786
1.00
40.86
A


712
CB
LYS
97
19.381
25.103
26.433
1.00
40.68
A


713
CG
LYS
97
20.573
24.152
26.517
1.00
40.35
A


714
CD
LYS
97
20.226
22.849
27.248
1.00
40.01
A


715
CE
LYS
97
21.422
21.914
27.291
1.00
39.84
A


716
NZ
LYS
97
21.064
20.548
27.771
1.00
39.74
A


717
C
LYS
97
18.224
26.854
27.789
1.00
40.95
A


718
O
LYS
97
17.078
27.145
27.423
1.00
40.98
A


719
N
ILE
98
19.106
27.743
28.236
1.00
40.96
A


720
CA
ILE
98
18.844
29.173
28.271
1.00
41.07
A


721
CB
ILE
98
19.338
29.790
29.600
1.00
41.05
A


722
CG2
ILE
98
19.259
31.320
29.545
1.00
41.10
A


723
CG1
ILE
98
18.487
29.242
30.755
1.00
40.95
A


724
CD1
ILE
98
18.976
29.640
32.139
1.00
40.81
A


725
C
ILE
98
19.650
29.702
27.089
1.00
41.19
A


726
O
ILE
98
19.202
30.575
26.349
1.00
41.33
A


727
N
ARG
99
20.847
29.154
26.919
1.00
41.28
A


728
CA
ARG
99
21.717
29.509
25.807
1.00
41.49
A


729
CB
ARG
99
23.057
30.052
26.305
1.00
41.83
A


730
CG
ARG
99
22.983
31.464
26.849
1.00
42.32
A


731
CD
ARG
99
24.379
31.981
27.150
1.00
42.87
A


732
NE
ARG
99
24.403
33.442
27.207
1.00
43.34
A


733
CZ
ARG
99
23.935
34.157
28.220
1.00
43.48
A


734
NH1
ARG
99
23.402
33.560
29.277
1.00
43.60
A


735
NH2
ARG
99
23.999
35.474
28.176
1.00
43.71
A


736
C
ARG
99
21.948
28.230
25.004
1.00
41.43
A


737
O
ARG
99
22.027
27.139
25.578
1.00
41.31
A


738
N
GLU
100
22.070
28.370
23.687
1.00
41.29
A


739
CA
GLU
100
22.257
27.220
22.814
1.00
41.18
A


740
CB
GLU
100
22.101
27.635
21.339
1.00
41.57
A


741
CG
GLU
100
22.404
26.510
20.338
1.00
42.34
A


742
CD
GLU
100
21.843
26.775
18.940
1.00
42.90
A


743
OE1
GLU
100
22.205
26.045
17.987
1.00
43.25
A


744
OE2
GLU
100
21.026
27.707
18.790
1.00
43.46
A


745
C
GLU
100
23.567
26.476
23.004
1.00
40.83
A


746
O
GLU
100
24.650
27.054
22.948
1.00
40.71
A


747
N
LEU
101
23.451
25.170
23.217
1.00
40.45
A


748
CA
LEU
101
24.609
24.306
23.400
1.00
40.16
A


749
CB
LEU
101
24.737
23.901
24.866
1.00
40.01
A


750
CG
LEU
101
25.155
24.932
25.909
1.00
39.88
A


751
CD1
LEU
101
24.950
24.343
27.309
1.00
39.74
A


752
CD2
LEU
101
26.617
25.317
25.683
1.00
39.74
A


753
C
LEU
101
24.440
23.043
22.564
1.00
40.00
A


754
O
LEU
101
23.346
22.482
22.499
1.00
39.93
A


755
N
SER
102
25.510
22.592
21.925
1.00
39.88
A


756
CA
SER
102
25.426
21.370
21.145
1.00
39.84
A


757
CB
SER
102
26.591
21.267
20.169
1.00
39.80
A


758
OG
SER
102
27.791
20.981
20.859
1.00
39.80
A


759
C
SER
102
25.511
20.231
22.158
1.00
39.87
A


760
O
SER
102
25.879
20.451
23.317
1.00
39.66
A


761
N
GLY
103
25.161
19.024
21.727
1.00
39.84
A


762
CA
GLY
103
25.224
17.883
22.621
1.00
39.93
A


763
C
GLY
103
26.632
17.705
23.161
1.00
40.16
A


764
O
GLY
103
26.818
17.520
24.361
1.00
39.95
A


765
N
PRO
104
27.652
17.743
22.288
1.00
40.42
A


766
CD
PRO
104
27.560
17.647
20.819
1.00
40.48
A


767
CA
PRO
104
29.041
17.584
22.731
1.00
40.70
A


768
CB
PRO
104
29.826
17.659
21.424
1.00
40.56
A


769
CG
PRO
104
28.893
17.020
20.452
1.00
40.54
A


770
C
PRO
104
29.455
18.676
23.720
1.00
40.97
A


771
O
PRO
104
30.128
18.399
24.714
1.00
40.96
A


772
N
GLU
105
29.048
19.914
23.444
1.00
41.22
A


773
CA
GLU
105
29.388
21.042
24.306
1.00
41.61
A


774
CB
GLU
105
28.878
22.354
23.702
1.00
41.92
A


775
CG
GLU
105
29.707
22.892
22.535
1.00
42.68
A


776
CD
GLU
105
29.090
24.149
21.916
1.00
43.02
A


777
OE1
GLU
105
29.835
24.943
21.305
1.00
43.62
A


778
OE2
GLU
105
27.863
24.345
22.030
1.00
43.13
A


779
C
GLU
105
28.794
20.870
25.698
1.00
41.62
A


780
O
GLU
105
29.488
20.994
26.708
1.00
41.58
A


781
N
TYR
106
27.497
20.600
25.742
1.00
41.60
A


782
CA
TYR
106
26.804
20.411
27.001
1.00
41.79
A


783
CB
TYR
106
25.310
20.229
26.750
1.00
41.53
A


784
CG
TYR
106
24.564
19.746
27.967
1.00
41.25
A


785
CD1
TYR
106
24.233
20.623
29.000
1.00
41.04
A


786
CE1
TYR
106
23.595
20.167
30.155
1.00
40.92
A


787
CD2
TYR
106
24.237
18.396
28.114
1.00
41.14
A


788
CE2
TYR
106
23.602
17.932
29.261
1.00
40.81
A


789
CZ
TYR
106
23.285
18.819
30.273
1.00
40.79
A


790
OH
TYR
106
22.664
18.357
31.404
1.00
40.49
A


791
C
TYR
106
27.332
19.202
27.768
1.00
42.09
A


792
O
TYR
106
27.694
19.306
28.938
1.00
42.10
A


793
N
SER
107
27.386
18.057
27.100
1.00
42.42
A


794
CA
SER
107
27.829
16.823
27.737
1.00
42.92
A


795
CB
SER
107
27.668
15.640
26.774
1.00
42.84
A


796
OG
SER
107
28.508
15.765
25.645
1.00
42.92
A


797
C
SER
107
29.257
16.878
28.254
1.00
43.18
A


798
O
SER
107
29.586
16.220
29.242
1.00
42.97
A


799
N
ARG
108
30.112
17.668
27.615
1.00
43.61
A


800
CA
ARG
108
31.502
17.763
28.027
1.00
44.18
A


801
CB
ARG
108
32.250
18.796
27.185
1.00
44.91
A


802
CG
ARG
108
33.755
18.799
27.407
1.00
46.10
A


803
CD
ARG
108
34.408
19.994
26.733
1.00
47.26
A


804
NE
ARG
108
35.852
20.020
26.944
1.00
48.40
A


805
CZ
ARG
108
36.722
19.304
26.240
1.00
49.05
A


806
NH1
ARG
108
36.292
18.501
25.275
1.00
49.36
A


807
NH2
ARG
108
38.019
19.390
26.501
1.00
49.37
A


808
C
ARG
108
31.607
18.104
29.503
1.00
44.06
A


809
O
ARG
108
32.292
17.405
30.263
1.00
44.00
A


810
N
LYS
109
30.958
19.181
29.940
1.00
44.03
A


811
CA
LYS
109
31.033
19.603
31.331
1.00
44.10
A


812
CB
LYS
109
30.403
20.985
31.507
1.00
44.72
A


813
CG
LYS
109
31.074
22.080
30.693
1.00
45.65
A


814
CD
LYS
109
32.449
22.414
31.248
1.00
46.30
A


815
CE
LYS
109
33.269
23.210
30.245
1.00
46.72
A


816
NZ
LYS
109
34.617
23.551
30.776
1.00
47.36
A


817
C
LYS
109
30.359
18.590
32.248
1.00
43.77
A


818
O
LYS
109
30.894
18.272
33.314
1.00
43.65
A


819
N
VAL
110
29.189
18.088
31.867
1.00
43.35
A


820
CA
VAL
110
28.498
17.111
32.707
1.00
42.90
A


821
CB
VAL
110
27.146
16.668
32.087
1.00
42.83
A


822
CG1
VAL
110
26.512
15.580
32.945
1.00
42.48
A


823
CG2
VAL
110
26.210
17.869
31.980
1.00
42.55
A


824
C
VAL
110
29.373
15.884
32.921
1.00
42.75
A


825
O
VAL
110
29.565
15.434
34.052
1.00
42.30
A


826
N
MET
111
29.914
15.350
31.834
1.00
42.78
A


827
CA
MET
111
30.765
14.176
31.919
1.00
42.98
A


828
CB
MET
111
31.184
13.709
30.523
1.00
43.15
A


829
CG
MET
111
30.008
13.332
29.614
1.00
43.57
A


830
SD
MET
111
30.531
12.486
28.103
1.00
44.26
A


831
CE
MET
111
31.374
13.836
27.227
1.00
44.12
A


832
C
MET
111
32.003
14.449
32.770
1.00
43.03
A


833
O
MET
111
32.434
13.585
33.533
1.00
43.04
A


834
N
GLU
112
32.579
15.644
32.637
1.00
42.98
A


835
CA
GLU
112
33.766
15.999
33.418
1.00
42.93
A


836
CB
GLU
112
34.186
17.449
33.153
1.00
43.27
A


837
CG
GLU
112
34.872
17.649
31.820
1.00
44.06
A


838
CD
GLU
112
35.236
19.101
31.547
1.00
44.44
A


839
OE1
GLU
112
34.995
19.967
32.423
1.00
44.81
A


840
OE2
GLU
112
35.766
19.368
30.444
1.00
44.92
A


841
C
GLU
112
33.473
15.834
34.897
1.00
42.51
A


842
O
GLU
112
34.205
15.157
35.614
1.00
42.29
A


843
N
ASN
113
32.392
16.466
35.339
1.00
42.25
A


844
CA
ASN
113
31.976
16.403
36.732
1.00
42.06
A


845
CB
ASN
113
30.792
17.336
36.953
1.00
42.02
A


846
CG
ASN
113
31.160
18.778
36.713
1.00
42.12
A


847
OD1
ASN
113
32.324
19.080
36.443
1.00
41.97
A


848
ND2
ASN
113
30.184
19.677
36.810
1.00
41.88
A


849
C
ASN
113
31.624
14.992
37.174
1.00
42.00
A


850
O
ASN
113
31.921
14.598
38.309
1.00
41.99
A


851
N
CYS
114
30.992
14.228
36.286
1.00
41.78
A


852
CA
CYS
114
30.626
12.862
36.622
1.00
41.74
A


853
CB
CYS
114
29.749
12.244
35.522
1.00
41.53
A


854
SG
CYS
114
28.039
12.830
35.584
1.00
41.30
A


855
C
CYS
114
31.884
12.037
36.815
1.00
41.88
A


856
O
CYS
114
32.001
11.280
37.775
1.00
41.78
A


857
N
VAL
115
32.836
12.196
35.907
1.00
42.06
A


858
CA
VAL
115
34.075
11.447
35.999
1.00
42.47
A


859
CB
VAL
115
34.954
11.695
34.757
1.00
42.46
A


860
CG1
VAL
115
36.291
10.987
34.909
1.00
42.38
A


861
CG2
VAL
115
34.226
11.195
33.514
1.00
42.49
A


862
C
VAL
115
34.854
11.801
37.268
1.00
42.74
A


863
O
VAL
115
35.393
10.920
37.933
1.00
42.62
A


864
N
ALA
116
34.908
13.088
37.599
1.00
43.18
A


865
CA
ALA
116
35.618
13.535
38.795
1.00
43.74
A


866
CB
ALA
116
35.552
15.063
38.913
1.00
43.60
A


867
C
ALA
116
34.972
12.883
40.015
1.00
44.09
A


868
O
ALA
116
35.653
12.348
40.892
1.00
44.07
A


869
N
HIS
117
33.648
12.929
40.071
1.00
44.53
A


870
CA
HIS
117
32.950
12.315
41.189
1.00
44.95
A


871
CB
HIS
117
31.433
12.446
41.032
1.00
45.11
A


872
CG
HIS
117
30.664
11.576
41.979
1.00
45.31
A


873
CD2
HIS
117
29.919
10.467
41.762
1.00
45.38
A


874
ND1
HIS
117
30.687
11.763
43.346
1.00
45.33
A


875
CE1
HIS
117
29.992
10.803
43.929
1.00
45.43
A


876
NE2
HIS
117
29.516
10.002
42.991
1.00
45.53
A


877
C
HIS
117
33.306
10.838
41.285
1.00
45.24
A


878
O
HIS
117
33.764
10.369
42.323
1.00
45.23
A


879
N
LEU
118
33.101
10.112
40.192
1.00
45.60
A


880
CA
LEU
118
33.368
8.679
40.161
1.00
46.15
A


881
CB
LEU
118
33.093
8.129
38.765
1.00
46.02
A


882
CG
LEU
118
31.624
8.251
38.350
1.00
46.10
A


883
CD1
LEU
118
31.475
7.974
36.859
1.00
46.01
A


884
CD2
LEU
118
30.779
7.286
39.180
1.00
46.11
A


885
C
LEU
118
34.774
8.306
40.610
1.00
46.58
A


886
O
LEU
118
34.943
7.379
41.403
1.00
46.56
A


887
N
LYS
119
35.779
9.010
40.097
1.00
47.09
A


888
CA
LYS
119
37.159
8.740
40.492
1.00
47.62
A


889
CB
LYS
119
38.133
9.573
39.655
1.00
47.78
A


890
CG
LYS
119
38.212
9.155
38.205
1.00
48.15
A


891
CD
LYS
119
39.222
9.999
37.451
1.00
48.55
A


892
CE
LYS
119
39.383
9.510
36.029
1.00
48.80
A


893
NZ
LYS
119
40.542
10.166
35.376
1.00
49.23
A


894
C
LYS
119
37.296
9.109
41.966
1.00
47.88
A


895
O
LYS
119
37.943
8.407
42.737
1.00
48.10
A


896
N
SER
120
36.665
10.212
42.352
1.00
48.12
A


897
CA
SER
120
36.700
10.680
43.731
1.00
48.39
A


898
CB
SER
120
35.782
11.894
43.887
1.00
48.60
A


899
OG
SER
120
35.518
12.166
45.253
1.00
49.22
A


900
C
SER
120
36.283
9.603
44.731
1.00
48.39
A


901
O
SER
120
36.952
9.388
45.745
1.00
48.52
A


902
N
VAL
121
35.180
8.922
44.441
1.00
48.23
A


903
CA
VAL
121
34.667
7.886
45.332
1.00
48.06
A


904
CB
VAL
121
33.126
7.883
45.325
1.00
48.03
A


905
CG1
VAL
121
32.608
9.247
45.771
1.00
48.02
A


906
CG2
VAL
121
32.609
7.555
43.927
1.00
48.00
A


907
C
VAL
121
35.172
6.483
45.000
1.00
47.91
A


908
O
VAL
121
34.633
5.493
45.491
1.00
47.99
A


909
N
GLY
122
36.200
6.403
44.163
1.00
47.73
A


910
CA
GLY
122
36.774
5.116
43.804
1.00
47.44
A


911
C
GLY
122
35.947
4.142
42.981
1.00
47.28
A


912
O
GLY
122
36.156
2.935
43.070
1.00
47.24
A


913
N
THR
123
35.012
4.641
42.178
1.00
47.14
A


914
CA
THR
123
34.200
3.759
41.347
1.00
46.98
A


915
CB
THR
123
32.724
3.717
41.805
1.00
46.91
A


916
OG1
THR
123
32.113
4.994
41.583
1.00
46.66
A


917
CG2
THR
123
32.639
3.353
43.281
1.00
46.84
A


918
C
THR
123
34.238
4.205
39.890
1.00
46.97
A


919
O
THR
123
33.216
4.571
39.306
1.00
46.92
A


920
N
TYR
124
35.429
4.195
39.308
1.00
46.84
A


921
CA
TYR
124
35.564
4.576
37.917
1.00
46.66
A


922
CB
TYR
124
36.176
5.971
37.774
1.00
46.86
A


923
CG
TYR
124
36.098
6.477
36.347
1.00
47.23
A


924
CD1
TYR
124
34.862
6.719
35.747
1.00
47.38
A


925
CE1
TYR
124
34.768
7.134
34.425
1.00
47.54
A


926
CD2
TYR
124
37.251
6.669
35.580
1.00
47.35
A


927
CE2
TYR
124
37.168
7.086
34.245
1.00
47.53
A


928
CZ
TYR
124
35.923
7.315
33.679
1.00
47.66
A


929
OH
TYR
124
35.823
7.719
32.368
1.00
47.86
A


930
C
TYR
124
36.444
3.558
37.227
1.00
46.44
A


931
O
TYR
124
37.627
3.798
36.999
1.00
46.53
A


932
N
GLY
125
35.858
2.409
36.912
1.00
46.10
A


933
CA
GLY
125
36.589
1.354
36.239
1.00
45.74
A


934
C
GLY
125
36.263
1.351
34.761
1.00
45.52
A


935
O
GLY
125
35.666
2.302
34.249
1.00
45.43
A


936
N
ASP
126
36.648
.284
34.068
1.00
45.30
A


937
CA
ASP
126
36.388
.191
32.643
1.00
45.09
A


938
CB
ASP
126
36.925
−1.128
32.079
1.00
45.36
A


939
CG
ASP
126
38.446
−1.241
32.192
1.00
45.63
A


940
OD1
ASP
126
39.137
−.197
32.225
1.00
45.67
A


941
OD2
ASP
126
38.950
−2.382
32.233
1.00
45.82
A


942
C
ASP
126
34.897
.304
32.358
1.00
44.81
A


943
O
ASP
126
34.483
1.018
31.444
1.00
44.67
A


944
N
ALA
127
34.091
−.394
33.152
1.00
44.39
A


945
CA
ALA
127
32.648
−.361
32.967
1.00
44.02
A


946
CB
ALA
127
31.956
−1.222
34.028
1.00
43.92
A


947
C
ALA
127
32.130
1.074
33.024
1.00
43.76
A


948
O
ALA
127
31.348
1.487
32.166
1.00
43.61
A


949
N
GLU
128
32.572
1.843
34.014
1.00
43.55
A


950
CA
GLU
128
32.111
3.224
34.126
1.00
43.62
A


951
CB
GLU
128
32.531
3.845
35.463
1.00
43.55
A


952
CG
GLU
128
31.788
3.279
36.678
1.00
43.55
A


953
CD
GLU
128
32.126
1.824
36.941
1.00
43.58
A


954
OE1
GLU
128
33.327
1.508
37.052
1.00
43.53
A


955
OE2
GLU
128
31.201
.991
37.042
1.00
43.62
A


956
C
GLU
128
32.620
4.086
32.972
1.00
43.60
A


957
O
GLU
128
31.911
4.971
32.500
1.00
43.55
A


958
N
ALA
129
33.844
3.827
32.523
1.00
43.63
A


959
CA
ALA
129
34.422
4.585
31.412
1.00
43.86
A


960
CB
ALA
129
35.885
4.182
31.194
1.00
43.78
A


961
C
ALA
129
33.606
4.302
30.159
1.00
43.88
A


962
O
ALA
129
33.249
5.210
29.415
1.00
43.98
A


963
N
GLU
130
33.305
3.032
29.930
1.00
44.01
A


964
CA
GLU
130
32.517
2.656
28.774
1.00
44.23
A


965
CB
GLU
130
32.422
1.135
28.683
1.00
44.93
A


966
CG
GLU
130
33.741
.489
28.282
1.00
46.16
A


967
CD
GLU
130
33.736
−1.016
28.465
1.00
47.06
A


968
OE1
GLU
130
32.695
−1.650
28.167
1.00
47.65
A


969
OE2
GLU
130
34.778
−1.569
28.896
1.00
47.67
A


970
C
GLU
130
31.128
3.280
28.859
1.00
43.90
A


971
O
GLU
130
30.576
3.718
27.849
1.00
43.72
A


972
N
ALA
131
30.568
3.329
30.065
1.00
43.53
A


973
CA
ALA
131
29.249
3.919
30.254
1.00
43.31
A


974
CB
ALA
131
28.762
3.709
31.688
1.00
43.09
A


975
C
ALA
131
29.308
5.407
29.931
1.00
43.19
A


976
O
ALA
131
28.388
5.952
29.318
1.00
43.03
A


977
N
MET
132
30.389
6.066
30.337
1.00
43.13
A


978
CA
MET
132
30.530
7.492
30.060
1.00
43.23
A


979
CB
MET
132
31.697
8.086
30.849
1.00
43.34
A


980
CG
MET
132
31.472
8.107
32.353
1.00
43.56
A


981
SD
MET
132
29.878
8.840
32.809
1.00
43.96
A


982
CE
MET
132
30.021
10.478
32.064
1.00
43.50
A


983
C
MET
132
30.721
7.754
28.565
1.00
43.26
A


984
O
MET
132
30.315
8.802
28.058
1.00
43.15
A


985
N
GLN
133
31.337
6.806
27.862
1.00
43.25
A


986
CA
GLN
133
31.539
6.956
26.420
1.00
43.48
A


987
CB
GLN
133
32.523
5.903
25.893
1.00
43.88
A


988
CG
GLN
133
32.779
5.983
24.384
1.00
44.76
A


989
CD
GLN
133
33.188
7.381
23.923
1.00
45.33
A


990
OE1
GLN
133
34.118
7.984
24.469
1.00
45.84
A


991
NE2
GLN
133
32.494
7.901
22.913
1.00
45.68
A


992
C
GLN
133
30.180
6.794
25.748
1.00
43.22
A


993
O
GLN
133
29.849
7.509
24.798
1.00
43.19
A


994
N
LYS
134
29.383
5.859
26.257
1.00
42.89
A


995
CA
LYS
134
28.054
5.646
25.704
1.00
42.64
A


996
CB
LYS
134
27.389
4.436
26.360
1.00
42.74
A


997
CG
LYS
134
26.011
4.115
25.805
1.00
42.92
A


998
CD
LYS
134
25.502
2.779
26.345
1.00
43.17
A


999
CE
LYS
134
24.109
2.464
25.830
1.00
43.26
A


1000
NZ
LYS
134
23.665
1.122
26.298
1.00
43.66
A


1001
C
LYS
134
27.250
6.912
25.985
1.00
42.33
A


1002
O
LYS
134
26.435
7.349
25.173
1.00
42.13
A


1003
N
PHE
135
27.496
7.499
27.149
1.00
42.17
A


1004
CA
PHE
135
26.825
8.728
27.543
1.00
41.95
A


1005
CB
PHE
135
27.363
9.173
28.905
1.00
41.89
A


1006
CG
PHE
135
26.648
10.357
29.503
1.00
41.74
A


1007
CD1
PHE
135
25.641
10.173
30.439
1.00
41.59
A


1008
CD2
PHE
135
27.025
11.656
29.170
1.00
41.62
A


1009
CE1
PHE
135
25.021
11.265
31.047
1.00
41.62
A


1010
CE2
PHE
135
26.412
12.752
29.771
1.00
41.54
A


1011
CZ
PHE
135
25.410
12.555
30.713
1.00
41.53
A


1012
C
PHE
135
27.154
9.770
26.465
1.00
41.96
A


1013
O
PHE
135
26.262
10.364
25.858
1.00
41.77
A


1014
N
ALA
136
28.448
9.965
26.223
1.00
42.04
A


1015
CA
ALA
136
28.922
10.931
25.229
1.00
42.27
A


1016
CB
ALA
136
30.444
10.863
25.125
1.00
42.09
A


1017
C
ALA
136
28.307
10.689
23.858
1.00
42.41
A


1018
O
ALA
136
27.848
11.618
23.193
1.00
42.42
A


1019
N
GLU
137
28.303
9.430
23.440
1.00
42.64
A


1020
CA
GLU
137
27.767
9.061
22.141
1.00
43.03
A


1021
CB
GLU
137
27.875
7.550
21.951
1.00
43.75
A


1022
CG
GLU
137
28.505
7.155
20.644
1.00
45.13
A


1023
CD
GLU
137
29.983
7.466
20.611
1.00
45.82
A


1024
OE1
GLU
137
30.723
6.842
21.406
1.00
46.56
A


1025
OE2
GLU
137
30.400
8.329
19.801
1.00
46.16
A


1026
C
GLU
137
26.317
9.488
21.970
1.00
42.71
A


1027
O
GLU
137
25.917
9.950
20.899
1.00
42.80
A


1028
N
ALA
138
25.527
9.332
23.025
1.00
42.31
A


1029
CA
ALA
138
24.116
9.691
22.962
1.00
41.96
A


1030
CB
ALA
138
23.417
9.279
24.254
1.00
41.98
A


1031
C
ALA
138
23.894
11.181
22.707
1.00
41.72
A


1032
O
ALA
138
22.880
11.571
22.128
1.00
41.54
A


1033
N
PHE
139
24.837
12.012
23.140
1.00
41.34
A


1034
CA
PHE
139
24.700
13.455
22.965
1.00
41.15
A


1035
CB
PHE
139
25.381
14.213
24.111
1.00
40.74
A


1036
CG
PHE
139
24.588
14.245
25.387
1.00
40.50
A


1037
CD1
PHE
139
24.687
13.214
26.315
1.00
40.18
A


1038
CD2
PHE
139
23.760
15.329
25.672
1.00
40.30
A


1039
CE1
PHE
139
23.978
13.264
27.512
1.00
40.10
A


1040
CE2
PHE
139
23.042
15.388
26.872
1.00
40.16
A


1041
CZ
PHE
139
23.154
14.354
27.791
1.00
40.04
A


1042
C
PHE
139
25.240
14.010
21.655
1.00
41.14
A


1043
O
PHE
139
24.835
15.085
21.238
1.00
41.08
A


1044
N
LYS
140
26.148
13.289
21.006
1.00
41.32
A


1045
CA
LYS
140
26.745
13.779
19.764
1.00
41.72
A


1046
CB
LYS
140
27.740
12.752
19.212
1.00
41.92
A


1047
CG
LYS
140
28.978
12.587
20.086
1.00
42.34
A


1048
CD
LYS
140
29.990
11.644
19.460
1.00
42.79
A


1049
CE
LYS
140
31.231
11.518
20.342
1.00
43.20
A


1050
NZ
LYS
140
32.244
10.587
19.749
1.00
43.64
A


1051
C
LYS
140
25.807
14.230
18.641
1.00
41.79
A


1052
O
LYS
140
26.063
15.245
17.992
1.00
41.89
A


1053
N
PRO
141
24.714
13.494
18.393
1.00
41.82
A


1054
CD
PRO
141
24.354
12.156
18.903
1.00
41.97
A


1055
CA
PRO
141
23.803
13.904
17.318
1.00
41.80
A


1056
CB
PRO
141
23.155
12.585
16.913
1.00
41.90
A


1057
CG
PRO
141
23.005
11.898
18.229
1.00
41.95
A


1058
C
PRO
141
22.765
14.931
17.748
1.00
41.71
A


1059
O
PRO
141
21.848
15.254
16.995
1.00
41.61
A


1060
N
VAL
142
22.915
15.456
18.955
1.00
41.56
A


1061
CA
VAL
142
21.938
16.397
19.466
1.00
41.56
A


1062
CB
VAL
142
21.417
15.924
20.850
1.00
41.66
A


1063
CG1
VAL
142
20.292
16.833
21.335
1.00
41.67
A


1064
CG2
VAL
142
20.936
14.473
20.755
1.00
41.81
A


1065
C
VAL
142
22.427
17.832
19.606
1.00
41.50
A


1066
O
VAL
142
23.612
18.089
19.826
1.00
41.47
A


1067
N
ASN
143
21.496
18.762
19.447
1.00
41.33
A


1068
CA
ASN
143
21.794
20.168
19.626
1.00
41.38
A


1069
CB
ASN
143
21.789
20.935
18.302
1.00
41.38
A


1070
CG
ASN
143
22.159
22.406
18.491
1.00
41.39
A


1071
OD1
ASN
143
23.181
22.719
19.103
1.00
41.27
A


1072
ND2
ASN
143
21.330
23.306
17.972
1.00
41.22
A


1073
C
ASN
143
20.704
20.702
20.533
1.00
41.38
A


1074
O
ASN
143
19.545
20.281
20.442
1.00
41.22
A


1075
N
PHE
144
21.070
21.615
21.422
1.00
41.46
A


1076
CA
PHE
144
20.097
22.179
22.342
1.00
41.70
A


1077
CB
PHE
144
20.501
21.926
23.801
1.00
41.45
A


1078
CG
PHE
144
20.604
20.471
24.174
1.00
41.40
A


1079
CD1
PHE
144
21.843
19.843
24.239
1.00
41.20
A


1080
CD2
PHE
144
19.458
19.739
24.498
1.00
41.26
A


1081
CE1
PHE
144
21.952
18.504
24.625
1.00
41.32
A


1082
CE2
PHE
144
19.550
18.398
24.884
1.00
41.35
A


1083
CZ
PHE
144
20.798
17.779
24.949
1.00
41.26
A


1084
C
PHE
144
19.880
23.677
22.174
1.00
41.98
A


1085
O
PHE
144
20.526
24.486
22.847
1.00
41.96
A


1086
N
PRO
145
18.991
24.073
21.255
1.00
42.28
A


1087
CD
PRO
145
18.284
23.323
20.200
1.00
42.24
A


1088
CA
PRO
145
18.770
25.512
21.117
1.00
42.29
A


1089
CB
PRO
145
17.942
25.611
19.836
1.00
42.43
A


1090
CG
PRO
145
17.196
24.295
19.807
1.00
42.47
A


1091
C
PRO
145
17.990
25.933
22.367
1.00
42.41
A


1092
O
PRO
145
17.510
25.075
23.124
1.00
42.42
A


1093
N
PRO
146
17.863
27.247
22.616
1.00
42.40
A


1094
CD
PRO
146
18.349
28.400
21.833
1.00
42.37
A


1095
CA
PRO
146
17.126
27.687
23.802
1.00
42.32
A


1096
CB
PRO
146
16.937
29.180
23.554
1.00
42.35
A


1097
CG
PRO
146
18.196
29.542
22.812
1.00
42.35
A


1098
C
PRO
146
15.795
26.957
23.938
1.00
42.26
A


1099
O
PRO
146
15.071
26.794
22.963
1.00
42.31
A


1100
N
GLY
147
15.476
26.517
25.148
1.00
42.22
A


1101
CA
GLY
147
14.221
25.819
25.360
1.00
42.03
A


1102
C
GLY
147
14.317
24.318
25.147
1.00
41.85
A


1103
O
GLY
147
13.484
23.572
25.668
1.00
41.83
A


1104
N
ALA
148
15.312
23.879
24.375
1.00
41.67
A


1105
CA
ALA
148
15.515
22.452
24.116
1.00
41.47
A


1106
CB
ALA
148
16.627
22.240
23.091
1.00
41.48
A


1107
C
ALA
148
15.889
21.814
25.446
1.00
41.43
A


1108
O
ALA
148
16.473
22.472
26.317
1.00
41.33
A


1109
N
SER
149
15.571
20.533
25.608
1.00
41.17
A


1110
CA
SER
149
15.833
19.884
26.881
1.00
40.92
A


1111
CB
SER
149
14.532
19.772
27.668
1.00
40.88
A


1112
OG
SER
149
13.888
21.022
27.774
1.00
41.22
A


1113
C
SER
149
16.467
18.513
26.856
1.00
40.65
A


1114
O
SER
149
16.417
17.788
25.860
1.00
40.68
A


1115
N
VAL
150
17.069
18.171
27.986
1.00
40.32
A


1116
CA
VAL
150
17.665
16.865
28.172
1.00
39.84
A


1117
CB
VAL
150
19.185
16.939
28.436
1.00
39.89
A


1118
CG1
VAL
150
19.479
17.829
29.651
1.00
39.70
A


1119
CG2
VAL
150
19.724
15.533
28.670
1.00
39.82
A


1120
C
VAL
150
16.960
16.328
29.404
1.00
39.59
A


1121
O
VAL
150
16.746
17.058
30.374
1.00
39.45
A


1122
N
PHE
151
16.560
15.063
29.351
1.00
39.30
A


1123
CA
PHE
151
15.891
14.440
30.480
1.00
38.87
A


1124
CB
PHE
151
14.508
13.928
30.077
1.00
39.13
A


1125
CG
PHE
151
13.509
15.015
29.823
1.00
39.25
A


1126
CD1
PHE
151
12.636
15.420
30.826
1.00
39.37
A


1127
CD2
PHE
151
13.438
15.632
28.578
1.00
39.36
A


1128
CE1
PHE
151
11.699
16.425
30.597
1.00
39.52
A


1129
CE2
PHE
151
12.503
16.641
28.336
1.00
39.54
A


1130
CZ
PHE
151
11.634
17.037
29.345
1.00
39.45
A


1131
C
PHE
151
16.724
13.270
30.973
1.00
38.58
A


1132
O
PHE
151
17.062
12.366
30.205
1.00
38.41
A


1133
N
TYR
152
17.076
13.308
32.252
1.00
38.13
A


1134
CA
TYR
152
17.834
12.227
32.857
1.00
37.88
A


1135
CB
TYR
152
18.949
12.746
33.768
1.00
37.88
A


1136
CG
TYR
152
20.021
13.550
33.079
1.00
37.74
A


1137
CD1
TYR
152
19.977
14.945
33.066
1.00
37.66
A


1138
CE1
TYR
152
20.986
15.689
32.447
1.00
37.66
A


1139
CD2
TYR
152
21.092
12.917
32.459
1.00
37.76
A


1140
CE2
TYR
152
22.101
13.644
31.841
1.00
37.59
A


1141
CZ
TYR
152
22.043
15.029
31.841
1.00
37.66
A


1142
OH
TYR
152
23.056
15.750
31.251
1.00
37.80
A


1143
C
TYR
152
16.857
11.465
33.729
1.00
37.79
A


1144
O
TYR
152
16.323
12.020
34.695
1.00
37.55
A


1145
N
ARG
153
16.597
10.210
33.387
1.00
37.54
A


1146
CA
ARG
153
15.711
9.416
34.221
1.00
37.53
A


1147
CB
ARG
153
14.763
8.555
33.384
1.00
37.44
A


1148
CG
ARG
153
13.813
7.728
34.245
1.00
37.32
A


1149
CD
ARG
153
12.827
6.961
33.373
1.00
37.28
A


1150
NE
ARG
153
13.515
6.054
32.462
1.00
37.30
A


1151
CZ
ARG
153
13.946
4.839
32.786
1.00
37.33
A


1152
NH1
ARG
153
13.758
4.361
34.013
1.00
37.43
A


1153
NH2
ARG
153
14.574
4.107
31.878
1.00
37.25
A


1154
C
ARG
153
16.626
8.519
35.030
1.00
37.57
A


1155
O
ARG
153
17.244
7.603
34.487
1.00
37.45
A


1156
N
GLN
154
16.741
8.798
36.321
1.00
37.63
A


1157
CA
GLN
154
17.597
7.988
37.173
1.00
37.90
A


1158
CB
GLN
154
18.270
8.850
38.247
1.00
37.95
A


1159
CG
GLN
154
19.330
8.096
39.051
1.00
38.05
A


1160
CD
GLN
154
19.810
8.881
40.260
1.00
38.19
A


1161
OE1
GLN
154
19.877
10.110
40.226
1.00
38.28
A


1162
NE2
GLN
154
20.153
8.175
41.330
1.00
38.08
A


1163
C
GLN
154
16.766
6.901
37.841
1.00
38.01
A


1164
O
GLN
154
16.024
7.159
38.790
1.00
37.91
A


1165
N
SER
155
16.886
5.688
37.320
1.00
38.27
A


1166
CA
SER
155
16.167
4.545
37.852
1.00
38.62
A


1167
CB
SER
155
16.046
3.470
36.784
1.00
38.57
A


1168
OG
SER
155
15.731
2.229
37.378
1.00
39.38
A


1169
C
SER
155
16.936
3.993
39.042
1.00
38.76
A


1170
O
SER
155
18.162
3.876
39.000
1.00
38.81
A


1171
N
PRO
156
16.227
3.633
40.120
1.00
38.88
A


1172
CD
PRO
156
14.770
3.655
40.339
1.00
38.95
A


1173
CA
PRO
156
16.928
3.098
41.288
1.00
38.90
A


1174
CB
PRO
156
15.835
3.050
42.349
1.00
38.90
A


1175
CG
PRO
156
14.612
2.752
41.539
1.00
38.97
A


1176
C
PRO
156
17.544
1.734
40.996
1.00
38.81
A


1177
O
PRO
156
18.340
1.219
41.782
1.00
38.90
A


1178
N
ASP
157
17.180
1.160
39.854
1.00
38.77
A


1179
CA
ASP
157
17.699
−.144
39.457
1.00
38.75
A


1180
CB
ASP
157
16.699
−.854
38.538
1.00
39.02
A


1181
CG
ASP
157
15.319
−.949
39.150
1.00
39.26
A


1182
OD1
ASP
157
15.225
−1.296
40.345
1.00
39.28
A


1183
OD2
ASP
157
14.326
−.680
38.436
1.00
39.64
A


1184
C
ASP
157
19.065
−.048
38.764
1.00
38.67
A


1185
O
ASP
157
19.506
−.991
38.111
1.00
38.65
A


1186
N
GLY
158
19.721
1.102
38.893
1.00
38.54
A


1187
CA
GLY
158
21.042
1.269
38.312
1.00
38.54
A


1188
C
GLY
158
21.105
1.553
36.826
1.00
38.44
A


1189
O
GLY
158
22.040
1.132
36.143
1.00
38.40
A


1190
N
ILE
159
20.115
2.275
36.321
1.00
38.42
A


1191
CA
ILE
159
20.074
2.619
34.909
1.00
38.43
A


1192
CB
ILE
159
18.963
1.846
34.159
1.00
38.57
A


1193
CG2
ILE
159
18.839
2.368
32.726
1.00
38.58
A


1194
CG1
ILE
159
19.270
.346
34.174
1.00
38.75
A


1195
CD1
ILE
159
18.225
−.505
33.457
1.00
39.00
A


1196
C
ILE
159
19.786
4.099
34.784
1.00
38.38
A


1197
O
ILE
159
19.019
4.663
35.565
1.00
38.19
A


1198
N
LEU
160
20.426
4.731
33.812
1.00
38.40
A


1199
CA
LEU
160
20.214
6.148
33.582
1.00
38.59
A


1200
CB
LEU
160
21.539
6.910
33.673
1.00
38.58
A


1201
CG
LEU
160
21.456
8.411
33.380
1.00
38.62
A


1202
CD1
LEU
160
20.672
9.103
34.482
1.00
38.51
A


1203
CD2
LEU
160
22.867
8.990
33.278
1.00
38.65
A


1204
C
LEU
160
19.633
6.275
32.185
1.00
38.66
A


1205
O
LEU
160
20.296
5.950
31.204
1.00
38.58
A


1206
N
GLY
161
18.384
6.714
32.100
1.00
38.88
A


1207
CA
GLY
161
17.765
6.872
30.799
1.00
39.27
A


1208
C
GLY
161
17.975
8.281
30.278
1.00
39.47
A


1209
O
GLY
161
17.758
9.249
31.004
1.00
39.35
A


1210
N
LEU
162
18.407
8.393
29.027
1.00
39.76
A


1211
CA
LEU
162
18.636
9.694
28.406
1.00
40.16
A


1212
CB
LEU
162
20.042
9.757
27.805
1.00
40.00
A


1213
CG
LEU
162
21.192
9.511
28.786
1.00
39.96
A


1214
CD1
LEU
162
22.527
9.575
28.044
1.00
39.90
A


1215
CD2
LEU
162
21.135
10.546
29.900
1.00
39.77
A


1216
C
LEU
162
17.618
10.018
27.315
1.00
40.55
A


1217
O
LEU
162
17.444
9.253
26.368
1.00
40.35
A


1218
N
SER
163
16.958
11.160
27.464
1.00
41.13
A


1219
CA
SER
163
15.974
11.631
26.497
1.00
41.76
A


1220
CB
SER
163
14.566
11.585
27.086
1.00
41.76
A


1221
OG
SER
163
14.176
10.256
27.345
1.00
42.44
A


1222
C
SER
163
16.298
13.067
26.097
1.00
42.06
A


1223
O
SER
163
16.758
13.863
26.917
1.00
41.82
A


1224
N
PHE
164
16.049
13.386
24.834
1.00
42.54
A


1225
CA
PHE
164
16.306
14.720
24.304
1.00
43.18
A


1226
CB
PHE
164
17.444
14.662
23.294
1.00
43.12
A


1227
CG
PHE
164
18.655
13.945
23.800
1.00
43.17
A


1228
CD1
PHE
164
19.087
12.771
23.194
1.00
43.26
A


1229
CD2
PHE
164
19.362
14.436
24.896
1.00
43.19
A


1230
CE1
PHE
164
20.209
12.095
23.672
1.00
43.22
A


1231
CE2
PHE
164
20.482
13.771
25.383
1.00
43.14
A


1232
CZ
PHE
164
20.908
12.601
24.772
1.00
43.37
A


1233
C
PHE
164
15.049
15.255
23.639
1.00
43.78
A


1234
O
PHE
164
14.379
14.540
22.893
1.00
43.65
A


1235
N
SER
165
14.731
16.513
23.919
1.00
44.59
A


1236
CA
SER
165
13.551
17.150
23.355
1.00
45.52
A


1237
CB
SER
165
12.523
17.403
24.458
1.00
45.60
A


1238
OG
SER
165
11.409
18.123
23.962
1.00
45.95
A


1239
C
SER
165
13.912
18.470
22.678
1.00
46.18
A


1240
O
SER
165
14.866
19.143
23.081
1.00
46.10
A


1241
N
PRO
166
13.160
18.849
21.625
1.00
46.85
A


1242
CD
PRO
166
12.093
18.064
20.982
1.00
46.97
A


1243
CA
PRO
166
13.396
20.098
20.890
1.00
47.32
A


1244
CB
PRO
166
12.488
19.971
19.664
1.00
47.26
A


1245
CG
PRO
166
12.218
18.495
19.556
1.00
47.29
A


1246
C
PRO
166
12.965
21.264
21.766
1.00
47.81
A


1247
O
PRO
166
13.396
22.400
21.573
1.00
47.92
A


1248
N
ASP
167
12.083
20.964
22.714
1.00
48.36
A


1249
CA
ASP
167
11.566
21.955
23.643
1.00
48.95
A


1250
CB
ASP
167
10.156
22.381
23.236
1.00
49.26
A


1251
CG
ASP
167
9.263
21.204
22.919
1.00
49.70
A


1252
OD1
ASP
167
9.256
20.227
23.704
1.00
49.86
A


1253
OD2
ASP
167
8.562
21.260
21.882
1.00
49.98
A


1254
C
ASP
167
11.549
21.378
25.052
1.00
49.23
A


1255
O
ASP
167
12.371
20.520
25.381
1.00
49.23
A


1256
N
THR
168
10.608
21.825
25.878
1.00
49.60
A


1257
CA
THR
168
10.537
21.366
27.259
1.00
50.10
A


1258
CB
THR
168
10.070
22.504
28.192
1.00
50.15
A


1259
OG1
THR
168
8.771
22.947
27.781
1.00
50.34
A


1260
CG2
THR
168
11.047
23.678
28.142
1.00
50.15
A


1261
C
THR
168
9.632
20.159
27.496
1.00
50.41
A


1262
O
THR
168
9.537
19.672
28.621
1.00
50.48
A


1263
N
SER
169
8.970
19.673
26.452
1.00
50.70
A


1264
CA
SER
169
8.081
18.529
26.619
1.00
51.03
A


1265
CB
SER
169
7.083
18.458
25.461
1.00
51.11
A


1266
OG
SER
169
7.740
18.288
24.216
1.00
51.50
A


1267
C
SER
169
8.888
17.234
26.707
1.00
51.10
A


1268
O
SER
169
9.896
17.081
26.013
1.00
51.14
A


1269
N
ILE
170
8.453
16.301
27.551
1.00
51.18
A


1270
CA
ILE
170
9.126
15.021
27.726
1.00
51.37
A


1271
CB
ILE
170
8.594
14.235
28.915
1.00
51.51
A


1272
CG2
ILE
170
9.475
13.025
29.174
1.00
51.59
A


1273
CG1
ILE
170
8.538
15.144
30.150
1.00
51.79
A


1274
CD1
ILE
170
7.897
14.447
31.361
1.00
52.03
A


1275
C
ILE
170
8.967
14.132
26.527
1.00
51.28
A


1276
O
ILE
170
7.856
13.844
26.074
1.00
51.32
A


1277
N
PRO
171
10.076
13.654
25.972
1.00
51.20
A


1278
CD
PRO
171
11.504
13.961
26.221
1.00
51.19
A


1279
CA
PRO
171
9.868
12.800
24.800
1.00
51.10
A


1280
CB
PRO
171
11.266
12.621
24.192
1.00
51.16
A


1281
CG
PRO
171
12.262
13.161
25.172
1.00
51.18
A


1282
C
PRO
171
9.230
11.482
25.188
1.00
51.04
A


1283
O
PRO
171
9.411
11.000
26.316
1.00
50.87
A


1284
N
GLU
172
8.491
10.897
24.253
1.00
50.95
A


1285
CA
GLU
172
7.807
9.624
24.481
1.00
50.86
A


1286
CB
GLU
172
6.855
9.351
23.309
1.00
51.49
A


1287
CG
GLU
172
5.612
10.220
23.369
1.00
52.62
A


1288
CD
GLU
172
4.708
9.847
24.542
1.00
53.33
A


1289
OE1
GLU
172
4.950
8.791
25.189
1.00
53.96
A


1290
OE2
GLU
172
3.737
10.599
24.818
1.00
53.74
A


1291
C
GLU
172
8.801
8.469
24.653
1.00
50.31
A


1292
O
GLU
172
8.588
7.577
25.471
1.00
50.24
A


1293
N
LYS
173
9.895
8.512
23.897
1.00
49.72
A


1294
CA
LYS
173
10.902
7.457
23.987
1.00
49.12
A


1295
CB
LYS
173
11.030
6.736
22.644
1.00
49.50
A


1296
CG
LYS
173
9.745
6.060
22.171
1.00
50.05
A


1297
CD
LYS
173
10.013
5.108
20.993
1.00
50.55
A


1298
CE
LYS
173
8.762
4.310
20.583
1.00
50.91
A


1299
NZ
LYS
173
9.021
3.315
19.486
1.00
51.32
A


1300
C
LYS
173
12.270
7.989
24.405
1.00
48.50
A


1301
O
LYS
173
12.581
9.157
24.174
1.00
48.43
A


1302
N
GLU
174
13.092
7.130
25.003
1.00
47.59
A


1303
CA
GLU
174
14.421
7.554
25.418
1.00
46.78
A


1304
CB
GLU
174
14.842
6.858
26.714
1.00
46.63
A


1305
CG
GLU
174
13.877
7.093
27.860
1.00
46.41
A


1306
CD
GLU
174
14.385
6.543
29.171
1.00
46.39
A


1307
OE1
GLU
174
14.706
5.330
29.226
1.00
46.38
A


1308
OE2
GLU
174
14.457
7.319
30.151
1.00
46.26
A


1309
C
GLU
174
15.416
7.264
24.315
1.00
46.31
A


1310
O
GLU
174
15.264
6.300
23.565
1.00
46.29
A


1311
N
ALA
175
16.426
8.119
24.208
1.00
45.59
A


1312
CA
ALA
175
17.449
7.971
23.185
1.00
44.97
A


1313
CB
ALA
175
18.133
9.315
22.939
1.00
44.96
A


1314
C
ALA
175
18.488
6.912
23.553
1.00
44.55
A


1315
O
ALA
175
19.097
6.301
22.679
1.00
44.28
A


1316
N
ALA
176
18.701
6.701
24.847
1.00
44.10
A


1317
CA
ALA
176
19.678
5.707
25.279
1.00
43.81
A


1318
CB
ALA
176
21.095
6.236
25.069
1.00
43.77
A


1319
C
ALA
176
19.485
5.310
26.729
1.00
43.51
A


1320
O
ALA
176
18.808
6.002
27.492
1.00
43.44
A


1321
N
LEU
177
20.077
4.177
27.093
1.00
43.22
A


1322
CA
LEU
177
20.012
3.657
28.452
1.00
42.98
A


1323
CB
LEU
177
19.184
2.366
28.509
1.00
43.12
A


1324
CG
LEU
177
17.671
2.444
28.282
1.00
43.23
A


1325
CD1
LEU
177
17.081
1.029
28.344
1.00
43.35
A


1326
CD2
LEU
177
17.026
3.324
29.344
1.00
43.13
A


1327
C
LEU
177
21.428
3.354
28.897
1.00
42.67
A


1328
O
LEU
177
22.101
2.514
28.305
1.00
42.82
A


1329
N
ILE
178
21.891
4.046
29.928
1.00
42.22
A


1330
CA
ILE
178
23.233
3.810
30.430
1.00
41.67
A


1331
CB
ILE
178
23.907
5.121
30.894
1.00
41.77
A


1332
CG2
ILE
178
25.384
4.874
31.188
1.00
41.56
A


1333
CG1
ILE
178
23.762
6.197
29.812
1.00
41.63
A


1334
CD1
ILE
178
24.386
5.833
28.476
1.00
41.74
A


1335
C
ILE
178
23.042
2.879
31.614
1.00
41.43
A


1336
O
ILE
178
22.576
3.287
32.683
1.00
41.40
A


1337
N
GLU
179
23.388
1.618
31.411
1.00
40.99
A


1338
CA
GLU
179
23.225
.616
32.448
1.00
40.66
A


1339
CB
GLU
179
22.782
−.695
31.801
1.00
41.14
A


1340
CG
GLU
179
21.535
−.493
30.947
1.00
42.12
A


1341
CD
GLU
179
20.907
−1.784
30.475
1.00
42.79
A


1342
OE1
GLU
179
19.833
−1.715
29.832
1.00
43.53
A


1343
OE2
GLU
179
21.475
−2.865
30.742
1.00
43.30
A


1344
C
GLU
179
24.496
.440
33.263
1.00
40.04
A


1345
O
GLU
179
25.349
−.388
32.951
1.00
39.95
A


1346
N
ASN
180
24.609
1.251
34.306
1.00
39.27
A


1347
CA
ASN
180
25.758
1.226
35.205
1.00
38.73
A


1348
CB
ASN
180
26.945
1.954
34.566
1.00
38.45
A


1349
CG
ASN
180
28.172
1.941
35.451
1.00
38.27
A


1350
OD1
ASN
180
28.282
2.728
36.390
1.00
38.08
A


1351
ND2
ASN
180
29.094
1.030
35.167
1.00
38.09
A


1352
C
ASN
180
25.322
1.920
36.493
1.00
38.31
A


1353
O
ASN
180
24.881
3.071
36.467
1.00
38.24
A


1354
N
LYS
181
25.434
1.218
37.614
1.00
37.95
A


1355
CA
LYS
181
24.994
1.760
38.900
1.00
37.61
A


1356
CB
LYS
181
25.182
.718
40.012
1.00
37.74
A


1357
CG
LYS
181
24.609
1.126
41.380
1.00
37.69
A


1358
CD
LYS
181
23.125
1.471
41.280
1.00
37.99
A


1359
CE
LYS
181
22.459
1.584
42.654
1.00
38.20
A


1360
NZ
LYS
181
20.974
1.744
42.544
1.00
38.41
A


1361
C
LYS
181
25.670
3.062
39.309
1.00
37.43
A


1362
O
LYS
181
24.995
4.061
39.588
1.00
37.26
A


1363
N
ALA
182
26.998
3.053
39.341
1.00
37.18
A


1364
CA
ALA
182
27.757
4.231
39.747
1.00
37.12
A


1365
CB
ALA
182
29.259
3.938
39.679
1.00
37.14
A


1366
C
ALA
182
27.415
5.430
38.885
1.00
36.99
A


1367
O
ALA
182
27.093
6.504
39.394
1.00
37.05
A


1368
N
VAL
183
27.468
5.238
37.574
1.00
36.87
A


1369
CA
VAL
183
27.173
6.319
36.653
1.00
36.72
A


1370
CB
VAL
183
27.500
5.896
35.200
1.00
36.81
A


1371
CG1
VAL
183
27.086
6.986
34.212
1.00
36.71
A


1372
CG2
VAL
183
28.997
5.613
35.082
1.00
36.81
A


1373
C
VAL
183
25.723
6.773
36.749
1.00
36.58
A


1374
O
VAL
183
25.438
7.970
36.636
1.00
36.33
A


1375
N
SER
184
24.808
5.836
37.001
1.00
36.47
A


1376
CA
SER
184
23.394
6.199
37.057
1.00
36.55
A


1377
CB
SER
184
22.513
4.974
37.364
1.00
36.41
A


1378
OG
SER
184
22.561
4.611
38.731
1.00
36.53
A


1379
C
SER
184
23.087
7.326
38.036
1.00
36.59
A


1380
O
SER
184
22.214
8.149
37.767
1.00
36.57
A


1381
N
SER
185
23.794
7.382
39.163
1.00
36.79
A


1382
CA
SER
185
23.545
8.446
40.133
1.00
37.20
A


1383
CB
SER
185
23.538
7.891
41.565
1.00
37.17
A


1384
OG
SER
185
24.832
7.467
41.965
1.00
37.40
A


1385
C
SER
185
24.549
9.603
40.047
1.00
37.45
A


1386
O
SER
185
24.365
10.633
40.696
1.00
37.51
A


1387
N
ALA
186
25.589
9.440
39.234
1.00
37.65
A


1388
CA
ALA
186
26.622
10.469
39.097
1.00
37.89
A


1389
CB
ALA
186
27.687
10.009
38.106
1.00
37.74
A


1390
C
ALA
186
26.083
11.833
38.681
1.00
37.97
A


1391
O
ALA
186
26.485
12.861
39.229
1.00
38.04
A


1392
N
VAL
187
25.171
11.850
37.717
1.00
38.08
A


1393
CA
VAL
187
24.621
13.111
37.252
1.00
38.22
A


1394
CB
VAL
187
23.591
12.892
36.118
1.00
38.21
A


1395
CG1
VAL
187
23.066
14.230
35.629
1.00
38.05
A


1396
CG2
VAL
187
24.235
12.126
34.973
1.00
37.92
A


1397
C
VAL
187
23.965
13.884
38.385
1.00
38.42
A


1398
O
VAL
187
24.307
15.040
38.632
1.00
38.34
A


1399
N
LEU
188
23.030
13.248
39.086
1.00
38.62
A


1400
CA
LEU
188
22.339
13.916
40.181
1.00
38.82
A


1401
CB
LEU
188
21.223
13.024
40.739
1.00
38.68
A


1402
CG
LEU
188
20.362
13.651
41.840
1.00
38.63
A


1403
CD1
LEU
188
19.699
14.929
41.332
1.00
38.43
A


1404
CD2
LEU
188
19.311
12.642
42.285
1.00
38.52
A


1405
C
LEU
188
23.320
14.288
41.293
1.00
39.05
A


1406
O
LEU
188
23.174
15.321
41.936
1.00
39.02
A


1407
N
GLU
189
24.316
13.439
41.512
1.00
39.35
A


1408
CA
GLU
189
25.335
13.690
42.524
1.00
39.94
A


1409
CB
GLU
189
26.382
12.573
42.487
1.00
40.24
A


1410
CG
GLU
189
27.439
12.639
43.569
1.00
41.00
A


1411
CD
GLU
189
26.921
12.218
44.942
1.00
41.45
A


1412
OE1
GLU
189
26.265
11.155
45.041
1.00
41.52
A


1413
OE2
GLU
189
27.185
12.943
45.927
1.00
41.70
A


1414
C
GLU
189
26.021
15.041
42.271
1.00
40.13
A


1415
O
GLU
189
26.260
15.808
43.204
1.00
39.98
A


1416
N
THR
190
26.320
15.335
41.006
1.00
40.31
A


1417
CA
THR
190
26.992
16.582
40.661
1.00
40.75
A


1418
CB
THR
190
27.578
16.539
39.233
1.00
40.62
A


1419
OG1
THR
190
26.515
16.440
38.276
1.00
40.54
A


1420
CG2
THR
190
28.510
15.345
39.077
1.00
40.51
A


1421
C
THR
190
26.053
17.775
40.761
1.00
41.14
A


1422
O
THR
190
26.474
18.917
40.617
1.00
41.18
A


1423
N
MET
191
24.780
17.511
41.016
1.00
41.50
A


1424
CA
MET
191
23.812
18.591
41.125
1.00
42.08
A


1425
CB
MET
191
22.542
18.231
40.356
1.00
42.19
A


1426
CG
MET
191
22.790
17.885
38.900
1.00
42.52
A


1427
SD
MET
191
21.284
17.304
38.120
1.00
42.98
A


1428
CE
MET
191
21.674
17.510
36.378
1.00
43.11
A


1429
C
MET
191
23.460
18.926
42.573
1.00
42.40
A


1430
O
MET
191
23.313
20.099
42.920
1.00
42.43
A


1431
N
ILE
192
23.330
17.905
43.415
1.00
42.74
A


1432
CA
ILE
192
22.972
18.124
44.815
1.00
43.28
A


1433
CB
ILE
192
21.499
17.754
45.064
1.00
43.18
A


1434
CG2
ILE
192
20.596
18.639
44.219
1.00
43.15
A


1435
CG1
ILE
192
21.269
16.276
44.731
1.00
43.06
A


1436
CD1
ILE
192
19.848
15.798
45.004
1.00
42.93
A


1437
C
ILE
192
23.831
17.349
45.809
1.00
43.79
A


1438
O
ILE
192
23.538
17.324
47.004
1.00
43.59
A


1439
N
GLY
193
24.882
16.710
45.310
1.00
44.53
A


1440
CA
GLY
193
25.765
15.951
46.173
1.00
45.57
A


1441
C
GLY
193
26.611
16.876
47.032
1.00
46.48
A


1442
O
GLY
193
26.524
18.104
46.910
1.00
46.38
A


1443
N
GLU
194
27.437
16.290
47.893
1.00
47.30
A


1444
CA
GLU
194
28.286
17.064
48.795
1.00
48.24
A


1445
CB
GLU
194
29.124
16.130
49.672
1.00
48.33
A


1446
CG
GLU
194
30.122
16.877
50.545
1.00
48.74
A


1447
CD
GLU
194
31.022
15.953
51.341
1.00
49.05
A


1448
OE1
GLU
194
31.022
14.731
51.068
1.00
49.21
A


1449
OE2
GLU
194
31.739
16.452
52.235
1.00
49.15
A


1450
C
GLU
194
29.221
18.018
48.064
1.00
48.72
A


1451
O
GLU
194
29.404
19.168
48.487
1.00
48.83
A


1452
N
HIS
195
29.820
17.542
46.976
1.00
49.39
A


1453
CA
HIS
195
30.745
18.365
46.214
1.00
50.10
A


1454
CB
HIS
195
31.890
17.516
45.694
1.00
50.52
A


1455
CG
HIS
195
32.670
16.884
46.798
1.00
50.96
A


1456
CD2
HIS
195
33.358
17.433
47.828
1.00
51.16
A


1457
ND1
HIS
195
32.695
15.523
46.999
1.00
51.15
A


1458
CE1
HIS
195
33.367
15.256
48.107
1.00
51.27
A


1459
NE2
HIS
195
33.781
16.398
48.628
1.00
51.39
A


1460
C
HIS
195
30.137
19.185
45.105
1.00
50.36
A


1461
O
HIS
195
30.821
19.604
44.172
1.00
50.51
A


1462
N
ALA
196
28.834
19.412
45.213
1.00
50.55
A


1463
CA
ALA
196
28.152
20.260
44.251
1.00
50.75
A


1464
CB
ALA
196
26.732
19.779
43.998
1.00
50.64
A


1465
C
ALA
196
28.135
21.636
44.926
1.00
50.88
A


1466
O
ALA
196
27.440
21.850
45.924
1.00
50.94
A


1467
N
VAL
197
28.899
22.570
44.377
1.00
51.05
A


1468
CA
VAL
197
28.957
23.906
44.954
1.00
51.23
A


1469
CB
VAL
197
30.044
24.754
44.263
1.00
51.40
A


1470
CG1
VAL
197
30.208
26.092
44.990
1.00
51.42
A


1471
CG2
VAL
197
31.362
23.995
44.264
1.00
51.45
A


1472
C
VAL
197
27.626
24.658
44.920
1.00
51.21
A


1473
O
VAL
197
27.168
25.152
45.958
1.00
51.19
A


1474
N
SER
198
27.012
24.751
43.741
1.00
51.08
A


1475
CA
SER
198
25.734
25.444
43.597
1.00
51.00
A


1476
CB
SER
198
25.172
25.243
42.190
1.00
51.05
A


1477
OG
SER
198
26.092
25.680
41.209
1.00
51.19
A


1478
C
SER
198
24.737
24.911
44.615
1.00
50.92
A


1479
O
SER
198
24.475
23.715
44.673
1.00
50.96
A


1480
N
PRO
199
24.162
25.802
45.433
1.00
50.81
A


1481
CD
PRO
199
24.663
27.150
45.771
1.00
50.78
A


1482
CA
PRO
199
23.199
25.347
46.436
1.00
50.69
A


1483
CB
PRO
199
23.581
26.178
47.649
1.00
50.77
A


1484
CG
PRO
199
23.840
27.514
47.010
1.00
50.73
A


1485
C
PRO
199
21.729
25.538
46.083
1.00
50.57
A


1486
O
PRO
199
20.871
25.108
46.845
1.00
50.55
A


1487
N
ASP
200
21.427
26.163
44.945
1.00
50.43
A


1488
CA
ASP
200
20.029
26.416
44.601
1.00
50.41
A


1489
CB
ASP
200
19.900
27.138
43.241
1.00
50.39
A


1490
CG
ASP
200
20.503
26.363
42.078
1.00
50.36
A


1491
OD1
ASP
200
21.013
25.231
42.272
1.00
50.44
A


1492
OD2
ASP
200
20.469
26.895
40.945
1.00
50.25
A


1493
C
ASP
200
19.117
25.193
44.653
1.00
50.34
A


1494
O
ASP
200
18.030
25.260
45.232
1.00
50.33
A


1495
N
LEU
201
19.550
24.081
44.073
1.00
50.31
A


1496
CA
LEU
201
18.737
22.870
44.083
1.00
50.30
A


1497
CB
LEU
201
19.338
21.825
43.140
1.00
50.26
A


1498
CG
LEU
201
18.675
21.571
41.782
1.00
50.37
A


1499
CD1
LEU
201
18.002
22.802
41.250
1.00
50.33
A


1500
CD2
LEU
201
19.740
21.082
40.827
1.00
50.15
A


1501
C
LEU
201
18.649
22.320
45.505
1.00
50.30
A


1502
O
LEU
201
17.581
21.896
45.947
1.00
50.16
A


1503
N
LYS
202
19.764
22.339
46.226
1.00
50.41
A


1504
CA
LYS
202
19.757
21.839
47.594
1.00
50.68
A


1505
CB
LYS
202
21.153
21.899
48.212
1.00
50.62
A


1506
CG
LYS
202
22.165
20.990
47.549
1.00
50.62
A


1507
CD
LYS
202
23.545
21.212
48.135
1.00
50.63
A


1508
CE
LYS
202
24.597
20.387
47.416
1.00
50.71
A


1509
NZ
LYS
202
25.982
20.706
47.872
1.00
50.72
A


1510
C
LYS
202
18.795
22.657
48.442
1.00
50.90
A


1511
O
LYS
202
18.030
22.102
49.227
1.00
50.78
A


1512
N
ARG
203
18.825
23.973
48.273
1.00
51.26
A


1513
CA
ARG
203
17.953
24.848
49.042
1.00
51.71
A


1514
CB
ARG
203
18.305
26.315
48.792
1.00
52.21
A


1515
CG
ARG
203
19.695
26.687
49.302
1.00
53.05
A


1516
CD
ARG
203
20.058
28.139
49.001
1.00
53.78
A


1517
NE
ARG
203
21.449
28.417
49.352
1.00
54.50
A


1518
CZ
ARG
203
22.082
29.556
49.091
1.00
54.85
A


1519
NH1
ARG
203
21.457
30.554
48.470
1.00
55.02
A


1520
NH2
ARG
203
23.356
29.690
49.444
1.00
55.05
A


1521
C
ARG
203
16.483
24.595
48.738
1.00
51.75
A


1522
O
ARG
203
15.657
24.598
49.647
1.00
51.66
A


1523
N
CYS
204
16.149
24.379
47.469
1.00
51.79
A


1524
CA
CYS
204
14.761
24.105
47.112
1.00
51.94
A


1525
CB
CYS
204
14.626
23.899
45.607
1.00
52.01
A


1526
SG
CYS
204
14.550
25.400
44.646
1.00
52.24
A


1527
C
CYS
204
14.276
22.847
47.820
1.00
52.00
A


1528
O
CYS
204
13.200
22.833
48.413
1.00
51.91
A


1529
N
LEU
205
15.087
21.794
47.739
1.00
52.10
A


1530
CA
LEU
205
14.769
20.507
48.347
1.00
52.26
A


1531
CB
LEU
205
15.884
19.495
48.065
1.00
51.94
A


1532
CG
LEU
205
15.926
18.961
46.632
1.00
51.67
A


1533
CD1
LEU
205
17.242
18.276
46.391
1.00
51.47
A


1534
CD2
LEU
205
14.782
18.004
46.408
1.00
51.46
A


1535
C
LEU
205
14.549
20.632
49.848
1.00
52.57
A


1536
O
LEU
205
13.590
20.082
50.387
1.00
52.54
A


1537
N
ALA
206
15.429
21.367
50.516
1.00
52.95
A


1538
CA
ALA
206
15.311
21.549
51.956
1.00
53.42
A


1539
CB
ALA
206
16.546
22.241
52.501
1.00
53.35
A


1540
C
ALA
206
14.064
22.357
52.294
1.00
53.82
A


1541
O
ALA
206
13.402
22.091
53.295
1.00
53.77
A


1542
N
ALA
207
13.732
23.322
51.444
1.00
54.28
A


1543
CA
ALA
207
12.568
24.165
51.679
1.00
54.86
A


1544
CB
ALA
207
12.703
25.463
50.894
1.00
54.81
A


1545
C
ALA
207
11.254
23.482
51.330
1.00
55.29
A


1546
O
ALA
207
10.261
23.645
52.032
1.00
55.37
A


1547
N
ARG
208
11.257
22.692
50.266
1.00
55.79
A


1548
CA
ARG
208
10.044
22.023
49.813
1.00
56.35
A


1549
CB
ARG
208
10.118
21.775
48.308
1.00
56.32
A


1550
CG
ARG
208
10.421
23.006
47.500
1.00
56.51
A


1551
CD
ARG
208
10.328
22.726
46.018
1.00
56.57
A


1552
NE
ARG
208
9.498
23.740
45.392
1.00
56.83
A


1553
CZ
ARG
208
8.566
23.498
44.487
1.00
56.83
A


1554
NH1
ARG
208
8.326
22.264
44.071
1.00
56.75
A


1555
NH2
ARG
208
7.844
24.509
44.029
1.00
57.04
A


1556
C
ARG
208
9.652
20.711
50.475
1.00
56.77
A


1557
O
ARG
208
8.472
20.463
50.682
1.00
56.73
A


1558
N
LEU
209
10.628
19.868
50.789
1.00
57.33
A


1559
CA
LEU
209
10.340
18.561
51.371
1.00
57.96
A


1560
CB
LEU
209
11.627
17.731
51.446
1.00
57.90
A


1561
CG
LEU
209
11.988
16.846
50.241
1.00
57.92
A


1562
CD1
LEU
209
10.950
15.745
50.075
1.00
57.91
A


1563
CD2
LEU
209
12.082
17.681
48.988
1.00
57.85
A


1564
C
LEU
209
9.626
18.478
52.720
1.00
58.48
A


1565
O
LEU
209
8.736
17.651
52.891
1.00
58.48
A


1566
N
PRO
210
10.006
19.321
53.693
1.00
59.04
A


1567
CD
PRO
210
11.051
20.359
53.644
1.00
59.19
A


1568
CA
PRO
210
9.373
19.289
55.017
1.00
59.61
A


1569
CB
PRO
210
9.895
20.561
55.671
1.00
59.46
A


1570
CG
PRO
210
11.277
20.653
55.115
1.00
59.29
A


1571
C
PRO
210
7.848
19.211
55.011
1.00
60.18
A


1572
O
PRO
210
7.261
18.404
55.731
1.00
60.18
A


1573
N
ALA
211
7.208
20.043
54.197
1.00
60.93
A


1574
CA
ALA
211
5.752
20.055
54.118
1.00
61.74
A


1575
CB
ALA
211
5.293
21.111
53.126
1.00
61.76
A


1576
C
ALA
211
5.216
18.690
53.707
1.00
62.31
A


1577
O
ALA
211
4.278
18.174
54.318
1.00
62.36
A


1578
N
LEU
212
5.815
18.112
52.670
1.00
63.00
A


1579
CA
LEU
212
5.400
16.805
52.169
1.00
63.72
A


1580
CB
LEU
212
6.117
16.493
50.854
1.00
63.70
A


1581
CG
LEU
212
5.865
17.446
49.687
1.00
63.70
A


1582
CD1
LEU
212
6.695
17.016
48.492
1.00
63.67
A


1583
CD2
LEU
212
4.386
17.446
49.338
1.00
63.66
A


1584
C
LEU
212
5.680
15.694
53.175
1.00
64.26
A


1585
O
LEU
212
4.844
14.819
53.400
1.00
64.29
A


1586
N
LEU
213
6.860
15.734
53.781
1.00
64.95
A


1587
CA
LEU
213
7.242
14.724
54.756
1.00
65.72
A


1588
CB
LEU
213
8.704
14.917
55.171
1.00
65.66
A


1589
CG
LEU
213
9.750
14.821
54.056
1.00
65.62
A


1590
CD1
LEU
213
11.138
14.973
54.653
1.00
65.58
A


1591
CD2
LEU
213
9.620
13.486
53.337
1.00
65.62
A


1592
C
LEU
213
6.348
14.750
55.991
1.00
66.31
A


1593
O
LEU
213
6.182
13.732
56.661
1.00
66.41
A


1594
N
ASN
214
5.767
15.908
56.290
1.00
67.00
A


1595
CA
ASN
214
4.900
16.033
57.457
1.00
67.73
A


1596
CB
ASN
214
4.786
17.497
57.883
1.00
67.80
A


1597
CG
ASN
214
5.957
17.939
58.733
1.00
68.01
A


1598
OD1
ASN
214
6.342
17.245
59.676
1.00
68.12
A


1599
ND2
ASN
214
6.526
19.099
58.415
1.00
68.17
A


1600
C
ASN
214
3.508
15.439
57.273
1.00
68.14
A


1601
O
ASN
214
2.852
15.078
58.249
1.00
68.29
A


1602
N
GLU
215
3.055
15.336
56.029
1.00
68.60
A


1603
CA
GLU
215
1.743
14.765
55.751
1.00
69.03
A


1604
CB
GLU
215
.833
15.809
55.097
1.00
69.33
A


1605
CG
GLU
215
1.427
16.486
53.877
1.00
69.76
A


1606
CD
GLU
215
.434
17.403
53.188
1.00
70.11
A


1607
OE1
GLU
215
−.103
18.312
53.865
1.00
70.24
A


1608
OE2
GLU
215
.193
17.212
51.972
1.00
70.24
A


1609
C
GLU
215
1.853
13.531
54.856
1.00
69.16
A


1610
OT1
GLU
215
1.410
13.597
53.686
1.00
69.27
A


1611
OT2
GLU
215
2.391
12.510
55.340
1.00
69.25
A


1612
CB
SER
4
13.836
−22.661
42.351
1.00
61.80
B


1613
OG
SER
4
13.171
−21.446
42.663
1.00
61.99
B


1614
C
SER
4
15.637
−22.063
43.963
1.00
61.58
B


1615
O
SER
4
14.784
−22.189
44.844
1.00
61.68
B


1616
N
SER
4
16.030
−23.817
42.279
1.00
61.77
B


1617
CA
SER
4
15.348
−22.517
42.541
1.00
61.72
B


1618
N
ILE
5
16.847
−21.548
44.176
1.00
61.27
B


1619
CA
ILE
5
17.287
−21.062
45.481
1.00
60.86
B


1620
CB
ILE
5
16.681
−19.642
45.772
1.00
60.98
B


1621
CG2
ILE
5
15.161
−19.701
45.822
1.00
61.01
B


1622
CG1
ILE
5
17.242
−19.080
47.078
1.00
60.98
B


1623
CD1
ILE
5
16.423
−19.415
48.290
1.00
61.06
B


1624
C
ILE
5
16.992
−22.049
46.619
1.00
60.46
B


1625
O
ILE
5
15.847
−22.254
47.032
1.00
60.42
B


1626
N
THR
6
18.060
−22.663
47.116
1.00
59.93
B


1627
CA
THR
6
17.977
−23.649
48.182
1.00
59.38
B


1628
CB
THR
6
18.791
−24.895
47.824
1.00
59.37
B


1629
OG1
THR
6
20.152
−24.511
47.588
1.00
59.31
B


1630
CG2
THR
6
18.235
−25.557
46.581
1.00
59.31
B


1631
C
THR
6
18.530
−23.116
49.497
1.00
58.99
B


1632
O
THR
6
19.154
−22.057
49.540
1.00
58.84
B


1633
N
ALA
7
18.301
−23.867
50.569
1.00
58.52
B


1634
CA
ALA
7
18.805
−23.488
51.874
1.00
58.11
B


1635
CB
ALA
7
18.091
−24.278
52.966
1.00
58.10
B


1636
C
ALA
7
20.291
−23.824
51.861
1.00
57.82
B


1637
O
ALA
7
20.740
−24.647
51.063
1.00
57.74
B


1638
N
ILE
8
21.055
−23.172
52.726
1.00
57.49
B


1639
CA
ILE
8
22.484
−23.426
52.807
1.00
57.18
B


1640
CB
ILE
8
23.315
−22.263
52.234
1.00
57.11
B


1641
CG2
ILE
8
24.796
−22.544
52.429
1.00
57.06
B


1642
CG1
ILE
8
23.003
−22.070
50.750
1.00
57.03
B


1643
CD1
ILE
8
23.673
−20.848
50.148
1.00
57.04
B


1644
C
ILE
8
22.861
−23.583
54.263
1.00
57.06
B


1645
O
ILE
8
22.430
−22.803
55.113
1.00
56.94
B


1646
N
THR
9
23.667
−24.594
54.549
1.00
56.98
B


1647
CA
THR
9
24.107
−24.828
55.912
1.00
56.95
B


1648
CB
THR
9
23.763
−26.256
56.374
1.00
56.98
B


1649
OG1
THR
9
22.338
−26.427
56.349
1.00
57.05
B


1650
CG2
THR
9
24.273
−26.498
57.799
1.00
56.95
B


1651
C
THR
9
25.606
−24.611
55.993
1.00
56.90
B


1652
O
THR
9
26.365
−25.122
55.172
1.00
56.86
B


1653
N
VAL
10
26.021
−23.823
56.974
1.00
56.82
B


1654
CA
VAL
10
27.431
−23.534
57.178
1.00
56.83
B


1655
CB
VAL
10
27.713
−22.017
57.053
1.00
56.70
B


1656
CG1
VAL
10
29.183
−21.726
57.328
1.00
56.57
B


1657
CG2
VAL
10
27.325
−21.535
55.669
1.00
56.63
B


1658
C
VAL
10
27.784
−24.000
58.582
1.00
56.92
B


1659
O
VAL
10
27.317
−23.428
59.565
1.00
56.96
B


1660
N
GLU
11
28.588
−25.053
58.675
1.00
57.03
B


1661
CA
GLU
11
28.989
−25.578
59.978
1.00
57.08
B


1662
CB
GLU
11
29.940
−24.596
60.663
1.00
57.41
B


1663
CG
GLU
11
31.291
−24.455
59.983
1.00
57.98
B


1664
CD
GLU
11
32.137
−25.704
60.119
1.00
58.33
B


1665
OE1
GLU
11
32.355
−26.142
61.269
1.00
58.58
B


1666
OE2
GLU
11
32.585
−26.246
59.084
1.00
58.64
B


1667
C
GLU
11
27.785
−25.833
60.879
1.00
56.87
B


1668
O
GLU
11
27.719
−25.335
62.005
1.00
56.92
B


1669
N
ASN
12
26.829
−26.603
60.374
1.00
56.59
B


1670
CA
ASN
12
25.631
−26.936
61.135
1.00
56.21
B


1671
CB
ASN
12
26.020
−27.661
62.428
1.00
56.68
B


1672
CG
ASN
12
26.858
−28.903
62.165
1.00
57.06
B


1673
OD1
ASN
12
26.479
−29.765
61.364
1.00
57.33
B


1674
ND2
ASN
12
28.003
−29.000
62.834
1.00
57.28
B


1675
C
ASN
12
24.733
−25.738
61.449
1.00
55.73
B


1676
O
ASN
12
23.795
−25.854
62.234
1.00
55.69
B


1677
N
LEU
13
25.027
−24.592
60.840
1.00
55.07
B


1678
CA
LEU
13
24.212
−23.398
61.026
1.00
54.34
B


1679
CB
LEU
13
25.093
−22.165
61.255
1.00
54.30
B


1680
CG
LEU
13
25.882
−22.163
62.568
1.00
54.24
B


1681
CD1
LEU
13
26.883
−21.027
62.565
1.00
54.22
B


1682
CD2
LEU
13
24.925
−22.033
63.745
1.00
54.27
B


1683
C
LEU
13
23.398
−23.256
59.744
1.00
53.90
B


1684
O
LEU
13
23.912
−22.871
58.694
1.00
53.87
B


1685
N
GLU
14
22.122
−23.589
59.839
1.00
53.26
B


1686
CA
GLU
14
21.244
−23.546
58.683
1.00
52.74
B


1687
CB
GLU
14
20.074
−24.504
58.910
1.00
53.34
B


1688
CG
GLU
14
18.709
−23.907
58.609
1.00
54.36
B


1689
CD
GLU
14
17.572
−24.812
59.041
1.00
54.97
B


1690
OE1
GLU
14
17.799
−26.045
59.135
1.00
55.41
B


1691
OE2
GLU
14
16.455
−24.288
59.276
1.00
55.26
B


1692
C
GLU
14
20.707
−22.166
58.320
1.00
51.88
B


1693
O
GLU
14
20.326
−21.376
59.188
1.00
51.76
B


1694
N
TYR
15
20.672
−21.900
57.016
1.00
50.85
B


1695
CA
TYR
15
20.145
−20.658
56.469
1.00
49.85
B


1696
CB
TYR
15
21.212
−19.904
55.672
1.00
49.70
B


1697
CG
TYR
15
22.216
−19.181
56.531
1.00
49.56
B


1698
CD1
TYR
15
23.274
−19.862
57.130
1.00
49.49
B


1699
CE1
TYR
15
24.177
−19.197
57.954
1.00
49.40
B


1700
CD2
TYR
15
22.086
−17.814
56.776
1.00
49.56
B


1701
CE2
TYR
15
22.976
−17.144
57.594
1.00
49.37
B


1702
CZ
TYR
15
24.020
−17.836
58.180
1.00
49.43
B


1703
OH
TYR
15
24.898
−17.155
58.991
1.00
49.23
B


1704
C
TYR
15
18.998
−21.040
55.542
1.00
49.19
B


1705
O
TYR
15
19.220
−21.455
54.409
1.00
49.13
B


1706
N
PRO
16
17.754
−20.927
56.020
1.00
48.65
B


1707
CD
PRO
16
17.283
−20.460
57.334
1.00
48.52
B


1708
CA
PRO
16
16.626
−21.286
55.155
1.00
48.15
B


1709
CB
PRO
16
15.404
−20.973
56.023
1.00
48.28
B


1710
CG
PRO
16
15.914
−19.956
57.002
1.00
48.46
B


1711
C
PRO
16
16.638
−20.517
53.836
1.00
47.65
B


1712
O
PRO
16
17.183
−19.413
53.745
1.00
47.40
B


1713
N
ALA
17
16.034
−21.120
52.819
1.00
47.10
B


1714
CA
ALA
17
15.980
−20.533
51.488
1.00
46.55
B


1715
CB
ALA
17
15.310
−21.513
50.523
1.00
46.52
B


1716
C
ALA
17
15.227
−19.214
51.495
1.00
46.25
B


1717
O
ALA
17
15.556
−18.293
50.745
1.00
46.11
B


1718
N
VAL
18
14.219
−19.118
52.349
1.00
45.82
B


1719
CA
VAL
18
13.421
−17.906
52.421
1.00
45.55
B


1720
CB
VAL
18
12.079
−18.095
51.659
1.00
45.68
B


1721
CG1
VAL
18
11.269
−19.207
52.300
1.00
45.77
B


1722
CG2
VAL
18
11.279
−16.800
51.644
1.00
45.93
B


1723
C
VAL
18
13.128
−17.518
53.862
1.00
45.24
B


1724
O
VAL
18
13.072
−18.374
54.753
1.00
45.26
B


1725
N
VAL
19
12.978
−16.224
54.098
1.00
44.65
B


1726
CA
VAL
19
12.663
−15.758
55.435
1.00
44.13
B


1727
CB
VAL
19
13.926
−15.336
56.272
1.00
44.19
B


1728
CG1
VAL
19
14.885
−16.498
56.411
1.00
44.28
B


1729
CG2
VAL
19
14.611
−14.157
55.633
1.00
44.26
B


1730
C
VAL
19
11.764
−14.556
55.344
1.00
43.70
B


1731
O
VAL
19
11.831
−13.779
54.386
1.00
43.46
B


1732
N
THR
20
10.915
−14.421
56.350
1.00
43.23
B


1733
CA
THR
20
10.018
−13.294
56.448
1.00
43.00
B


1734
CB
THR
20
8.565
−13.743
56.490
1.00
43.00
B


1735
OG1
THR
20
8.244
−14.391
55.257
1.00
42.95
B


1736
CG2
THR
20
7.646
−12.540
56.691
1.00
42.85
B


1737
C
THR
20
10.368
−12.563
57.723
1.00
42.81
B


1738
O
THR
20
10.469
−13.157
58.791
1.00
42.81
B


1739
N
SER
21
10.560
−11.262
57.603
1.00
42.69
B


1740
CA
SER
21
10.938
−10.454
58.746
1.00
42.56
B


1741
CB
SER
21
11.586
−9.148
58.277
1.00
42.51
B


1742
OG
SER
21
11.680
−8.226
59.347
1.00
42.51
B


1743
C
SER
21
9.843
−10.101
59.728
1.00
42.46
B


1744
O
SER
21
8.838
−9.512
59.352
1.00
42.41
B


1745
N
PRO
22
10.022
−10.464
61.004
1.00
42.42
B


1746
CD
PRO
22
10.998
−11.374
61.631
1.00
42.39
B


1747
CA
PRO
22
8.974
−10.107
61.959
1.00
42.32
B


1748
CB
PRO
22
9.361
−10.890
63.208
1.00
42.38
B


1749
CG
PRO
22
10.839
−11.014
63.087
1.00
42.48
B


1750
C
PRO
22
9.035
−8.595
62.180
1.00
42.18
B


1751
O
PRO
22
8.143
−8.013
62.780
1.00
42.18
B


1752
N
VAL
23
10.100
−7.971
61.687
1.00
41.99
B


1753
CA
VAL
23
10.272
−6.532
61.845
1.00
41.75
B


1754
CB
VAL
23
11.781
−6.106
61.828
1.00
41.80
B


1755
CG1
VAL
23
11.887
−4.592
61.992
1.00
41.65
B


1756
CG2
VAL
23
12.534
−6.795
62.950
1.00
41.65
B


1757
C
VAL
23
9.561
−5.743
60.754
1.00
41.72
B


1758
O
VAL
23
8.772
−4.839
61.039
1.00
41.65
B


1759
N
THR
24
9.839
−6.076
59.500
1.00
41.57
B


1760
CA
THR
24
9.249
−5.340
58.386
1.00
41.68
B


1761
CB
THR
24
10.304
−4.993
57.325
1.00
41.54
B


1762
OG1
THR
24
10.806
−6.209
56.755
1.00
41.46
B


1763
CG2
THR
24
11.457
−4.206
57.946
1.00
41.58
B


1764
C
THR
24
8.135
−6.074
57.652
1.00
41.71
B


1765
O
THR
24
7.444
−5.478
56.825
1.00
41.85
B


1766
N
GLY
25
7.978
−7.358
57.934
1.00
41.81
B


1767
CA
GLY
25
6.951
−8.127
57.253
1.00
41.91
B


1768
C
GLY
25
7.378
−8.465
55.836
1.00
42.03
B


1769
O
GLY
25
6.656
−9.152
55.115
1.00
41.88
B


1770
N
LYS
26
8.552
−7.994
55.426
1.00
42.07
B


1771
CA
LYS
26
9.050
−8.268
54.081
1.00
42.16
B


1772
CB
LYS
26
10.130
−7.250
53.684
1.00
42.37
B


1773
CG
LYS
26
9.616
−5.819
53.685
1.00
42.59
B


1774
CD
LYS
26
10.656
−4.795
53.250
1.00
42.92
B


1775
CE
LYS
26
10.030
−3.404
53.240
1.00
43.24
B


1776
NZ
LYS
26
10.919
−2.330
52.711
1.00
43.72
B


1777
C
LYS
26
9.614
−9.674
54.008
1.00
42.21
B


1778
O
LYS
26
10.023
−10.238
55.022
1.00
42.21
B


1779
N
SER
27
9.623
−10.235
52.803
1.00
42.25
B


1780
CA
SER
27
10.142
−11.575
52.590
1.00
42.37
B


1781
CB
SER
27
9.086
−12.447
51.901
1.00
42.48
B


1782
OG
SER
27
7.938
−12.588
52.732
1.00
42.80
B


1783
C
SER
27
11.406
−11.512
51.748
1.00
42.40
B


1784
O
SER
27
11.514
−10.697
50.829
1.00
42.23
B


1785
N
TYR
28
12.358
−12.379
52.064
1.00
42.45
B


1786
CA
TYR
28
13.621
−12.408
51.351
1.00
42.71
B


1787
CB
TYR
28
14.770
−11.927
52.243
1.00
42.79
B


1788
CG
TYR
28
14.444
−10.811
53.208
1.00
42.90
B


1789
CD1
TYR
28
13.798
−11.073
54.416
1.00
43.14
B


1790
CE1
TYR
28
13.564
−10.052
55.344
1.00
43.31
B


1791
CD2
TYR
28
14.839
−9.500
52.943
1.00
42.89
B


1792
CE2
TYR
28
14.609
−8.477
53.855
1.00
42.94
B


1793
CZ
TYR
28
13.977
−8.756
55.054
1.00
43.14
B


1794
OH
TYR
28
13.790
−7.753
55.976
1.00
43.15
B


1795
C
TYR
28
13.957
−13.815
50.933
1.00
42.80
B


1796
O
TYR
28
13.404
−14.783
51.455
1.00
42.85
B


1797
N
PHE
29
14.883
−13.924
49.994
1.00
42.97
B


1798
CA
PHE
29
15.349
−15.224
49.551
1.00
43.23
B


1799
CB
PHE
29
15.030
−15.442
48.070
1.00
42.93
B


1800
CG
PHE
29
15.706
−14.477
47.153
1.00
42.69
B


1801
CD1
PHE
29
16.988
−14.731
46.678
1.00
42.62
B


1802
CD2
PHE
29
15.056
−13.313
46.750
1.00
42.65
B


1803
CE1
PHE
29
17.615
−13.840
45.814
1.00
42.52
B


1804
CE2
PHE
29
15.678
−12.415
45.886
1.00
42.52
B


1805
CZ
PHE
29
16.959
−12.684
45.416
1.00
42.46
B


1806
C
PHE
29
16.848
−15.224
49.810
1.00
43.57
B


1807
O
PHE
29
17.490
−14.175
49.766
1.00
43.42
B


1808
N
LEU
30
17.396
−16.394
50.106
1.00
44.16
B


1809
CA
LEU
30
18.815
−16.522
50.390
1.00
44.89
B


1810
CB
LEU
30
19.104
−17.902
50.977
1.00
44.66
B


1811
CG
LEU
30
20.547
−18.188
51.383
1.00
44.61
B


1812
CD1
LEU
30
20.981
−17.233
52.491
1.00
44.49
B


1813
CD2
LEU
30
20.650
−19.638
51.855
1.00
44.61
B


1814
C
LEU
30
19.673
−16.302
49.149
1.00
45.58
B


1815
O
LEU
30
19.652
−17.102
48.212
1.00
45.50
B


1816
N
GLY
31
20.428
−15.209
49.150
1.00
46.38
B


1817
CA
GLY
31
21.290
−14.909
48.022
1.00
47.43
B


1818
C
GLY
31
22.575
−15.705
48.108
1.00
48.27
B


1819
O
GLY
31
23.185
−16.041
47.091
1.00
48.24
B


1820
N
GLY
32
22.988
−16.013
49.333
1.00
49.11
B


1821
CA
GLY
32
24.204
−16.777
49.533
1.00
50.38
B


1822
C
GLY
32
24.594
−16.853
50.994
1.00
51.37
B


1823
O
GLY
32
24.123
−16.066
51.813
1.00
51.30
B


1824
N
ALA
33
25.449
−17.809
51.326
1.00
52.42
B


1825
CA
ALA
33
25.904
−17.972
52.698
1.00
53.70
B


1826
CB
ALA
33
25.011
−18.956
53.440
1.00
53.52
B


1827
C
ALA
33
27.332
−18.481
52.671
1.00
54.67
B


1828
O
ALA
33
27.717
−19.217
51.767
1.00
54.78
B


1829
N
GLY
34
28.119
−18.081
53.658
1.00
55.80
B


1830
CA
GLY
34
29.496
−18.525
53.712
1.00
57.28
B


1831
C
GLY
34
30.030
−18.472
55.124
1.00
58.38
B


1832
O
GLY
34
29.268
−18.376
56.085
1.00
58.39
B


1833
N
GLU
35
31.348
−18.537
55.250
1.00
59.51
B


1834
CA
GLU
35
31.990
−18.491
56.552
1.00
60.71
B


1835
CB
GLU
35
32.602
−19.853
56.878
1.00
60.83
B


1836
CG
GLU
35
33.477
−20.406
55.770
1.00
61.20
B


1837
CD
GLU
35
33.783
−21.883
55.949
1.00
61.39
B


1838
OE1
GLU
35
33.051
−22.562
56.704
1.00
61.47
B


1839
OE2
GLU
35
34.746
−22.369
55.319
1.00
61.58
B


1840
C
GLU
35
33.064
−17.415
56.576
1.00
61.50
B


1841
O
GLU
35
33.589
−17.021
55.533
1.00
61.60
B


1842
N
ARG
36
33.362
−16.932
57.774
1.00
62.40
B


1843
CA
ARG
36
34.377
−15.913
57.984
1.00
63.41
B


1844
CB
ARG
36
33.736
−14.531
58.136
1.00
63.24
B


1845
CG
ARG
36
33.101
−13.984
56.863
1.00
63.15
B


1846
CD
ARG
36
34.152
−13.434
55.898
1.00
63.03
B


1847
NE
ARG
36
33.553
−12.696
54.787
1.00
62.88
B


1848
CZ
ARG
36
32.857
−13.251
53.804
1.00
62.80
B


1849
NH1
ARG
36
32.666
−14.559
53.785
1.00
62.66
B


1850
NH2
ARG
36
32.357
−12.491
52.838
1.00
62.73
B


1851
C
ARG
36
35.102
−16.282
59.267
1.00
64.23
B


1852
O
ARG
36
34.516
−16.898
60.160
1.00
64.34
B


1853
N
GLY
37
36.372
−15.916
59.367
1.00
65.15
B


1854
CA
GLY
37
37.123
−16.241
60.564
1.00
66.33
B


1855
C
GLY
37
38.520
−15.667
60.540
1.00
67.19
B


1856
O
GLY
37
38.719
−14.513
60.155
1.00
67.22
B


1857
N
LEU
38
39.489
−16.473
60.961
1.00
68.07
B


1858
CA
LEU
38
40.879
−16.044
60.985
1.00
69.00
B


1859
CB
LEU
38
41.364
−15.882
62.429
1.00
69.01
B


1860
CG
LEU
38
40.616
−14.884
63.320
1.00
69.15
B


1861
CD1
LEU
38
41.184
−14.969
64.719
1.00
69.20
B


1862
CD2
LEU
38
40.743
−13.466
62.797
1.00
69.12
B


1863
C
LEU
38
41.768
−17.052
60.267
1.00
69.61
B


1864
O
LEU
38
41.450
−17.525
59.172
1.00
69.83
B


1865
N
THR
39
42.885
−17.363
60.908
1.00
70.30
B


1866
CA
THR
39
43.903
−18.295
60.432
1.00
70.92
B


1867
CB
THR
39
44.605
−17.816
59.125
1.00
71.00
B


1868
OG1
THR
39
44.875
−16.413
59.212
1.00
71.11
B


1869
CG2
THR
39
43.750
−18.112
57.893
1.00
71.08
B


1870
C
THR
39
44.933
−18.276
61.550
1.00
71.28
B


1871
O
THR
39
45.518
−17.230
61.849
1.00
71.39
B


1872
N
ILE
40
45.138
−19.414
62.191
1.00
71.65
B


1873
CA
ILE
40
46.104
−19.475
63.272
1.00
71.98
B


1874
CB
ILE
40
45.397
−19.519
64.649
1.00
72.07
B


1875
CG2
ILE
40
46.437
−19.577
65.768
1.00
72.13
B


1876
CG1
ILE
40
44.484
−18.293
64.789
1.00
72.16
B


1877
CD1
ILE
40
43.676
−18.247
66.072
1.00
72.19
B


1878
C
ILE
40
46.996
−20.695
63.113
1.00
72.13
B


1879
O
ILE
40
46.515
−21.829
63.098
1.00
72.14
B


1880
N
GLU
41
48.296
−20.445
62.990
1.00
72.27
B


1881
CA
GLU
41
49.280
−21.508
62.829
1.00
72.37
B


1882
CB
GLU
41
49.115
−22.567
63.924
1.00
72.66
B


1883
CG
GLU
41
49.074
−22.029
65.342
1.00
73.09
B


1884
CD
GLU
41
48.860
−23.131
66.370
1.00
73.44
B


1885
OE1
GLU
41
48.404
−24.239
65.982
1.00
73.61
B


1886
OE2
GLU
41
49.131
−22.891
67.573
1.00
73.57
B


1887
C
GLU
41
49.104
−22.154
61.461
1.00
72.24
B


1888
O
GLU
41
49.422
−23.333
61.284
1.00
72.33
B


1889
N
GLY
42
48.573
−21.388
60.509
1.00
72.00
B


1890
CA
GLY
42
48.378
−21.897
59.163
1.00
71.65
B


1891
C
GLY
42
47.039
−22.578
58.912
1.00
71.39
B


1892
O
GLY
42
46.765
−23.036
57.800
1.00
71.45
B


1893
N
ASN
43
46.198
−22.651
59.940
1.00
71.01
B


1894
CA
ASN
43
44.884
−23.285
59.808
1.00
70.49
B


1895
CB
ASN
43
44.699
−24.372
60.883
1.00
70.75
B


1896
CG
ASN
43
45.721
−24.277
62.002
1.00
70.91
B


1897
OD1
ASN
43
45.379
−24.076
63.167
1.00
71.10
B


1898
ND2
ASN
43
46.993
−24.420
61.651
1.00
70.99
B


1899
C
ASN
43
43.755
−22.266
59.908
1.00
69.94
B


1900
O
ASN
43
43.570
−21.639
60.955
1.00
69.92
B


1901
N
PHE
44
43.009
−22.091
58.819
1.00
69.17
B


1902
CA
PHE
44
41.893
−21.147
58.803
1.00
68.34
B


1903
CB
PHE
44
41.182
−21.163
57.449
1.00
68.55
B


1904
CG
PHE
44
40.096
−20.129
57.331
1.00
68.73
B


1905
CD1
PHE
44
40.406
−18.811
57.018
1.00
68.84
B


1906
CD2
PHE
44
38.767
−20.457
57.583
1.00
68.82
B


1907
CE1
PHE
44
39.410
−17.832
56.961
1.00
68.92
B


1908
CE2
PHE
44
37.766
−19.482
57.527
1.00
68.87
B


1909
CZ
PHE
44
38.091
−18.170
57.218
1.00
68.89
B


1910
C
PHE
44
40.886
−21.537
59.878
1.00
67.59
B


1911
O
PHE
44
40.255
−22.590
59.785
1.00
67.65
B


1912
N
ILE
45
40.721
−20.692
60.891
1.00
66.57
B


1913
CA
ILE
45
39.779
−21.003
61.958
1.00
65.46
B


1914
CB
ILE
45
40.346
−20.598
63.336
1.00
65.65
B


1915
CG2
ILE
45
39.326
−20.907
64.432
1.00
65.67
B


1916
CG1
ILE
45
41.658
−21.348
63.592
1.00
65.76
B


1917
CD1
ILE
45
42.299
−21.037
64.924
1.00
65.89
B


1918
C
ILE
45
38.444
−20.306
61.731
1.00
64.50
B


1919
O
ILE
45
38.384
−19.080
61.627
1.00
64.36
B


1920
N
LYS
46
37.383
−21.098
61.644
1.00
63.35
B


1921
CA
LYS
46
36.040
−20.569
61.422
1.00
62.15
B


1922
CB
LYS
46
35.113
−21.672
60.915
1.00
62.34
B


1923
CG
LYS
46
35.632
−22.408
59.702
1.00
62.53
B


1924
CD
LYS
46
34.607
−23.414
59.218
1.00
62.79
B


1925
CE
LYS
46
35.092
−24.169
57.994
1.00
62.95
B


1926
NZ
LYS
46
33.999
−24.987
57.410
1.00
63.14
B


1927
C
LYS
46
35.451
−19.992
62.702
1.00
61.12
B


1928
O
LYS
46
35.401
−20.669
63.727
1.00
61.07
B


1929
N
PHE
47
34.997
−18.747
62.635
1.00
59.85
B


1930
CA
PHE
47
34.398
−18.101
63.793
1.00
58.58
B


1931
CB
PHE
47
35.069
−16.763
64.080
1.00
58.88
B


1932
CG
PHE
47
36.336
−16.882
64.871
1.00
59.15
B


1933
CD1
PHE
47
37.512
−17.303
64.263
1.00
59.24
B


1934
CD2
PHE
47
36.352
−16.579
66.230
1.00
59.25
B


1935
CE1
PHE
47
38.691
−17.420
64.996
1.00
59.37
B


1936
CE2
PHE
47
37.530
−16.694
66.976
1.00
59.38
B


1937
CZ
PHE
47
38.702
−17.115
66.354
1.00
59.40
B


1938
C
PHE
47
32.909
−17.860
63.626
1.00
57.53
B


1939
O
PHE
47
32.149
−17.967
64.585
1.00
57.43
B


1940
N
THR
48
32.493
−17.526
62.411
1.00
56.23
B


1941
CA
THR
48
31.094
−17.242
62.153
1.00
54.89
B


1942
CB
THR
48
30.805
−15.740
62.306
1.00
54.93
B


1943
OG1
THR
48
31.464
−15.023
61.255
1.00
54.77
B


1944
CG2
THR
48
31.316
−15.221
63.639
1.00
54.93
B


1945
C
THR
48
30.647
−17.628
60.753
1.00
53.91
B


1946
O
THR
48
31.458
−17.826
59.848
1.00
53.77
B


1947
N
ALA
49
29.337
−17.736
60.594
1.00
52.68
B


1948
CA
ALA
49
28.736
−18.034
59.309
1.00
51.46
B


1949
CB
ALA
49
27.720
−19.153
59.445
1.00
51.35
B


1950
C
ALA
49
28.044
−16.722
58.953
1.00
50.62
B


1951
O
ALA
49
27.607
−15.989
59.844
1.00
50.37
B


1952
N
ILE
50
27.969
−16.412
57.667
1.00
49.66
B


1953
CA
ILE
50
27.323
−15.186
57.238
1.00
48.82
B


1954
CB
ILE
50
28.373
−14.144
56.789
1.00
49.03
B


1955
CG2
ILE
50
28.981
−14.557
55.462
1.00
49.23
B


1956
CG1
ILE
50
27.737
−12.761
56.694
1.00
49.22
B


1957
CD1
ILE
50
27.425
−12.141
58.065
1.00
49.28
B


1958
C
ILE
50
26.387
−15.529
56.086
1.00
48.02
B


1959
O
ILE
50
26.752
−16.270
55.179
1.00
48.00
B


1960
N
GLY
51
25.164
−15.023
56.154
1.00
47.11
B


1961
CA
GLY
51
24.206
−15.278
55.096
1.00
45.93
B


1962
C
GLY
51
23.618
−13.957
54.652
1.00
45.12
B


1963
O
GLY
51
23.386
−13.062
55.472
1.00
45.09
B


1964
N
VAL
52
23.388
−13.819
53.354
1.00
44.10
B


1965
CA
VAL
52
22.827
−12.590
52.826
1.00
43.04
B


1966
CB
VAL
52
23.757
−11.940
51.777
1.00
43.10
B


1967
CG1
VAL
52
23.093
−10.694
51.196
1.00
42.81
B


1968
CG2
VAL
52
25.092
−11.592
52.410
1.00
42.98
B


1969
C
VAL
52
21.489
−12.863
52.173
1.00
42.37
B


1970
O
VAL
52
21.397
−13.662
51.241
1.00
42.33
B


1971
N
TYR
53
20.456
−12.202
52.678
1.00
41.49
B


1972
CA
TYR
53
19.118
−12.339
52.134
1.00
40.80
B


1973
CB
TYR
53
18.089
−12.557
53.247
1.00
40.46
B


1974
CG
TYR
53
18.154
−13.924
53.872
1.00
40.32
B


1975
CD1
TYR
53
19.024
−14.190
54.929
1.00
40.13
B


1976
CE1
TYR
53
19.104
−15.465
55.492
1.00
40.11
B


1977
CD2
TYR
53
17.361
−14.969
53.388
1.00
40.22
B


1978
CE2
TYR
53
17.435
−16.249
53.946
1.00
40.15
B


1979
CZ
TYR
53
18.306
−26.488
54.992
1.00
40.03
B


1980
OH
TYR
53
18.397
−17.752
55.521
1.00
39.76
B


1981
C
TYR
53
18.746
−11.076
51.369
1.00
40.48
B


1982
O
TYR
53
19.054
−9.967
51.807
1.00
40.28
B


1983
N
LEU
54
18.080
−11.254
50.233
1.00
40.12
B


1984
CA
LEU
54
17.634
−10.140
49.409
1.00
39.95
B


1985
CB
LEU
54
18.195
−10.271
47.983
1.00
39.98
B


1986
CG
LEU
54
19.728
−10.346
47.857
1.00
39.84
B


1987
CD1
LEU
54
20.131
−10.370
46.390
1.00
40.00
B


1988
CD2
LEU
54
20.353
−9.146
48.568
1.00
39.77
B


1989
C
LEU
54
16.108
−10.168
49.373
1.00
39.95
B


1990
O
LEU
54
15.512
−11.231
49.197
1.00
40.06
B


1991
N
GLU
55
15.473
−9.015
49.554
1.00
39.84
B


1992
CA
GLU
55
14.017
−8.954
49.520
1.00
39.83
B


1993
CB
GLU
55
13.540
−7.505
49.619
1.00
39.82
B


1994
CG
GLU
55
12.101
−7.355
50.074
1.00
39.79
B


1995
CD
GLU
55
11.643
−5.912
50.094
1.00
40.01
B


1996
OE1
GLU
55
12.508
−5.004
50.125
1.00
39.92
B


1997
OE2
GLU
55
10.414
−5.683
50.089
1.00
40.17
B


1998
C
GLU
55
13.558
−9.563
48.194
1.00
39.88
B


1999
O
GLU
55
14.249
−9.448
47.178
1.00
39.68
B


2000
N
ASP
56
12.397
−10.211
48.200
1.00
39.86
B


2001
CA
ASP
56
11.899
−10.835
46.982
1.00
39.97
B


2002
CB
ASP
56
10.522
−11.481
47.227
1.00
40.55
B


2003
CG
ASP
56
9.551
−10.560
47.954
1.00
41.05
B


2004
OD1
ASP
56
9.792
−9.332
48.011
1.00
41.44
B


2005
OD2
ASP
56
8.526
−11.071
48.467
1.00
41.60
B


2006
C
ASP
56
11.837
−9.887
45.781
1.00
39.66
B


2007
O
ASP
56
12.165
−10.286
44.665
1.00
39.49
B


2008
N
ILE
57
11.442
−8.636
46.002
1.00
39.41
B


2009
CA
ILE
57
11.359
−7.678
44.900
1.00
39.38
B


2010
CB
ILE
57
10.753
−6.333
45.350
1.00
39.45
B


2011
CG2
ILE
57
9.325
−6.548
45.835
1.00
39.63
B


2012
CG1
ILE
57
11.623
−5.697
46.441
1.00
39.32
B


2013
CD1
ILE
57
11.202
−4.280
46.795
1.00
39.16
B


2014
C
ILE
57
12.702
−7.379
44.228
1.00
39.28
B


2015
O
ILE
57
12.751
−6.685
43.213
1.00
39.13
B


2016
N
ALA
58
13.792
−7.887
44.793
1.00
39.20
B


2017
CA
ALA
58
15.101
−7.646
44.201
1.00
39.17
B


2018
CB
ALA
58
16.196
−8.157
45.114
1.00
38.96
B


2019
C
ALA
58
15.202
−8.314
42.828
1.00
39.20
B


2020
O
ALA
58
15.903
−7.824
41.945
1.00
39.03
B


2021
N
VAL
59
14.503
−9.431
42.646
1.00
39.35
B


2022
CA
VAL
59
14.550
−10.130
41.360
1.00
39.57
B


2023
CB
VAL
59
13.663
−11.400
41.368
1.00
39.70
B


2024
CG1
VAL
59
13.666
−12.059
39.979
1.00
39.75
B


2025
CG2
VAL
59
14.182
−12.380
42.413
1.00
39.86
B


2026
C
VAL
59
14.107
−9.207
40.225
1.00
39.58
B


2027
O
VAL
59
14.806
−9.072
39.216
1.00
39.68
B


2028
N
ALA
60
12.960
−8.555
40.395
1.00
39.62
B


2029
CA
ALA
60
12.453
−7.651
39.368
1.00
39.75
B


2030
CB
ALA
60
11.058
−7.146
39.745
1.00
39.60
B


2031
C
ALA
60
13.400
−6.470
39.168
1.00
39.86
B


2032
O
ALA
60
13.582
−5.992
38.049
1.00
39.88
B


2033
N
SER
61
14.010
−6.007
40.256
1.00
39.96
B


2034
CA
SER
61
14.930
−4.883
40.182
1.00
40.10
B


2035
CB
SER
61
15.312
−4.435
41.599
1.00
40.24
B


2036
OG
SER
61
16.106
−3.268
41.565
1.00
40.32
B


2037
C
SER
61
16.190
−5.238
39.387
1.00
40.20
B


2038
O
SER
61
16.623
−4.491
38.511
1.00
39.93
B


2039
N
LEU
62
16.767
−6.396
39.681
1.00
40.48
B


2040
CA
LEU
62
17.984
−6.835
39.008
1.00
40.79
B


2041
CB
LEU
62
18.678
−7.889
39.873
1.00
40.83
B


2042
CG
LEU
62
19.092
−7.449
41.281
1.00
40.75
B


2043
CD1
LEU
62
19.447
−8.677
42.118
1.00
40.77
B


2044
CD2
LEU
62
20.276
−6.481
41.187
1.00
40.65
B


2045
C
LEU
62
17.770
−7.403
37.597
1.00
41.21
B


2046
O
LEU
62
18.699
−7.451
36.789
1.00
40.91
B


2047
N
ALA
63
16.545
−7.829
37.306
1.00
41.76
B


2048
CA
ALA
63
16.226
−8.428
36.014
1.00
42.24
B


2049
CB
ALA
63
14.758
−8.854
35.987
1.00
42.07
B


2050
C
ALA
63
16.526
−7.511
34.833
1.00
42.72
B


2051
O
ALA
63
17.105
−7.942
33.838
1.00
42.73
B


2052
N
ALA
64
16.134
−6.247
34.956
1.00
43.25
B


2053
CA
ALA
64
16.348
−5.258
33.900
1.00
43.82
B


2054
CB
ALA
64
16.090
−3.855
34.457
1.00
44.09
B


2055
C
ALA
64
17.746
−5.323
33.289
1.00
43.99
B


2056
O
ALA
64
17.905
−5.430
32.071
1.00
44.39
B


2057
N
LYS
65
18.758
−5.296
34.143
1.00
43.95
B


2058
CA
LYS
65
20.142
−5.309
33.691
1.00
44.01
B


2059
CB
LYS
65
20.988
−4.413
34.604
1.00
43.63
B


2060
CG
LYS
65
20.598
−2.951
34.605
1.00
43.43
B


2061
CD
LYS
65
21.298
−2.185
35.727
1.00
43.05
B


2062
CE
LYS
65
22.818
−2.233
35.597
1.00
42.89
B


2063
NZ
LYS
65
23.513
−1.392
36.619
1.00
42.53
B


2064
C
LYS
65
20.841
−6.659
33.620
1.00
44.17
B


2065
O
LYS
65
21.731
−6.857
32.789
1.00
44.16
B


2066
N
TRP
66
20.438
−7.590
34.475
1.00
44.43
B


2067
CA
TRP
66
21.143
−8.856
34.560
1.00
44.80
B


2068
CB
TRP
66
21.572
−9.045
36.017
1.00
44.39
B


2069
CG
TRP
66
22.255
−7.805
36.513
1.00
44.15
B


2070
CD2
TRP
66
23.492
−7.269
36.030
1.00
43.98
B


2071
CE2
TRP
66
23.682
−6.018
36.662
1.00
43.97
B


2072
CE3
TRP
66
24.449
−7.716
35.108
1.00
43.86
B


2073
CD1
TRP
66
21.767
−6.893
37.408
1.00
44.00
B


2074
NE1
TRP
66
22.619
−5.815
37.502
1.00
43.86
B


2075
CZ2
TRP
66
24.799
−5.215
36.411
1.00
43.93
B


2076
CZ3
TRP
66
25.559
−6.916
34.857
1.00
43.93
B


2077
CH2
TRP
66
25.720
−5.676
35.504
1.00
43.94
B


2078
C
TRP
66
20.528
−10.133
34.022
1.00
45.32
B


2079
O
TRP
66
21.208
−11.157
33.954
1.00
45.20
B


2080
N
LYS
67
19.261
−10.091
33.634
1.00
46.10
B


2081
CA
LYS
67
18.626
−11.292
33.100
1.00
46.95
B


2082
CB
LYS
67
17.176
−10.997
32.703
1.00
47.33
B


2083
CG
LYS
67
16.432
−12.217
32.173
1.00
48.05
B


2084
CD
LYS
67
14.974
−11.909
31.856
1.00
48.57
B


2085
CE
LYS
67
14.286
−13.147
31.276
1.00
48.96
B


2086
NZ
LYS
67
12.860
−12.897
30.895
1.00
49.49
B


2087
C
LYS
67
19.410
−11.802
31.888
1.00
47.21
B


2088
O
LYS
67
19.902
−11.013
31.083
1.00
47.25
B


2089
N
GLY
68
19.555
−13.120
31.773
1.00
47.53
B


2090
CA
GLY
68
20.264
−13.678
30.634
1.00
47.90
B


2091
C
GLY
68
21.770
−13.809
30.773
1.00
48.24
B


2092
O
GLY
68
22.407
−14.493
29.970
1.00
48.39
B


2093
N
LYS
69
22.346
−13.159
31.780
1.00
48.41
B


2094
CA
LYS
69
23.785
−13.222
32.014
1.00
48.60
B


2095
CB
LYS
69
24.231
−12.018
32.841
1.00
48.88
B


2096
CG
LYS
69
24.030
−10.681
32.143
1.00
49.27
B


2097
CD
LYS
69
25.263
−10.277
31.361
1.00
49.77
B


2098
CE
LYS
69
24.912
−9.242
30.306
1.00
50.02
B


2099
NZ
LYS
69
23.867
−8.311
30.829
1.00
50.38
B


2100
C
LYS
69
24.135
−14.510
32.754
1.00
48.62
B


2101
O
LYS
69
23.409
−14.937
33.649
1.00
48.58
B


2102
N
SER
70
25.247
−15.127
32.383
1.00
48.55
B


2103
CA
SER
70
25.667
−16.358
33.031
1.00
48.64
B


2104
CB
SER
70
26.741
−17.053
32.200
1.00
48.52
B


2105
OG
SER
70
27.937
−16.293
32.193
1.00
48.58
B


2106
C
SER
70
26.230
−16.045
34.411
1.00
48.68
B


2107
O
SER
70
26.568
−14.897
34.709
1.00
48.64
B


2108
N
SER
71
26.331
−17.070
35.248
1.00
48.70
B


2109
CA
SER
71
26.870
−16.891
36.586
1.00
48.84
B


2110
CB
SER
71
26.760
−18.197
37.382
1.00
48.74
B


2111
OG
SER
71
25.400
−18.558
37.570
1.00
48.45
B


2112
C
SER
71
28.328
−16.459
36.490
1.00
49.02
B


2113
O
SER
71
28.801
−15.635
37.274
1.00
48.97
B


2114
N
GLU
72
29.031
−17.015
35.510
1.00
49.24
B


2115
CA
GLU
72
30.436
−16.702
35.299
1.00
49.50
B


2116
CB
GLU
72
31.000
−17.565
34.167
1.00
50.10
B


2117
CG
GLU
72
30.731
−19.074
34.306
1.00
51.11
B


2118
CD
GLU
72
29.272
−19.459
34.048
1.00
51.70
B


2119
OE1
GLU
72
28.670
−18.953
33.078
1.00
52.50
B


2120
OE2
GLU
72
28.721
−20.286
34.803
1.00
52.20
B


2121
C
GLU
72
30.600
−15.224
34.948
1.00
49.31
B


2122
O
GLU
72
31.546
−14.567
35.397
1.00
49.38
B


2123
N
GLU
73
29.682
−14.701
34.139
1.00
49.02
B


2124
CA
GLU
73
29.761
−13.298
33.749
1.00
48.82
B


2125
CB
GLU
73
28.801
−12.983
32.597
1.00
49.24
B


2126
CG
GLU
73
29.011
−11.570
32.047
1.00
50.00
B


2127
CD
GLU
73
28.126
−11.233
30.861
1.00
50.48
B


2128
OE1
GLU
73
27.590
−12.169
30.214
1.00
50.83
B


2129
OE2
GLU
73
27.983
−10.021
30.572
1.00
50.72
B


2130
C
GLU
73
29.439
−12.401
34.937
1.00
48.32
B


2131
O
GLU
73
30.153
−11.434
35.212
1.00
48.19
B


2132
N
LEU
74
28.360
−12.723
35.640
1.00
47.81
B


2133
CA
LEU
74
27.957
−11.938
36.800
1.00
47.37
B


2134
CB
LEU
74
26.719
−12.559
37.455
1.00
47.26
B


2135
CG
LEU
74
25.433
−12.529
36.630
1.00
47.22
B


2136
CD1
LEU
74
24.321
−13.215
37.400
1.00
47.15
B


2137
CD2
LEU
74
25.057
−11.085
36.320
1.00
47.11
B


2138
C
LEU
74
29.078
−11.837
37.827
1.00
47.06
B


2139
O
LEU
74
29.380
−10.754
38.324
1.00
46.87
B


2140
N
LEU
75
29.705
−12.966
38.137
1.00
46.79
B


2141
CA
LEU
75
30.771
−12.980
39.131
1.00
46.59
B


2142
CB
LEU
75
31.366
−14.385
39.251
1.00
46.57
B


2143
CG
LEU
75
32.465
−14.537
40.306
1.00
46.63
B


2144
CD1
LEU
75
31.920
−14.130
41.670
1.00
46.55
B


2145
CD2
LEU
75
32.967
−15.977
40.330
1.00
46.55
B


2146
C
LEU
75
31.881
−11.969
38.849
1.00
46.50
B


2147
O
LEU
75
32.430
−11.361
39.772
1.00
46.44
B


2148
N
GLU
76
32.210
−11.779
37.574
1.00
46.41
B


2149
CA
GLU
76
33.270
−10.844
37.205
1.00
46.29
B


2150
CB
GLU
76
34.048
−11.381
35.995
1.00
47.04
B


2151
CG
GLU
76
33.176
−12.071
34.936
1.00
48.20
B


2152
CD
GLU
76
33.934
−12.441
33.654
1.00
48.84
B


2153
OE1
GLU
76
35.182
−12.559
33.696
1.00
49.33
B


2154
OE2
GLU
76
33.272
−12.626
32.600
1.00
49.21
B


2155
C
GLU
76
32.775
−9.420
36.912
1.00
45.70
B


2156
O
GLU
76
33.538
−8.580
36.438
1.00
45.76
B


2157
N
THR
77
31.507
−9.145
37.199
1.00
44.89
B


2158
CA
THR
77
30.948
−7.815
36.945
1.00
44.08
B


2159
CB
THR
77
29.638
−7.920
36.156
1.00
44.08
B


2160
OG1
THR
77
29.855
−8.706
34.982
1.00
44.04
B


2161
CG2
THR
77
29.148
−6.536
35.760
1.00
44.00
B


2162
C
THR
77
30.682
−7.043
38.230
1.00
43.44
B


2163
O
THR
77
29.687
−7.281
38.913
1.00
43.38
B


2164
N
LEU
78
31.573
−6.115
38.556
1.00
42.63
B


2165
CA
LEU
78
31.425
−5.327
39.768
1.00
41.89
B


2166
CB
LEU
78
32.560
−4.311
39.903
1.00
41.84
B


2167
CG
LEU
78
33.971
−4.843
40.166
1.00
41.77
B


2168
CD1
LEU
78
34.936
−3.655
40.283
1.00
41.72
B


2169
CD2
LEU
78
33.984
−5.680
41.445
1.00
41.60
B


2170
C
LEU
78
30.101
−4.591
39.796
1.00
41.50
B


2171
O
LEU
78
29.509
−4.410
40.863
1.00
41.40
B


2172
N
ASP
79
29.641
−4.166
38.625
1.00
40.76
B


2173
CA
ASP
79
28.391
−3.436
38.537
1.00
40.19
B


2174
CB
ASP
79
28.133
−2.994
37.099
1.00
39.93
B


2175
CG
ASP
79
27.021
−1.973
37.005
1.00
39.73
B


2176
OD1
ASP
79
27.080
−.976
37.758
1.00
39.52
B


2177
OD2
ASP
79
26.094
−2.163
36.188
1.00
39.68
B


2178
C
ASP
79
27.222
−4.274
39.046
1.00
39.95
B


2179
O
ASP
79
26.277
−3.741
39.620
1.00
39.93
B


2180
N
PHE
80
27.290
−5.584
38.833
1.00
39.65
B


2181
CA
PHE
80
26.238
−6.479
39.298
1.00
39.49
B


2182
CB
PHE
80
26.535
−7.914
38.848
1.00
39.69
B


2183
CG
PHE
80
25.635
−8.947
39.471
1.00
39.88
B


2184
CD1
PHE
80
26.171
−9.978
40.232
1.00
39.84
B


2185
CD2
PHE
80
24.257
−8.893
39.290
1.00
39.82
B


2186
CE1
PHE
80
25.349
−10.944
40.804
1.00
39.98
B


2187
CE2
PHE
80
23.427
−9.854
39.857
1.00
39.99
B


2188
CZ
PHE
80
23.971
−10.882
40.615
1.00
39.89
B


2189
C
PHE
80
26.141
−6.419
40.822
1.00
39.23
B


2190
O
PHE
80
25.053
−6.280
41.388
1.00
39.29
B


2191
N
TYR
81
27.281
−6.513
41.494
1.00
38.84
B


2192
CA
TYR
81
27.268
−6.466
42.947
1.00
38.60
B


2193
CB
TYR
81
28.603
−6.960
43.500
1.00
38.64
B


2194
CG
TYR
81
28.822
−8.407
43.146
1.00
38.74
B


2195
CD1
TYR
81
29.522
−8.766
41.993
1.00
38.91
B


2196
CE1
TYR
81
29.624
−10.097
41.598
1.00
38.98
B


2197
CD2
TYR
81
28.236
−9.418
43.901
1.00
38.88
B


2198
CE2
TYR
81
28.330
−10.756
43.514
1.00
38.83
B


2199
CZ
TYR
81
29.026
−11.085
42.362
1.00
39.00
B


2200
OH
TYR
81
29.124
−12.401
41.968
1.00
39.16
B


2201
C
TYR
81
26.915
−5.090
43.491
1.00
38.39
B


2202
O
TYR
81
26.368
−4.983
44.582
1.00
38.23
B


2203
N
ARG
82
27.209
−4.036
42.735
1.00
38.27
B


2204
CA
ARG
82
26.855
−2.700
43.194
1.00
38.20
B


2205
CB
ARG
82
27.430
−1.615
42.277
1.00
38.62
B


2206
CG
ARG
82
28.948
−1.499
42.343
1.00
39.18
B


2207
CD
ARG
82
29.415
−.058
42.150
1.00
39.76
B


2208
NE
ARG
82
30.865
.032
42.296
1.00
40.24
B


2209
CZ
ARG
82
31.732
−.267
41.333
1.00
40.54
B


2210
NH1
ARG
82
31.294
−.660
40.142
1.00
40.61
B


2211
NH2
ARG
82
33.039
−.202
41.573
1.00
40.71
B


2212
C
ARG
82
25.335
−2.610
43.218
1.00
37.97
B


2213
O
ARG
82
24.762
−2.015
44.131
1.00
37.74
B


2214
N
ASP
83
24.683
−3.209
42.220
1.00
37.78
B


2215
CA
ASP
83
23.223
−3.189
42.175
1.00
37.70
B


2216
CB
ASP
83
22.690
−3.795
40.873
1.00
37.72
B


2217
CG
ASP
83
22.784
−2.835
39.699
1.00
37.89
B


2218
OD1
ASP
83
22.683
−1.604
39.917
1.00
38.14
B


2219
OD2
ASP
83
22.931
−3.306
38.556
1.00
37.83
B


2220
C
ASP
83
22.664
−3.960
43.366
1.00
37.51
B


2221
O
ASP
83
21.676
−3.559
43.958
1.00
37.53
B


2222
N
ILE
84
23.300
−5.066
43.725
1.00
37.44
B


2223
CA
ILE
84
22.825
−5.832
44.863
1.00
37.47
B


2224
CB
ILE
84
23.555
−7.184
44.978
1.00
37.40
B


2225
CG2
ILE
84
23.210
−7.854
46.304
1.00
37.28
B


2226
CG1
ILE
84
23.161
−8.074
43.795
1.00
37.35
B


2227
CD1
ILE
84
23.852
−9.419
43.769
1.00
37.46
B


2228
C
ILE
84
23.021
−5.035
46.150
1.00
37.47
B


2229
O
ILE
84
22.121
−4.953
46.976
1.00
37.59
B


2230
N
ILE
85
24.191
−4.428
46.311
1.00
37.43
B


2231
CA
ILE
85
24.481
−3.646
47.511
1.00
37.40
B


2232
CB
ILE
85
25.951
−3.156
47.512
1.00
37.47
B


2233
CG2
ILE
85
26.181
−2.201
48.678
1.00
37.29
B


2234
CG1
ILE
85
26.905
−4.352
47.594
1.00
37.36
B


2235
CD1
ILE
85
28.350
−3.996
47.266
1.00
37.35
B


2236
C
ILE
85
23.583
−2.418
47.684
1.00
37.50
B


2237
O
ILE
85
23.046
−2.178
48.770
1.00
37.33
B


2238
N
SER
86
23.420
−1.636
46.620
1.00
37.58
B


2239
CA
SER
86
22.611
−.426
46.719
1.00
37.89
B


2240
CB
SER
86
23.423
.769
46.218
1.00
38.16
B


2241
OG
SER
86
24.648
.859
46.930
1.00
38.50
B


2242
C
SER
86
21.277
−.494
45.982
1.00
37.85
B


2243
O
SER
86
20.663
.541
45.697
1.00
37.88
B


2244
N
GLY
87
20.833
−1.712
45.684
1.00
37.83
B


2245
CA
GLY
87
19.570
−1.901
44.993
1.00
37.79
B


2246
C
GLY
87
18.395
−1.401
45.814
1.00
37.71
B


2247
O
GLY
87
18.460
−1.388
47.042
1.00
37.67
B


2248
N
PRO
88
17.297
−.997
45.157
1.00
37.71
B


2249
CD
PRO
88
17.139
−1.053
43.692
1.00
37.67
B


2250
CA
PRO
88
16.080
−.479
45.791
1.00
37.83
B


2251
CB
PRO
88
15.341
.154
44.618
1.00
37.69
B


2252
CG
PRO
88
15.643
−.807
43.510
1.00
37.64
B


2253
C
PRO
88
15.244
−1.541
46.500
1.00
37.94
B


2254
O
PRO
88
14.056
−1.713
46.216
1.00
37.87
B


2255
N
PHE
89
15.870
−2.244
47.431
1.00
38.07
B


2256
CA
PHE
89
15.194
−3.290
48.180
1.00
38.25
B


2257
CB
PHE
89
15.063
−4.550
47.324
1.00
38.10
B


2258
CG
PHE
89
16.319
−4.907
46.578
1.00
38.02
B


2259
CD1
PHE
89
17.376
−5.543
47.225
1.00
37.76
B


2260
CD2
PHE
89
16.449
−4.589
45.220
1.00
37.82
B


2261
CE1
PHE
89
18.543
−5.858
46.533
1.00
37.81
B


2262
CE2
PHE
89
17.612
−4.899
44.521
1.00
37.79
B


2263
CZ
PHE
89
18.662
−5.536
45.180
1.00
37.99
B


2264
C
PHE
89
15.977
−3.590
49.441
1.00
38.46
B


2265
O
PHE
89
17.172
−3.309
49.529
1.00
38.42
B


2266
N
GLU
90
15.296
−4.172
50.413
1.00
38.66
B


2267
CA
GLU
90
15.917
−4.485
51.681
1.00
38.95
B


2268
CB
GLU
90
14.829
−4.737
52.723
1.00
39.19
B


2269
CG
GLU
90
15.307
−4.613
54.146
1.00
39.63
B


2270
CD
GLU
90
14.167
−4.448
55.112
1.00
39.63
B


2271
OE1
GLU
90
13.588
−5.471
55.534
1.00
39.90
B


2272
OE2
GLU
90
13.842
−3.291
55.441
1.00
39.82
B


2273
C
GLU
90
16.828
−5.692
51.591
1.00
39.01
B


2274
O
GLU
90
16.562
−6.632
50.836
1.00
39.01
B


2275
N
LYS
91
17.929
−5.646
52.333
1.00
38.99
B


2276
CA
LYS
91
18.838
−6.778
52.389
1.00
39.10
B


2277
CB
LYS
91
20.238
−6.431
51.873
1.00
39.04
B


2278
CG
LYS
91
20.282
−5.909
50.435
1.00
39.18
B


2279
CD
LYS
91
20.188
−4.385
50.407
1.00
39.16
B


2280
CE
LYS
91
20.294
−3.844
48.993
1.00
39.09
B


2281
NZ
LYS
91
20.328
−2.354
49.026
1.00
39.27
B


2282
C
LYS
91
18.892
−7.138
53.865
1.00
39.23
B


2283
O
LYS
91
18.820
−6.273
54.738
1.00
39.21
B


2284
N
LEU
92
18.986
−8.423
54.146
1.00
39.40
B


2285
CA
LEU
92
19.034
−8.876
55.522
1.00
39.58
B


2286
CB
LEU
92
17.745
−9.628
55.873
1.00
39.62
B


2287
CG
LEU
92
17.642
−10.163
57.300
1.00
39.75
B


2288
CD1
LEU
92
17.771
−9.027
58.303
1.00
39.86
B


2289
CD2
LEU
92
16.304
−10.867
57.465
1.00
39.95
B


2290
C
LEU
92
20.230
−9.790
55.639
1.00
39.65
B


2291
O
LEU
92
20.281
−10.847
55.013
1.00
39.55
B


2292
N
ILE
93
21.199
−9.370
56.439
1.00
39.89
B


2293
CA
ILE
93
22.412
−10.145
56.627
1.00
40.18
B


2294
CB
ILE
93
23.651
−9.237
56.540
1.00
40.36
B


2295
CG2
ILE
93
24.919
−10.075
56.614
1.00
40.39
B


2296
CG1
ILE
93
23.623
−8.436
55.237
1.00
40.47
B


2297
CD1
ILE
93
24.658
−7.332
55.209
1.00
41.03
B


2298
C
ILE
93
22.390
−10.806
57.996
1.00
40.37
B


2299
O
ILE
93
22.205
−10.136
59.011
1.00
40.20
B


2300
N
ARG
94
22.561
−12.122
58.030
1.00
40.70
B


2301
CA
ARG
94
22.578
−12.813
59.304
1.00
41.11
B


2302
CB
ARG
94
21.511
−13.905
59.378
1.00
41.34
B


2303
CG
ARG
94
21.391
−14.492
60.776
1.00
41.86
B


2304
CD
ARG
94
21.838
−15.945
60.864
1.00
42.47
B


2305
NE
ARG
94
20.839
−16.837
60.298
1.00
42.93
B


2306
CZ
ARG
94
20.929
−18.163
60.275
1.00
43.21
B


2307
NH1
ARG
94
21.980
−18.783
60.794
1.00
43.37
B


2308
NH2
ARG
94
19.954
−18.873
59.721
1.00
43.51
B


2309
C
ARG
94
23.930
−13.447
59.561
1.00
41.29
B


2310
O
ARG
94
24.403
−14.272
58.782
1.00
41.31
B


2311
N
GLY
95
24.559
−13.031
60.650
1.00
41.47
B


2312
CA
GLY
95
25.831
−13.608
61.021
1.00
41.78
B


2313
C
GLY
95
25.524
−14.484
62.217
1.00
42.11
B


2314
O
GLY
95
24.856
−14.038
63.148
1.00
41.97
B


2315
N
SER
96
25.968
−15.734
62.180
1.00
42.50
B


2316
CA
SER
96
25.753
−16.659
63.286
1.00
43.05
B


2317
CB
SER
96
24.888
−17.839
62.845
1.00
43.13
B


2318
OG
SER
96
23.562
−17.402
62.592
1.00
43.17
B


2319
C
SER
96
27.116
−17.147
63.742
1.00
43.42
B


2320
O
SER
96
27.966
−17.479
62.920
1.00
43.38
B


2321
N
LYS
97
27.326
−17.181
65.053
1.00
43.78
B


2322
CA
LYS
97
28.613
−17.601
65.594
1.00
44.33
B


2323
CB
LYS
97
28.782
−17.055
67.016
1.00
43.97
B


2324
CG
LYS
97
29.334
−15.636
67.063
1.00
43.61
B


2325
CD
LYS
97
28.487
−14.660
66.262
1.00
43.15
B


2326
CE
LYS
97
29.113
−13.271
66.287
1.00
42.99
B


2327
NZ
LYS
97
28.302
−12.273
65.553
1.00
42.83
B


2328
C
LYS
97
28.888
−19.094
65.599
1.00
44.89
B


2329
O
LYS
97
28.017
−19.907
65.920
1.00
45.01
B


2330
N
ILE
98
30.111
−19.446
65.218
1.00
45.54
B


2331
CA
ILE
98
30.547
−20.832
65.227
1.00
46.25
B


2332
CB
ILE
98
31.464
−21.135
64.031
1.00
46.31
B


2333
CG2
ILE
98
32.038
−22.541
64.159
1.00
46.33
B


2334
CG1
ILE
98
30.663
−20.998
62.732
1.00
46.41
B


2335
CD1
ILE
98
31.501
−21.063
61.460
1.00
46.56
B


2336
C
ILE
98
31.313
−20.958
66.546
1.00
46.69
B


2337
O
ILE
98
31.185
−21.955
67.258
1.00
46.81
B


2338
N
ARG
99
32.093
−19.928
66.862
1.00
47.22
B


2339
CA
ARG
99
32.839
−19.859
68.115
1.00
47.79
B


2340
CB
ARG
99
34.341
−19.653
67.876
1.00
48.66
B


2341
CG
ARG
99
35.039
−20.829
67.200
1.00
50.10
B


2342
CD
ARG
99
36.549
−20.590
67.108
1.00
51.29
B


2343
NE
ARG
99
37.194
−21.602
66.272
1.00
52.52
B


2344
CZ
ARG
99
37.376
−22.872
66.630
1.00
52.95
B


2345
NH1
ARG
99
36.967
−23.293
67.821
1.00
53.15
B


2346
NH2
ARG
99
37.957
−23.727
65.791
1.00
53.37
B


2347
C
ARG
99
32.288
−18.656
68.873
1.00
47.56
B


2348
O
ARG
99
31.841
−17.685
68.263
1.00
47.62
B


2349
N
GLU
100
32.320
−18.721
70.192
1.00
47.26
B


2350
CA
GLU
100
31.806
−17.635
71.011
1.00
46.82
B


2351
CB
GLU
100
31.807
−18.042
72.482
1.00
47.00
B


2352
CG
GLU
100
31.353
−16.941
73.415
1.00
47.32
B


2353
CD
GLU
100
31.344
−17.385
74.861
1.00
47.64
B


2354
OE1
GLU
100
32.328
−18.032
75.291
1.00
47.96
B


2355
OE2
GLU
100
30.361
−17.083
75.569
1.00
47.60
B


2356
C
GLU
100
32.613
−16.363
70.831
1.00
46.46
B


2357
O
GLU
100
33.840
−16.388
70.784
1.00
46.42
B


2358
N
LEU
101
31.911
−15.244
70.730
1.00
45.98
B


2359
CA
LEU
101
32.542
−13.945
70.558
1.00
45.58
B


2360
CB
LEU
101
32.636
−13.575
69.074
1.00
45.63
B


2361
CG
LEU
101
33.666
−14.276
68.178
1.00
45.72
B


2362
CD1
LEU
101
33.383
−13.944
66.717
1.00
45.66
B


2363
CD2
LEU
101
35.072
−13.833
68.564
1.00
45.72
B


2364
C
LEU
101
31.690
−12.913
71.262
1.00
45.28
B


2365
O
LEU
101
30.467
−12.924
71.133
1.00
45.19
B


2366
N
SER
102
32.328
−12.029
72.016
1.00
44.93
B


2367
CA
SER
102
31.591
−10.982
72.693
1.00
44.64
B


2368
CB
SER
102
32.470
−10.285
73.728
1.00
44.63
B


2369
OG
SER
102
33.484
−9.537
73.091
1.00
44.35
B


2370
C
SER
102
31.216
−9.995
71.598
1.00
44.52
B


2371
O
SER
102
31.745
−10.065
70.481
1.00
44.43
B


2372
N
GLY
103
30.309
−9.080
71.913
1.00
44.33
B


2373
CA
GLY
103
29.899
−8.098
70.931
1.00
44.23
B


2374
C
GLY
103
31.084
−7.273
70.461
1.00
44.21
B


2375
O
GLY
103
31.335
−7.179
69.257
1.00
43.93
B


2376
N
PRO
104
31.835
−6.657
71.391
1.00
44.23
B


2377
CD
PRO
104
31.601
−6.583
72.847
1.00
44.17
B


2378
CA
PRO
104
32.995
−5.845
71.003
1.00
44.29
B


2379
CB
PRO
104
33.556
−5.384
72.349
1.00
44.28
B


2380
CG
PRO
104
32.315
−5.298
73.219
1.00
44.22
B


2381
C
PRO
104
34.018
−6.640
70.184
1.00
44.41
B


2382
O
PRO
104
34.577
−6.132
69.204
1.00
44.34
B


2383
N
GLU
105
34.252
−7.888
70.583
1.00
44.57
B


2384
CA
GLU
105
35.211
−8.744
69.889
1.00
44.86
B


2385
CB
GLU
105
35.313
−10.107
70.589
1.00
45.23
B


2386
CG
GLU
105
36.169
−10.108
71.864
1.00
45.87
B


2387
CD
GLU
105
35.997
−11.374
72.700
1.00
46.21
B


2388
OE1
GLU
105
35.568
−12.412
72.141
1.00
46.29
B


2389
OE2
GLU
105
36.303
−11.328
73.919
1.00
46.63
B


2390
C
GLU
105
34.812
−8.948
68.431
1.00
44.86
B


2391
O
GLU
105
35.628
−8.784
67.521
1.00
44.82
B


2392
N
TYR
106
33.550
−9.306
68.214
1.00
44.74
B


2393
CA
TYR
106
33.044
−9.542
66.869
1.00
44.72
B


2394
CB
TYR
106
31.650
−10.170
66.924
1.00
44.32
B


2395
CG
TYR
106
30.890
−10.017
65.623
1.00
43.89
B


2396
CD1
TYR
106
31.158
−10.844
64.531
1.00
43.61
B


2397
CE1
TYR
106
30.499
−10.660
63.312
1.00
43.43
B


2398
CD2
TYR
106
29.943
−9.002
65.467
1.00
43.68
B


2399
CE2
TYR
106
29.284
−8.807
64.259
1.00
43.45
B


2400
CZ
TYR
106
29.565
−9.634
63.185
1.00
43.44
B


2401
OH
TYR
106
28.934
−9.412
61.981
1.00
43.19
B


2402
C
TYR
106
32.969
−8.265
66.044
1.00
44.85
B


2403
O
TYR
106
33.394
−8.239
64.895
1.00
44.94
B


2404
N
SER
107
32.428
−7.207
66.637
1.00
45.03
B


2405
CA
SER
107
32.253
−5.940
65.943
1.00
45.36
B


2406
CB
SER
107
31.366
−5.008
66.772
1.00
45.31
B


2407
OG
SER
107
32.028
−4.577
67.948
1.00
45.62
B


2408
C
SER
107
33.525
−5.192
65.554
1.00
45.54
B


2409
O
SER
107
33.516
−4.416
64.589
1.00
45.33
B


2410
N
ARG
108
34.614
−5.411
66.289
1.00
45.82
B


2411
CA
ARG
108
35.862
−4.707
65.998
1.00
46.19
B


2412
CB
ARG
108
37.010
−5.245
66.860
1.00
46.83
B


2413
CG
ARG
108
38.393
−4.627
66.567
1.00
47.97
B


2414
CD
ARG
108
39.447
−5.200
67.510
1.00
48.98
B


2415
NE
ARG
108
39.228
−6.632
67.687
1.00
50.09
B


2416
CZ
ARG
108
39.654
−7.573
66.848
1.00
50.65
B


2417
NH1
ARG
108
40.350
−7.241
65.764
1.00
50.78
B


2418
NH2
ARG
108
39.341
−8.851
67.074
1.00
51.14
B


2419
C
ARG
108
36.253
−4.758
64.522
1.00
46.00
B


2420
O
ARG
108
36.381
−3.721
63.881
1.00
45.86
B


2421
N
LYS
109
36.429
−5.957
63.982
1.00
45.88
B


2422
CA
LYS
109
36.824
−6.105
62.585
1.00
45.94
B


2423
CB
LYS
109
37.112
−7.574
62.269
1.00
46.46
B


2424
CG
LYS
109
37.774
−7.804
60.906
1.00
47.39
B


2425
CD
LYS
109
39.186
−7.201
60.841
1.00
48.10
B


2426
CE
LYS
109
39.836
−7.472
59.481
1.00
48.60
B


2427
NZ
LYS
109
41.268
−7.022
59.421
1.00
49.19
B


2428
C
LYS
109
35.760
−5.565
61.630
1.00
45.63
B


2429
O
LYS
109
36.079
−4.855
60.670
1.00
45.55
B


2430
N
VAL
110
34.497
−5.886
61.894
1.00
45.16
B


2431
CA
VAL
110
33.410
−5.414
61.051
1.00
44.73
B


2432
CB
VAL
110
32.028
−5.853
61.593
1.00
44.71
B


2433
CG1
VAL
110
30.907
−5.239
60.748
1.00
44.63
B


2434
CG2
VAL
110
31.929
−7.369
61.574
1.00
44.80
B


2435
C
VAL
110
33.432
−3.894
60.944
1.00
44.42
B


2436
O
VAL
110
33.358
−3.342
59.846
1.00
44.16
B


2437
N
MET
111
33.544
−3.219
62.081
1.00
44.16
B


2438
CA
MET
111
33.568
−1.766
62.091
1.00
44.20
B


2439
CB
MET
111
33.521
−1.241
63.526
1.00
44.23
B


2440
CG
MET
111
32.216
−1.562
64.241
1.00
44.37
B


2441
SD
MET
111
32.026
−.698
65.800
1.00
44.68
B


2442
CE
MET
111
33.149
−1.674
66.836
1.00
44.57
B


2443
C
MET
111
34.795
−1.223
61.365
1.00
44.19
B


2444
O
MET
111
34.722
−.204
60.676
1.00
44.09
B


2445
N
GLU
112
35.919
−1.913
61.523
1.00
44.23
B


2446
CA
GLU
112
37.159
−1.521
60.865
1.00
44.24
B


2447
CB
GLU
112
38.259
−2.551
61.153
1.00
44.74
B


2448
CG
GLU
112
39.006
−2.344
62.463
1.00
45.81
B


2449
CD
GLU
112
40.078
−3.403
62.683
1.00
46.41
B


2450
OE1
GLU
112
40.665
−3.867
61.674
1.00
47.00
B


2451
OE2
GLU
112
40.345
−3.760
63.857
1.00
46.99
B


2452
C
GLU
112
36.935
−1.453
59.359
1.00
43.86
B


2453
O
GLU
112
37.242
−.454
58.710
1.00
43.75
B


2454
N
ASN
113
36.394
−2.544
58.824
1.00
43.53
B


2455
CA
ASN
113
36.133
−2.674
57.400
1.00
43.29
B


2456
CB
ASN
113
35.655
−4.086
57.066
1.00
43.39
B


2457
CG
ASN
113
36.726
−5.126
57.312
1.00
43.65
B


2458
OD1
ASN
113
37.915
−4.822
57.266
1.00
43.96
B


2459
ND2
ASN
113
36.318
−6.355
57.557
1.00
43.65
B


2460
C
ASN
113
35.121
−1.656
56.927
1.00
43.03
B


2461
O
ASN
113
35.241
−1.141
55.821
1.00
43.01
B


2462
N
CYS
114
34.126
−1.361
57.755
1.00
42.64
B


2463
CA
CYS
114
33.133
−.374
57.364
1.00
42.52
B


2464
CB
CYS
114
31.963
−.336
58.359
1.00
42.13
B


2465
SG
CYS
114
30.837
−1.760
58.238
1.00
41.13
B


2466
C
CYS
114
33.771
1.005
57.283
1.00
42.70
B


2467
O
CYS
114
33.579
1.724
56.308
1.00
42.60
B


2468
N
VAL
115
34.526
1.372
58.313
1.00
42.95
B


2469
CA
VAL
115
35.173
2.676
58.337
1.00
43.35
B


2470
CB
VAL
115
35.923
2.917
59.668
1.00
43.35
B


2471
CG1
VAL
115
36.626
4.277
59.630
1.00
43.27
B


2472
CG2
VAL
115
34.945
2.870
60.836
1.00
43.13
B


2473
C
VAL
115
36.157
2.822
57.177
1.00
43.62
B


2474
O
VAL
115
36.255
3.893
56.577
1.00
43.60
B


2475
N
ALA
116
36.882
1.751
56.864
1.00
43.95
B


2476
CA
ALA
116
37.840
1.787
55.764
1.00
44.42
B


2477
CB
ALA
116
38.616
.471
55.688
1.00
44.43
B


2478
C
ALA
116
37.087
2.033
54.456
1.00
44.80
B


2479
O
ALA
116
37.505
2.844
53.627
1.00
44.82
B


2480
N
HIS
117
35.971
1.335
54.270
1.00
45.02
B


2481
CA
HIS
117
35.182
1.517
53.059
1.00
45.36
B


2482
CB
HIS
117
33.978
.577
53.042
1.00
45.33
B


2483
CG
HIS
117
32.889
1.029
52.119
1.00
45.35
B


2484
CD2
HIS
117
31.675
1.569
52.371
1.00
45.40
B


2485
ND1
HIS
117
33.023
1.008
50.746
1.00
45.38
B


2486
CE1
HIS
117
31.935
1.516
50.194
1.00
45.41
B


2487
NE2
HIS
117
31.100
1.865
51.156
1.00
45.50
B


2488
C
HIS
117
34.669
2.946
52.951
1.00
45.61
B


2489
O
HIS
117
34.781
3.581
51.895
1.00
45.59
B


2490
N
LEU
118
34.094
3.435
54.044
1.00
45.87
B


2491
CA
LEU
118
33.529
4.775
54.088
1.00
46.40
B


2492
CB
LEU
118
32.919
5.026
55.469
1.00
46.21
B


2493
CG
LEU
118
31.703
4.141
55.786
1.00
46.17
B


2494
CD1
LEU
118
31.313
4.277
57.258
1.00
46.06
B


2495
CD2
LEU
118
30.539
4.535
54.877
1.00
46.03
B


2496
C
LEU
118
34.555
5.854
53.746
1.00
46.94
B


2497
O
LEU
118
34.243
6.817
53.032
1.00
46.82
B


2498
N
LYS
119
35.772
5.700
54.257
1.00
47.41
B


2499
CA
LYS
119
36.820
6.667
53.958
1.00
48.19
B


2500
CB
LYS
119
38.048
6.421
54.845
1.00
48.24
B


2501
CG
LYS
119
37.827
6.759
56.311
1.00
48.48
B


2502
CD
LYS
119
39.053
6.419
57.167
1.00
48.89
B


2503
CE
LYS
119
38.859
6.868
58.620
1.00
49.08
B


2504
NZ
LYS
119
39.997
6.460
59.507
1.00
49.31
B


2505
C
LYS
119
37.184
6.505
52.480
1.00
48.60
B


2506
O
LYS
119
37.322
7.480
51.746
1.00
48.64
B


2507
N
SER
120
37.309
5.256
52.046
1.00
49.03
B


2508
CA
SER
120
37.644
4.951
50.665
1.00
49.52
B


2509
CB
SER
120
37.553
3.445
50.432
1.00
49.70
B


2510
OG
SER
120
37.802
3.131
49.075
1.00
50.40
B


2511
C
SER
120
36.742
5.669
49.665
1.00
49.72
B


2512
O
SER
120
37.223
6.277
48.709
1.00
49.97
B


2513
N
VAL
121
35.432
5.599
49.877
1.00
49.77
B


2514
CA
VAL
121
34.492
6.234
48.965
1.00
49.62
B


2515
CB
VAL
121
33.164
5.439
48.885
1.00
49.60
B


2516
CG1
VAL
121
33.443
4.016
48.403
1.00
49.53
B


2517
CG2
VAL
121
32.477
5.422
50.241
1.00
49.55
B


2518
C
VAL
121
34.203
7.678
49.347
1.00
49.56
B


2519
O
VAL
121
33.225
8.271
48.886
1.00
49.65
B


2520
N
GLY
122
35.059
8.237
50.199
1.00
49.41
B


2521
CA
GLY
122
34.902
9.620
50.617
1.00
49.04
B


2522
C
GLY
122
33.617
9.988
51.334
1.00
48.88
B


2523
O
GLY
122
33.131
11.113
51.202
1.00
48.83
B


2524
N
THR
123
33.061
9.057
52.101
1.00
48.62
B


2525
CA
THR
123
31.839
9.339
52.836
1.00
48.45
B


2526
CB
THR
123
30.636
8.568
52.257
1.00
48.43
B


2527
OG1
THR
123
30.897
7.159
52.307
1.00
48.37
B


2528
CG2
THR
123
30.391
8.986
50.804
1.00
48.46
B


2529
C
THR
123
32.002
8.979
54.306
1.00
48.42
B


2530
O
THR
123
31.298
8.115
54.833
1.00
48.20
B


2531
N
TYR
124
32.950
9.636
54.967
1.00
48.36
B


2532
CA
TYR
124
33.165
9.382
56.380
1.00
48.37
B


2533
CB
TYR
124
34.393
8.499
56.607
1.00
48.68
B


2534
CG
TYR
124
34.443
7.935
58.013
1.00
49.05
B


2535
CD1
TYR
124
33.410
7.121
58.491
1.00
49.13
B


2536
CE1
TYR
124
33.417
6.634
59.796
1.00
49.25
B


2537
CD2
TYR
124
35.492
8.246
58.880
1.00
49.15
B


2538
CE2
TYR
124
35.507
7.762
60.193
1.00
49.40
B


2539
CZ
TYR
124
34.465
6.959
60.643
1.00
49.37
B


2540
OH
TYR
124
34.458
6.493
61.941
1.00
49.50
B


2541
C
TYR
124
33.313
10.677
57.152
1.00
48.26
B


2542
O
TYR
124
34.405
11.031
57.595
1.00
48.28
B


2543
N
GLY
125
32.197
11.383
57.302
1.00
48.00
B


2544
CA
GLY
125
32.196
12.632
58.035
1.00
47.72
B


2545
C
GLY
125
31.789
12.370
59.471
1.00
47.48
B


2546
O
GLY
125
31.771
11.218
59.910
1.00
47.44
B


2547
N
ASP
126
31.455
13.425
60.206
1.00
47.17
B


2548
CA
ASP
126
31.060
13.259
61.597
1.00
46.93
B


2549
CB
ASP
126
30.799
14.613
62.260
1.00
47.09
B


2550
CG
ASP
126
32.027
15.509
62.267
1.00
47.31
B


2551
OD1
ASP
126
33.165
14.988
62.358
1.00
47.24
B


2552
OD2
ASP
126
31.842
16.738
62.191
1.00
47.53
B


2553
C
ASP
126
29.809
12.397
61.717
1.00
46.61
B


2554
O
ASP
126
29.712
11.554
62.605
1.00
46.57
B


2555
N
ALA
127
28.852
12.617
60.821
1.00
46.15
B


2556
CA
ALA
127
27.609
11.857
60.839
1.00
45.71
B


2557
CB
ALA
127
26.721
12.273
59.675
1.00
45.75
B


2558
C
ALA
127
27.884
10.361
60.781
1.00
45.32
B


2559
O
ALA
127
27.304
9.590
61.545
1.00
45.23
B


2560
N
GLU
128
28.777
9.958
59.883
1.00
44.95
B


2561
CA
GLU
128
29.122
8.550
59.733
1.00
44.76
B


2562
CB
GLU
128
29.907
8.324
58.438
1.00
44.56
B


2563
CG
GLU
128
29.083
8.501
57.157
1.00
44.48
B


2564
CD
GLU
128
28.644
9.932
56.933
1.00
44.62
B


2565
OE1
GLU
128
29.523
10.822
56.881
1.00
44.61
B


2566
OE2
GLU
128
27.424
10.175
56.809
1.00
44.46
B


2567
C
GLU
128
29.920
8.035
60.928
1.00
44.73
B


2568
O
GLU
128
29.732
6.894
61.369
1.00
44.60
B


2569
N
ALA
129
30.816
8.869
61.446
1.00
44.62
B


2570
CA
ALA
129
31.619
8.484
62.602
1.00
44.60
B


2571
CB
ALA
129
32.627
9.587
62.938
1.00
44.61
B


2572
C
ALA
129
30.677
8.243
63.780
1.00
44.49
B


2573
O
ALA
129
30.813
7.267
64.515
1.00
44.56
B


2574
N
GLU
130
29.714
9.137
63.951
1.00
44.45
B


2575
CA
GLU
130
28.739
9.011
65.026
1.00
44.48
B


2576
CB
GLU
130
27.858
10.256
65.078
1.00
45.14
B


2577
CG
GLU
130
28.598
11.486
65.547
1.00
46.35
B


2578
CD
GLU
130
27.778
12.747
65.390
1.00
47.07
B


2579
OE1
GLU
130
26.533
12.654
65.497
1.00
47.63
B


2580
OE2
GLU
130
28.379
13.830
65.179
1.00
47.40
B


2581
C
GLU
130
27.869
7.770
64.829
1.00
44.02
B


2582
O
GLU
130
27.526
7.081
65.791
1.00
43.83
B


2583
N
ALA
131
27.503
7.500
63.580
1.00
43.48
B


2584
CA
ALA
131
26.693
6.330
63.271
1.00
43.03
B


2585
CB
ALA
131
26.338
6.303
61.778
1.00
42.94
B


2586
C
ALA
131
27.486
5.080
63.649
1.00
42.74
B


2587
O
ALA
131
26.938
4.141
64.221
1.00
42.68
B


2588
N
MET
132
28.779
5.072
63.337
1.00
42.40
B


2589
CA
MET
132
29.617
3.925
63.658
1.00
42.20
B


2590
CB
MET
132
30.981
4.051
62.983
1.00
42.10
B


2591
CG
MET
132
30.948
3.863
61.473
1.00
41.85
B


2592
SD
MET
132
30.030
2.369
60.961
1.00
41.55
B


2593
CE
MET
132
31.020
1.045
61.669
1.00
41.45
B


2594
C
MET
132
29.798
3.746
65.167
1.00
42.24
B


2595
O
MET
132
30.032
2.636
65.642
1.00
42.08
B


2596
N
GLN
133
29.687
4.838
65.914
1.00
42.30
B


2597
CA
GLN
133
29.817
4.777
67.366
1.00
42.53
B


2598
CB
GLN
133
30.058
6.169
67.950
1.00
42.87
B


2599
CG
GLN
133
30.362
6.163
69.436
1.00
43.75
B


2600
CD
GLN
133
31.485
5.202
69.789
1.00
44.32
B


2601
OE1
GLN
133
32.478
5.088
69.056
1.00
45.00
B


2602
NE2
GLN
133
31.342
4.510
70.920
1.00
44.68
B


2603
C
GLN
133
28.514
4.210
67.915
1.00
42.32
B


2604
O
GLN
133
28.507
3.468
68.898
1.00
42.35
B


2605
N
LYS
134
27.409
4.573
67.276
1.00
42.03
B


2606
CA
LYS
134
26.112
4.063
67.689
1.00
41.86
B


2607
CB
LYS
134
25.005
4.728
66.878
1.00
41.92
B


2608
CG
LYS
134
23.614
4.381
67.347
1.00
42.04
B


2609
CD
LYS
134
22.575
5.239
66.649
1.00
42.11
B


2610
CE
LYS
134
21.173
4.882
67.129
1.00
42.16
B


2611
NZ
LYS
134
20.148
5.808
66.572
1.00
42.17
B


2612
C
LYS
134
26.156
2.566
67.411
1.00
41.63
B


2613
O
LYS
134
25.701
1.756
68.215
1.00
41.61
B


2614
N
PHE
135
26.730
2.219
66.263
1.00
41.35
B


2615
CA
PHE
135
26.898
.832
65.829
1.00
41.15
B


2616
CB
PHE
135
27.687
.831
64.513
1.00
41.27
B


2617
CG
PHE
135
27.818
−.518
63.857
1.00
41.32
B


2618
CD1
PHE
135
27.058
−.826
62.731
1.00
41.46
B


2619
CD2
PHE
135
28.757
−1.441
64.305
1.00
41.29
B


2620
CE1
PHE
135
27.231
−2.032
62.051
1.00
41.47
B


2621
CE2
PHE
135
28.941
−2.650
63.637
1.00
41.40
B


2622
CZ
PHE
135
28.177
−2.947
62.504
1.00
41.48
B


2623
C
PHE
135
27.672
.068
66.917
1.00
40.97
B


2624
O
PHE
135
27.218
−.966
67.422
1.00
40.60
B


2625
N
ALA
136
28.846
.590
67.271
1.00
40.77
B


2626
CA
ALA
136
29.692
−.025
68.295
1.00
40.78
B


2627
CB
ALA
136
30.960
.810
68.493
1.00
40.74
B


2628
C
ALA
136
28.966
−.182
69.634
1.00
40.78
B


2629
O
ALA
136
29.035
−1.235
70.260
1.00
40.59
B


2630
N
GLU
137
28.281
.871
70.073
1.00
40.99
B


2631
CA
GLU
137
27.557
.836
71.343
1.00
41.43
B


2632
CB
GLU
137
26.864
2.176
71.621
1.00
42.20
B


2633
CG
GLU
137
27.792
3.326
72.023
1.00
43.67
B


2634
CD
GLU
137
28.600
3.022
73.277
1.00
44.55
B


2635
OE1
GLU
137
27.989
2.739
74.335
1.00
45.15
B


2636
OE2
GLU
137
29.854
3.062
73.204
1.00
45.31
B


2637
C
GLU
137
26.514
−.278
71.396
1.00
41.11
B


2638
O
GLU
137
26.279
−.857
72.454
1.00
41.14
B


2639
N
ALA
138
25.888
−.577
70.262
1.00
40.77
B


2640
CA
ALA
138
24.858
−1.616
70.210
1.00
40.34
B


2641
CB
ALA
138
24.142
−1.581
68.856
1.00
40.48
B


2642
C
ALA
138
25.408
−3.005
70.466
1.00
40.04
B


2643
O
ALA
138
24.699
−3.863
70.986
1.00
39.85
B


2644
N
PHE
139
26.665
−3.243
70.102
1.00
39.74
B


2645
CA
PHE
139
27.269
−4.561
70.314
1.00
39.63
B


2646
CB
PHE
139
28.345
−4.853
69.260
1.00
39.43
B


2647
CG
PHE
139
27.811
−5.179
67.890
1.00
39.21
B


2648
CD1
PHE
139
27.460
−4.167
67.005
1.00
39.17
B


2649
CD2
PHE
139
27.685
−6.506
67.484
1.00
39.03
B


2650
CE1
PHE
139
26.989
−4.469
65.723
1.00
39.05
B


2651
CE2
PHE
139
27.216
−6.822
66.211
1.00
39.04
B


2652
CZ
PHE
139
26.867
−5.804
65.326
1.00
38.97
B


2653
C
PHE
139
27.918
−4.716
71.683
1.00
39.74
B


2654
O
PHE
139
28.161
−5.836
72.125
1.00
39.68
B


2655
N
LYS
140
28.210
−3.606
72.352
1.00
39.82
B


2656
CA
LYS
140
28.884
−3.681
73.650
1.00
40.23
B


2657
CB
LYS
140
29.142
−2.274
74.193
1.00
40.29
B


2658
CG
LYS
140
30.232
−1.521
73.430
1.00
40.72
B


2659
CD
LYS
140
30.644
−.267
74.150
1.00
41.15
B


2660
CE
LYS
140
31.701
.493
73.377
1.00
41.49
B


2661
NZ
LYS
140
31.992
1.789
74.052
1.00
41.94
B


2662
C
LYS
140
28.231
−4.543
74.729
1.00
40.23
B


2663
O
LYS
140
28.914
−5.361
75.355
1.00
40.34
B


2664
N
PRO
141
26.922
−4.415
74.954
1.00
40.31
B


2665
CD
PRO
141
25.967
−3.418
74.428
1.00
40.32
B


2666
CA
PRO
141
26.297
−5.235
76.005
1.00
40.41
B


2667
CB
PRO
141
25.105
−4.395
76.414
1.00
40.39
B


2668
CG
PRO
141
24.665
−3.838
75.091
1.00
40.45
B


2669
C
PRO
141
25.877
−6.623
75.540
1.00
40.40
B


2670
O
PRO
141
25.189
−7.342
76.263
1.00
40.38
B


2671
N
VAL
142
26.320
−7.000
74.349
1.00
40.41
B


2672
CA
VAL
142
25.954
−8.285
73.778
1.00
40.49
B


2673
CB
VAL
142
25.438
−8.098
72.316
1.00
40.45
B


2674
CG1
VAL
142
25.140
−9.451
71.680
1.00
40.38
B


2675
CG2
VAL
142
24.196
−7.220
72.314
1.00
40.30
B


2676
C
VAL
142
27.108
−9.278
73.754
1.00
40.66
B


2677
O
VAL
142
28.268
−8.904
73.599
1.00
40.50
B


2678
N
ASN
143
26.771
−10.551
73.919
1.00
40.87
B


2679
CA
ASN
143
27.760
−11.614
73.836
1.00
41.19
B


2680
CB
ASN
143
28.002
−12.273
75.193
1.00
41.32
B


2681
CG
ASN
143
29.095
−13.323
75.129
1.00
41.65
B


2682
OD1
ASN
143
30.185
−13.068
74.598
1.00
41.53
B


2683
ND2
ASN
143
28.816
−14.511
75.668
1.00
41.69
B


2684
C
ASN
143
27.166
−12.629
72.871
1.00
41.36
B


2685
O
ASN
143
25.950
−12.825
72.856
1.00
41.16
B


2686
N
PHE
144
28.010
−13.271
72.069
1.00
41.61
B


2687
CA
PHE
144
27.518
−14.243
71.098
1.00
42.05
B


2688
CB
PHE
144
27.889
−13.834
69.662
1.00
41.83
B


2689
CG
PHE
144
27.275
−12.542
69.202
1.00
41.67
B


2690
CD1
PHE
144
28.017
−11.364
69.198
1.00
41.68
B


2691
CD2
PHE
144
25.964
−12.509
68.735
1.00
41.57
B


2692
CE1
PHE
144
27.461
−10.166
68.730
1.00
41.53
B


2693
CE2
PHE
144
25.401
−11.328
68.270
1.00
41.42
B


2694
CZ
PHE
144
26.153
−10.149
68.266
1.00
41.44
B


2695
C
PHE
144
28.030
−15.658
71.312
1.00
42.43
B


2696
O
PHE
144
29.108
−16.011
70.833
1.00
42.44
B


2697
N
PRO
145
27.273
−16.486
72.040
1.00
42.74
B


2698
CD
PRO
145
26.074
−16.216
72.850
1.00
42.84
B


2699
CA
PRO
145
27.741
−17.857
72.241
1.00
42.99
B


2700
CB
PRO
145
26.766
−18.408
73.281
1.00
42.99
B


2701
CG
PRO
145
26.266
−17.161
73.984
1.00
43.06
B


2702
C
PRO
145
27.564
−18.516
70.875
1.00
43.22
B


2703
O
PRO
145
26.905
−17.959
69.991
1.00
43.21
B


2704
N
PRO
146
28.154
−19.697
70.669
1.00
43.41
B


2705
CD
PRO
146
28.924
−20.582
71.555
1.00
43.53
B


2706
CA
PRO
146
27.956
−20.302
69.351
1.00
43.39
B


2707
CB
PRO
146
28.683
−21.641
69.467
1.00
43.46
B


2708
CG
PRO
146
28.663
−21.930
70.936
1.00
43.66
B


2709
C
PRO
146
26.462
−20.447
69.063
1.00
43.32
B


2710
O
PRO
146
25.681
−20.780
69.954
1.00
43.39
B


2711
N
GLY
147
26.063
−20.160
67.828
1.00
43.08
B


2712
CA
GLY
147
24.658
−20.264
67.473
1.00
42.71
B


2713
C
GLY
147
23.951
−18.930
67.579
1.00
42.36
B


2714
O
GLY
147
22.958
−18.687
66.893
1.00
42.50
B


2715
N
ALA
148
24.455
−18.058
68.440
1.00
41.98
B


2716
CA
ALA
148
23.857
−16.739
68.605
1.00
41.52
B


2717
CB
ALA
148
24.539
−15.994
69.742
1.00
41.54
B


2718
C
ALA
148
24.037
−15.998
67.281
1.00
41.26
B


2719
O
ALA
148
24.953
−16.299
66.511
1.00
41.14
B


2720
N
SER
149
23.177
−15.020
67.015
1.00
40.78
B


2721
CA
SER
149
23.253
−14.316
65.749
1.00
40.27
B


2722
CB
SER
149
22.179
−14.852
64.790
1.00
40.27
B


2723
OG
SER
149
22.238
−16.259
64.676
1.00
40.39
B


2724
C
SER
149
23.087
−12.816
65.823
1.00
39.88
B


2725
O
SER
149
22.569
−12.264
66.800
1.00
39.88
B


2726
N
VAL
150
23.544
−12.169
64.761
1.00
39.35
B


2727
CA
VAL
150
23.392
−10.743
64.608
1.00
38.82
B


2728
CB
VAL
150
24.735
−9.980
64.563
1.00
38.79
B


2729
CG1
VAL
150
25.670
−10.565
63.504
1.00
38.52
B


2730
CG2
VAL
150
24.453
−8.512
64.276
1.00
38.56
B


2731
C
VAL
150
22.687
−10.599
63.274
1.00
38.60
B


2732
O
VAL
150
22.990
−11.308
62.308
1.00
38.40
B


2733
N
PHE
151
21.717
−9.703
63.232
1.00
38.32
B


2734
CA
PHE
151
20.978
−9.478
62.009
1.00
38.25
B


2735
CB
PHE
151
19.490
−9.775
62.229
1.00
38.31
B


2736
CG
PHE
151
19.165
−11.244
62.290
1.00
38.50
B


2737
CD1
PHE
151
18.844
−11.943
61.130
1.00
38.57
B


2738
CD2
PHE
151
19.196
−11.931
63.500
1.00
38.67
B


2739
CE1
PHE
151
18.554
−13.308
61.173
1.00
38.62
B


2740
CE2
PHE
151
18.908
−13.304
63.559
1.00
38.64
B


2741
CZ
PHE
151
18.587
−13.989
62.393
1.00
38.59
B


2742
C
PHE
151
21.150
−8.034
61.587
1.00
38.12
B


2743
O
PHE
151
20.826
−7.121
62.351
1.00
37.93
B


2744
N
TYR
152
21.681
−7.837
60.383
1.00
37.98
B


2745
CA
TYR
152
21.847
−6.502
59.837
1.00
37.97
B


2746
CB
TYR
152
23.189
−6.340
59.109
1.00
37.80
B


2747
CG
TYR
152
24.421
−6.563
59.959
1.00
37.59
B


2748
CD1
TYR
152
25.033
−7.812
60.017
1.00
37.45
B


2749
CE1
TYR
152
26.202
−8.006
60.756
1.00
37.50
B


2750
CD2
TYR
152
25.002
−5.512
60.667
1.00
37.53
B


2751
CE2
TYR
152
26.170
−5.696
61.408
1.00
37.41
B


2752
CZ
TYR
152
26.760
−6.941
61.442
1.00
37.42
B


2753
OH
TYR
152
27.922
−7.121
62.145
1.00
37.70
B


2754
C
TYR
152
20.720
−6.338
58.827
1.00
38.12
B


2755
O
TYR
152
20.657
−7.048
57.822
1.00
37.99
B


2756
N
ARG
153
19.818
−5.415
59.110
1.00
38.21
B


2757
CA
ARG
153
18.709
−5.153
58.223
1.00
38.55
B


2758
CB
ARG
153
17.428
−4.982
59.041
1.00
39.13
B


2759
CG
ARG
153
16.155
−4.771
58.226
1.00
39.93
B


2760
CD
ARG
153
15.067
−4.167
59.126
1.00
40.78
B


2761
NE
ARG
153
14.543
−2.975
58.488
1.00
41.84
B


2762
CZ
ARG
153
14.398
−1.795
59.070
1.00
42.20
B


2763
NH1
ARG
153
14.730
−1.609
60.341
1.00
42.29
B


2764
NH2
ARG
153
13.926
−.784
58.352
1.00
43.17
B


2765
C
ARG
153
19.056
−3.861
57.489
1.00
38.47
B


2766
O
ARG
153
18.983
−2.774
58.062
1.00
38.45
B


2767
N
GLN
154
19.459
−3.989
56.232
1.00
38.38
B


2768
CA
GLN
154
19.817
−2.830
55.432
1.00
38.51
B


2769
CB
GLN
154
20.923
−3.170
54.444
1.00
38.40
B


2770
CG
GLN
154
21.385
−1.991
53.598
1.00
38.16
B


2771
CD
GLN
154
22.381
−2.404
52.527
1.00
38.27
B


2772
OE1
GLN
154
23.205
−3.307
52.736
1.00
37.97
B


2773
NE2
GLN
154
22.319
−1.738
51.375
1.00
37.83
B


2774
C
GLN
154
18.593
−2.356
54.663
1.00
38.75
B


2775
O
GLN
154
18.205
−2.949
53.655
1.00
38.54
B


2776
N
SER
155
17.987
−1.286
55.149
1.00
39.11
B


2777
CA
SER
155
16.814
−.708
54.519
1.00
39.50
B


2778
CB
SER
155
16.042
.135
55.532
1.00
39.57
B


2779
OG
SER
155
15.119
.981
54.876
1.00
40.10
B


2780
C
SER
155
17.237
.159
53.358
1.00
39.71
B


2781
O
SER
155
18.228
.889
53.423
1.00
39.77
B


2782
N
PRO
156
16.479
.113
52.249
1.00
39.90
B


2783
CD
PRO
156
15.293
−.728
51.977
1.00
39.90
B


2784
CA
PRO
156
16.861
.946
51.099
1.00
40.02
B


2785
CB
PRO
156
16.011
.379
49.948
1.00
39.96
B


2786
CG
PRO
156
14.811
−.238
50.609
1.00
39.92
B


2787
C
PRO
156
16.629
2.434
51.368
1.00
40.13
B


2788
O
PRO
156
17.058
3.294
50.606
1.00
40.33
B


2789
N
ASP
157
15.970
2.717
52.490
1.00
40.28
B


2790
CA
ASP
157
15.667
4.081
52.914
1.00
40.49
B


2791
CB
ASP
157
14.391
4.083
53.750
1.00
40.92
B


2792
CG
ASP
157
13.192
3.535
52.985
1.00
41.43
B


2793
OD1
ASP
157
12.330
2.899
53.628
1.00
41.85
B


2794
OD2
ASP
157
13.115
3.745
51.752
1.00
41.53
B


2795
C
ASP
157
16.816
4.723
53.729
1.00
40.33
B


2796
O
ASP
157
16.605
5.720
54.410
1.00
40.34
B


2797
N
GLY
158
18.010
4.133
53.657
1.00
40.15
B


2798
CA
GLY
158
19.140
4.691
54.365
1.00
39.99
B


2799
C
GLY
158
19.208
4.408
55.854
1.00
40.01
B


2800
O
GLY
158
19.720
5.220
56.627
1.00
39.89
B


2801
N
ILE
159
18.691
3.260
56.270
1.00
39.97
B


2802
CA
ILE
159
18.712
2.894
57.678
1.00
40.14
B


2803
CB
ILE
159
17.291
2.866
58.288
1.00
40.25
B


2804
CG2
ILE
159
17.342
2.297
59.717
1.00
40.47
B


2805
CG1
ILE
159
16.699
4.270
58.296
1.00
40.50
B


2806
CD1
ILE
159
15.253
4.319
58.750
1.00
40.85
B


2807
C
ILE
159
19.299
1.508
57.827
1.00
40.03
B


2808
O
ILE
159
19.093
.652
56.974
1.00
40.19
B


2809
N
LEU
160
20.043
1.298
58.906
1.00
39.83
B


2810
CA
LEU
160
20.617
−.002
59.190
1.00
39.58
B


2811
CB
LEU
160
22.143
.065
59.282
1.00
39.62
B


2812
CG
LEU
160
22.798
−1.270
59.656
1.00
39.78
B


2813
CD1
LEU
160
22.595
−2.268
58.535
1.00
39.75
B


2814
CD2
LEU
160
24.282
−1.076
59.920
1.00
39.82
B


2815
C
LEU
160
20.044
−.429
60.535
1.00
39.57
B


2816
O
LEU
160
20.316
.194
61.571
1.00
39.38
B


2817
N
GLY
161
19.233
−1.478
60.510
1.00
39.51
B


2818
CA
GLY
161
18.649
−1.985
61.738
1.00
39.53
B


2819
C
GLY
161
19.532
−3.094
62.270
1.00
39.62
B


2820
O
GLY
161
19.982
−3.950
61.514
1.00
39.47
B


2821
N
LEU
162
19.795
−3.075
63.570
1.00
39.77
B


2822
CA
LEU
162
20.640
−4.084
64.191
1.00
39.95
B


2823
CB
LEU
162
21.800
−3.412
64.926
1.00
39.98
B


2824
CG
LEU
162
22.800
−2.611
64.088
1.00
39.93
B


2825
CD1
LEU
162
23.870
−2.025
65.010
1.00
39.86
B


2826
CD2
LEU
162
23.434
−3.519
63.046
1.00
39.77
B


2827
C
LEU
162
19.845
−4.915
65.185
1.00
40.25
B


2828
O
LEU
162
19.181
−4.371
66.061
1.00
40.06
B


2829
N
SER
163
19.923
−6.232
65.047
1.00
40.73
B


2830
CA
SER
163
19.225
−7.140
65.948
1.00
41.50
B


2831
CB
SER
163
18.017
−7.774
65.255
1.00
41.34
B


2832
OG
SER
163
17.060
−6.787
64.940
1.00
41.93
B


2833
C
SER
163
20.177
−8.229
66.394
1.00
41.90
B


2834
O
SER
163
21.069
−8.628
65.649
1.00
41.79
B


2835
N
PHE
164
19.965
−8.708
67.613
1.00
42.62
B


2836
CA
PHE
164
20.790
−9.752
68.194
1.00
43.59
B


2837
CB
PHE
164
21.629
−9.164
69.325
1.00
43.10
B


2838
CG
PHE
164
22.354
−7.909
68.939
1.00
42.72
B


2839
CD1
PHE
164
22.033
−6.694
69.532
1.00
42.56
B


2840
CD2
PHE
164
23.350
−7.939
67.966
1.00
42.53
B


2841
CE1
PHE
164
22.695
−5.517
69.157
1.00
42.44
B


2842
CE2
PHE
164
24.015
−6.770
67.587
1.00
42.40
B


2843
CZ
PHE
164
23.684
−5.560
68.186
1.00
42.30
B


2844
C
PHE
164
19.885
−10.854
68.719
1.00
44.57
B


2845
O
PHE
164
18.927
−10.593
69.440
1.00
44.62
B


2846
N
SER
165
20.192
−12.086
68.343
1.00
45.81
B


2847
CA
SER
165
19.406
−13.237
68.756
1.00
47.07
B


2848
CB
SER
165
18.740
−13.869
67.534
1.00
47.20
B


2849
OG
SER
165
18.030
−15.044
67.884
1.00
47.51
B


2850
C
SER
165
20.276
−14.274
69.450
1.00
48.03
B


2851
O
SER
165
21.477
−14.371
69.190
1.00
47.90
B


2852
N
PRO
166
19.680
−15.053
70.367
1.00
48.99
B


2853
CD
PRO
166
18.371
−14.808
70.991
1.00
49.16
B


2854
CA
PRO
166
20.412
−16.093
71.101
1.00
49.70
B


2855
CB
PRO
166
19.505
−16.375
72.303
1.00
49.59
B


2856
CG
PRO
166
18.671
−15.115
72.421
1.00
49.47
B


2857
C
PRO
166
20.562
−17.302
70.183
1.00
50.32
B


2858
O
PRO
166
21.460
−18.124
70.361
1.00
50.60
B


2859
N
ASP
167
19.658
−17.388
69.208
1.00
51.19
B


2860
CA
ASP
167
19.676
−18.454
68.210
1.00
51.86
B


2861
CB
ASP
167
18.444
−19.374
68.339
1.00
52.48
B


2862
CG
ASP
167
17.131
−18.684
67.994
1.00
53.04
B


2863
OD1
ASP
167
17.150
−17.538
67.483
1.00
53.48
B


2864
OD2
ASP
167
16.075
−19.309
68.238
1.00
53.42
B


2865
C
ASP
167
19.739
−17.856
66.803
1.00
52.05
B


2866
O
ASP
167
20.231
−16.736
66.626
1.00
52.04
B


2867
N
THR
168
19.244
−18.605
65.822
1.00
52.24
B


2868
CA
THR
168
19.275
−18.160
64.438
1.00
52.52
B


2869
CB
THR
168
19.601
−19.338
63.460
1.00
52.70
B


2870
OG1
THR
168
18.426
−20.136
63.264
1.00
52.94
B


2871
CG2
THR
168
20.742
−20.229
63.992
1.00
52.77
B


2872
C
THR
168
17.948
−17.539
64.006
1.00
52.52
B


2873
O
THR
168
17.751
−17.201
62.838
1.00
52.57
B


2874
N
SER
169
17.029
−17.390
64.949
1.00
52.53
B


2875
CA
SER
169
15.732
−16.808
64.630
1.00
52.52
B


2876
CB
SER
169
14.657
−17.285
65.624
1.00
52.63
B


2877
OG
SER
169
14.862
−16.745
66.921
1.00
52.94
B


2878
C
SER
169
15.855
−15.301
64.685
1.00
52.30
B


2879
O
SER
169
16.604
−14.771
65.501
1.00
52.37
B


2880
N
ILE
170
15.129
−14.629
63.797
1.00
52.04
B


2881
CA
ILE
170
15.123
−13.170
63.699
1.00
51.76
B


2882
CB
ILE
170
14.443
−12.701
62.387
1.00
51.87
B


2883
CG2
ILE
170
14.507
−11.174
62.267
1.00
51.94
B


2884
CG1
ILE
170
15.132
−13.338
61.176
1.00
52.02
B


2885
CD1
ILE
170
14.499
−12.952
59.862
1.00
52.12
B


2886
C
ILE
170
14.355
−12.538
64.866
1.00
51.42
B


2887
O
ILE
170
13.164
−12.793
65.032
1.00
51.55
B


2888
N
PRO
171
15.021
−11.700
65.679
1.00
51.00
B


2889
CD
PRO
171
16.459
−11.373
65.694
1.00
50.87
B


2890
CA
PRO
171
14.330
−11.065
66.804
1.00
50.58
B


2891
CB
PRO
171
15.406
−10.155
67.391
1.00
50.62
B


2892
CG
PRO
171
16.660
−10.934
67.127
1.00
50.76
B


2893
C
PRO
171
13.126
−10.278
66.289
1.00
50.24
B


2894
O
PRO
171
13.166
−9.720
65.190
1.00
50.16
B


2895
N
GLU
172
12.054
−10.237
67.074
1.00
49.84
B


2896
CA
GLU
172
10.855
−9.514
66.666
1.00
49.51
B


2897
CB
GLU
172
9.652
−9.909
67.540
1.00
50.12
B


2898
CG
GLU
172
9.453
−9.065
68.797
1.00
51.12
B


2899
CD
GLU
172
8.170
−9.422
69.548
1.00
51.84
B


2900
OE1
GLU
172
7.546
−10.461
69.213
1.00
52.26
B


2901
OE2
GLU
172
7.791
−8.667
70.477
1.00
52.28
B


2902
C
GLU
172
11.103
−8.012
66.753
1.00
48.81
B


2903
O
GLU
172
10.439
−7.224
66.083
1.00
48.69
B


2904
N
LYS
173
12.064
−7.619
67.583
1.00
48.05
B


2905
CA
LYS
173
12.407
−6.207
67.734
1.00
47.37
B


2906
CB
LYS
173
12.023
−5.696
69.126
1.00
47.74
B


2907
CG
LYS
173
10.515
−5.552
69.338
1.00
48.42
B


2908
CD
LYS
173
10.177
−4.903
70.678
1.00
48.90
B


2909
CE
LYS
173
8.668
−4.655
70.794
1.00
49.33
B


2910
NZ
LYS
173
8.276
−3.907
72.038
1.00
49.77
B


2911
C
LYS
173
13.896
−5.987
67.503
1.00
46.60
B


2912
O
LYS
173
14.715
−6.859
67.780
1.00
46.44
B


2913
N
GLU
174
14.243
−4.820
66.986
1.00
45.80
B


2914
CA
GLU
174
15.640
−4.514
66.730
1.00
45.06
B


2915
CB
GLU
174
15.769
−3.670
65.466
1.00
44.62
B


2916
CG
GLU
174
15.225
−4.386
64.245
1.00
44.00
B


2917
CD
GLU
174
15.485
−3.643
62.962
1.00
43.55
B


2918
OE1
GLU
174
14.992
−2.505
62.820
1.00
43.16
B


2919
OE2
GLU
174
16.187
−4.202
62.096
1.00
43.38
B


2920
C
GLU
174
16.224
−3.795
67.924
1.00
44.73
B


2921
O
GLU
174
15.503
−3.129
68.659
1.00
44.71
B


2922
N
ALA
175
17.529
−3.947
68.120
1.00
44.36
B


2923
CA
ALA
175
18.220
−3.322
69.242
1.00
44.03
B


2924
CB
ALA
175
19.475
−4.120
69.588
1.00
43.98
B


2925
C
ALA
175
18.590
−1.875
68.956
1.00
43.77
B


2926
O
ALA
175
18.631
−1.046
69.863
1.00
43.58
B


2927
N
ALA
176
18.866
−1.576
67.694
1.00
43.60
B


2928
CA
ALA
176
19.241
−.225
67.318
1.00
43.48
B


2929
CB
ALA
176
20.722
.006
67.623
1.00
43.40
B


2930
C
ALA
176
18.965
.065
65.848
1.00
43.40
B


2931
O
ALA
176
18.955
−.844
65.015
1.00
43.29
B


2932
N
LEU
177
18.747
1.338
65.545
1.00
43.43
B


2933
CA
LEU
177
18.489
1.784
64.186
1.00
43.57
B


2934
CB
LEU
177
17.085
2.356
64.052
1.00
43.76
B


2935
CG
LEU
177
15.951
1.368
64.264
1.00
44.08
B


2936
CD1
LEU
177
14.635
2.092
64.008
1.00
44.44
B


2937
CD2
LEU
177
16.099
.186
63.306
1.00
44.33
B


2938
C
LEU
177
19.474
2.876
63.820
1.00
43.52
B


2939
O
LEU
177
19.379
4.001
64.317
1.00
43.68
B


2940
N
ILE
178
20.415
2.553
62.951
1.00
43.26
B


2941
CA
ILE
178
21.397
3.530
62.542
1.00
42.90
B


2942
CB
ILE
178
22.727
2.836
62.291
1.00
42.96
B


2943
CG2
ILE
178
23.773
3.844
61.862
1.00
42.91
B


2944
CG1
ILE
178
23.124
2.096
63.574
1.00
43.04
B


2945
CD1
ILE
178
24.397
1.325
63.482
1.00
43.38
B


2946
C
ILE
178
20.878
4.248
61.301
1.00
42.70
B


2947
O
ILE
178
20.779
3.662
60.226
1.00
42.77
B


2948
N
GLU
179
20.516
5.513
61.471
1.00
42.39
B


2949
CA
GLU
179
19.985
6.294
60.367
1.00
42.19
B


2950
CB
GLU
179
18.908
7.241
60.880
1.00
42.69
B


2951
CG
GLU
179
17.863
6.513
61.715
1.00
43.77
B


2952
CD
GLU
179
16.730
7.415
62.146
1.00
44.44
B


2953
OE1
GLU
179
16.872
8.654
62.041
1.00
45.03
B


2954
OE2
GLU
179
15.695
6.887
62.602
1.00
45.01
B


2955
C
GLU
179
21.088
7.062
59.667
1.00
41.66
B


2956
O
GLU
179
21.371
8.216
59.987
1.00
41.68
B


2957
N
ASN
180
21.703
6.399
58.698
1.00
40.93
B


2958
CA
ASN
180
22.783
6.986
57.925
1.00
40.29
B


2959
CB
ASN
180
24.087
6.933
58.731
1.00
40.39
B


2960
CG
ASN
180
25.235
7.594
58.011
1.00
40.49
B


2961
OD1
ASN
180
25.828
7.016
57.103
1.00
40.54
B


2962
ND2
ASN
180
25.545
8.830
58.402
1.00
40.58
B


2963
C
ASN
180
22.918
6.192
56.638
1.00
39.68
B


2964
O
ASN
180
23.260
5.008
56.653
1.00
39.49
B


2965
N
LYS
181
22.633
6.852
55.523
1.00
39.22
B


2966
CA
LYS
181
22.679
6.212
54.212
1.00
38.74
B


2967
CB
LYS
181
22.402
7.237
53.111
1.00
38.86
B


2968
CG
LYS
181
22.295
6.628
51.708
1.00
38.92
B


2969
CD
LYS
181
21.143
5.616
51.645
1.00
39.15
B


2970
CE
LYS
181
20.789
5.244
50.202
1.00
39.22
B


2971
NZ
LYS
181
19.618
4.328
50.153
1.00
39.40
B


2972
C
LYS
181
23.996
5.504
53.918
1.00
38.49
B


2973
O
LYS
181
24.010
4.323
53.582
1.00
38.22
B


2974
N
ALA
182
25.099
6.233
54.046
1.00
38.19
B


2975
CA
ALA
182
26.424
5.684
53.774
1.00
38.00
B


2976
CB
ALA
182
27.494
6.760
54.011
1.00
37.96
B


2977
C
ALA
182
26.723
4.456
54.613
1.00
37.86
B


2978
O
ALA
182
27.195
3.448
54.099
1.00
37.94
B


2979
N
VAL
183
26.455
4.541
55.910
1.00
37.80
B


2980
CA
VAL
183
26.715
3.420
56.800
1.00
37.67
B


2981
CB
VAL
183
26.640
3.864
58.282
1.00
37.56
B


2982
CG1
VAL
183
26.752
2.659
59.211
1.00
37.39
B


2983
CG2
VAL
183
27.782
4.847
58.575
1.00
37.59
B


2984
C
VAL
183
25.746
2.265
56.556
1.00
37.73
B


2985
O
VAL
183
26.120
1.094
56.687
1.00
37.63
B


2986
N
SER
184
24.513
2.587
56.176
1.00
37.66
B


2987
CA
SER
184
23.512
1.549
55.943
1.00
37.80
B


2988
CB
SER
184
22.197
2.170
55.438
1.00
37.71
B


2989
OG
SER
184
22.277
2.593
54.086
1.00
37.69
B


2990
C
SER
184
23.982
.446
54.989
1.00
37.86
B


2991
O
SER
184
23.620
−.718
55.162
1.00
38.03
B


2992
N
SER
185
24.807
.791
54.001
1.00
37.91
B


2993
CA
SER
185
25.280
−.213
53.048
1.00
38.02
B


2994
CB
SER
185
25.174
.329
51.625
1.00
38.00
B


2995
OG
SER
185
26.058
1.422
51.448
1.00
38.14
B


2996
C
SER
185
26.715
−.682
53.281
1.00
38.13
B


2997
O
SER
185
27.184
−1.611
52.618
1.00
38.15
B


2998
N
ALA
186
27.410
−.051
54.221
1.00
38.18
B


2999
CA
ALA
186
28.798
−.402
54.489
1.00
38.35
B


3000
CB
ALA
186
29.361
.471
55.626
1.00
38.24
B


3001
C
ALA
186
28.993
−1.872
54.817
1.00
38.42
B


3002
O
ALA
186
29.856
−2.525
54.240
1.00
38.53
B


3003
N
VAL
187
28.195
−2.394
55.740
1.00
38.61
B


3004
CA
VAL
187
28.328
−3.791
56.131
1.00
38.82
B


3005
CB
VAL
187
27.256
−4.194
57.167
1.00
38.75
B


3006
CG1
VAL
187
27.395
−5.670
57.513
1.00
38.72
B


3007
CG2
VAL
187
27.405
−3.348
58.420
1.00
38.82
B


3008
C
VAL
187
28.257
−4.741
54.937
1.00
38.98
B


3009
O
VAL
187
29.143
−5.570
54.750
1.00
38.83
B


3010
N
LEU
188
27.214
−4.625
54.123
1.00
39.29
B


3011
CA
LEU
188
27.088
−5.510
52.966
1.00
39.62
B


3012
CB
LEU
188
25.748
−5.291
52.264
1.00
39.52
B


3013
CG
LEU
188
25.445
−6.212
51.080
1.00
39.49
B


3014
CD1
LEU
188
25.559
−7.676
51.495
1.00
39.46
B


3015
CD2
LEU
188
24.064
−5.899
50.559
1.00
39.39
B


3016
C
LEU
188
28.230
−5.270
51.984
1.00
39.94
B


3017
O
LEU
188
28.705
−6.191
51.322
1.00
39.83
B


3018
N
GLU
189
28.664
−4.020
51.894
1.00
40.45
B


3019
CA
GLU
189
29.760
−3.663
51.005
1.00
41.13
B


3020
CB
GLU
189
30.035
−2.161
51.094
1.00
41.24
B


3021
CG
GLU
189
31.115
−1.673
50.161
1.00
41.69
B


3022
CD
GLU
189
30.654
−1.591
48.721
1.00
41.85
B


3023
OE1
GLU
189
29.616
−.949
48.461
1.00
41.80
B


3024
OE2
GLU
189
31.335
−2.163
47.847
1.00
42.32
B


3025
C
GLU
189
31.020
−4.440
51.389
1.00
41.41
B


3026
O
GLU
189
31.725
−4.961
50.523
1.00
41.54
B


3027
N
THR
190
31.303
−4.518
52.689
1.00
41.79
B


3028
CA
THR
190
32.486
−5.237
53.162
1.00
42.21
B


3029
CB
THR
190
32.727
−5.026
54.673
1.00
42.24
B


3030
OG1
THR
190
31.680
−5.667
55.414
1.00
42.19
B


3031
CG2
THR
190
32.762
−3.534
55.012
1.00
42.05
B


3032
C
THR
190
32.354
−6.736
52.934
1.00
42.52
B


3033
O
THR
190
33.318
−7.478
53.076
1.00
42.62
B


3034
N
MET
191
31.159
−7.189
52.580
1.00
42.82
B


3035
CA
MET
191
30.962
−8.612
52.369
1.00
43.21
B


3036
CB
MET
191
29.681
−9.046
53.056
1.00
43.46
B


3037
CG
MET
191
29.760
−8.921
54.551
1.00
43.97
B


3038
SD
MET
191
28.164
−9.202
55.245
1.00
44.79
B


3039
CE
MET
191
28.526
−8.892
56.998
1.00
44.57
B


3040
C
MET
191
30.936
−9.060
50.915
1.00
43.35
B


3041
O
MET
191
31.417
−10.148
50.595
1.00
43.23
B


3042
N
ILE
192
30.370
−8.237
50.039
1.00
43.58
B


3043
CA
ILE
192
30.291
−8.591
48.625
1.00
43.98
B


3044
CB
ILE
192
28.853
−9.000
48.217
1.00
43.85
B


3045
CG2
ILE
192
28.439
−10.249
48.966
1.00
43.73
B


3046
CG1
ILE
192
27.882
−7.845
48.475
1.00
43.66
B


3047
CD1
ILE
192
26.485
−8.087
47.922
1.00
43.56
B


3048
C
ILE
192
30.746
−7.470
47.701
1.00
44.41
B


3049
O
ILE
192
30.623
−7.574
46.481
1.00
44.24
B


3050
N
GLY
193
31.262
−6.397
48.289
1.00
45.00
B


3051
CA
GLY
193
31.736
−5.276
47.501
1.00
45.90
B


3052
C
GLY
193
33.059
−5.577
46.821
1.00
46.58
B


3053
O
GLY
193
33.675
−6.617
47.067
1.00
46.43
B


3054
N
GLU
194
33.502
−4.655
45.975
1.00
47.35
B


3055
CA
GLU
194
34.749
−4.833
45.247
1.00
48.29
B


3056
CB
GLU
194
35.106
−3.555
44.492
1.00
48.33
B


3057
CG
GLU
194
36.206
−3.739
43.460
1.00
48.56
B


3058
CD
GLU
194
36.792
−2.418
42.979
1.00
48.66
B


3059
OE1
GLU
194
36.092
−1.381
43.061
1.00
48.60
B


3060
OE2
GLU
194
37.952
−2.416
42.506
1.00
48.79
B


3061
C
GLU
194
35.928
−5.208
46.144
1.00
48.92
B


3062
O
GLU
194
36.695
−6.121
45.827
1.00
49.02
B


3063
N
HIS
195
36.056
−4.524
47.277
1.00
49.63
B


3064
CA
HIS
195
37.189
−4.764
48.166
1.00
50.42
B


3065
CB
HIS
195
37.550
−3.468
48.880
1.00
50.71
B


3066
CG
HIS
195
37.833
−2.346
47.937
1.00
51.19
B


3067
CD2
HIS
195
37.296
−1.103
47.823
1.00
51.39
B


3068
ND1
HIS
195
38.735
−2.459
46.894
1.00
51.39
B


3069
CE1
HIS
195
38.743
−1.339
46.193
1.00
51.51
B


3070
NE2
HIS
195
37.877
−.502
46.741
1.00
51.64
B


3071
C
HIS
195
37.065
−5.914
49.144
1.00
50.80
B


3072
O
HIS
195
37.977
−6.154
49.935
1.00
50.97
B


3073
N
ALA
196
35.959
−6.644
49.098
1.00
51.08
B


3074
CA
ALA
196
35.819
−7.795
49.978
1.00
51.47
B


3075
CB
ALA
196
34.375
−8.266
50.006
1.00
51.29
B


3076
C
ALA
196
36.711
−8.858
49.335
1.00
51.73
B


3077
O
ALA
196
36.597
−9.105
48.133
1.00
51.89
B


3078
N
VAL
197
37.599
−9.477
50.106
1.00
52.04
B


3079
CA
VAL
197
38.465
−10.489
49.519
1.00
52.26
B


3080
CB
VAL
197
39.826
−10.623
50.254
1.00
52.51
B


3081
CG1
VAL
197
40.691
−11.638
49.512
1.00
52.65
B


3082
CG2
VAL
197
40.556
−9.282
50.270
1.00
52.53
B


3083
C
VAL
197
37.798
−11.851
49.479
1.00
52.23
B


3084
O
VAL
197
37.899
−12.566
48.482
1.00
52.40
B


3085
N
SER
198
37.107
−12.220
50.548
1.00
52.13
B


3086
CA
SER
198
36.429
−13.513
50.553
1.00
51.95
B


3087
CB
SER
198
35.740
−13.764
51.886
1.00
52.14
B


3088
OG
SER
198
36.684
−13.861
52.938
1.00
52.47
B


3089
C
SER
198
35.391
−13.493
49.450
1.00
51.72
B


3090
O
SER
198
34.429
−12.733
49.514
1.00
51.76
B


3091
N
PRO
199
35.577
−14.320
48.415
1.00
51.44
B


3092
CD
PRO
199
36.784
−15.079
48.038
1.00
51.39
B


3093
CA
PRO
199
34.610
−14.333
47.318
1.00
51.14
B


3094
CB
PRO
199
35.457
−14.735
46.130
1.00
51.27
B


3095
CG
PRO
199
36.344
−15.754
46.744
1.00
51.36
B


3096
C
PRO
199
33.472
−15.299
47.473
1.00
50.85
B


3097
O
PRO
199
32.655
−15.381
46.564
1.00
50.87
B


3098
N
ASP
200
33.399
−16.015
48.594
1.00
50.39
B


3099
CA
ASP
200
32.351
−17.017
48.767
1.00
49.97
B


3100
CB
ASP
200
32.499
−17.761
50.123
1.00
50.08
B


3101
CG
ASP
200
32.358
−16.837
51.338
1.00
50.18
B


3102
OD1
ASP
200
32.336
−15.598
51.174
1.00
50.31
B


3103
OD2
ASP
200
32.279
−17.357
52.479
1.00
50.28
B


3104
C
ASP
200
30.925
−16.488
48.590
1.00
49.64
B


3105
O
ASP
200
30.109
−17.110
47.898
1.00
49.46
B


3106
N
LEU
201
30.613
−15.346
49.188
1.00
49.24
B


3107
CA
LEU
201
29.267
−14.804
49.061
1.00
49.02
B


3108
CB
LEU
201
29.046
−13.648
50.044
1.00
49.14
B


3109
CG
LEU
201
28.857
−14.062
51.513
1.00
49.23
B


3110
CD1
LEU
201
28.846
−12.833
52.390
1.00
49.11
B


3111
CD2
LEU
201
27.558
−14.833
51.680
1.00
49.22
B


3112
C
LEU
201
29.010
−14.351
47.632
1.00
48.81
B


3113
O
LEU
201
27.945
−14.612
47.086
1.00
48.56
B


3114
N
LYS
202
29.994
−13.699
47.021
1.00
48.72
B


3115
CA
LYS
202
29.847
−13.225
45.648
1.00
48.73
B


3116
CB
LYS
202
31.106
−12.493
45.195
1.00
48.87
B


3117
CG
LYS
202
31.407
−11.242
45.979
1.00
49.17
B


3118
CD
LYS
202
32.650
−10.580
45.431
1.00
49.64
B


3119
CE
LYS
202
33.209
−9.574
46.415
1.00
50.07
B


3120
NZ
LYS
202
34.703
−9.507
46.342
1.00
50.51
B


3121
C
LYS
202
29.564
−14.385
44.706
1.00
48.61
B


3122
O
LYS
202
28.737
−14.277
43.804
1.00
48.42
B


3123
N
ARG
203
30.250
−15.503
44.926
1.00
48.60
B


3124
CA
ARG
203
30.058
−16.684
44.094
1.00
48.66
B


3125
CB
ARG
203
31.087
−17.753
44.450
1.00
49.17
B


3126
CG
ARG
203
32.489
−17.349
44.052
1.00
49.89
B


3127
CD
ARG
203
33.531
−18.353
44.501
1.00
50.55
B


3128
NE
ARG
203
34.872
−17.859
44.202
1.00
51.37
B


3129
CZ
ARG
203
35.996
−18.463
44.578
1.00
51.84
B


3130
NH1
ARG
203
35.945
−19.596
45.274
1.00
52.02
B


3131
NH2
ARG
203
37.174
−17.931
44.263
1.00
52.02
B


3132
C
ARG
203
28.654
−17.242
44.229
1.00
48.40
B


3133
O
ARG
203
28.025
−17.598
43.229
1.00
48.32
B


3134
N
CYS
204
28.155
−17.316
45.461
1.00
48.04
B


3135
CA
CYS
204
26.803
−17.819
45.677
1.00
47.72
B


3136
CB
CYS
204
26.424
−17.794
47.161
1.00
47.90
B


3137
SG
CYS
204
27.254
−19.022
48.186
1.00
48.49
B


3138
C
CYS
204
25.828
−16.949
44.911
1.00
47.34
B


3139
O
CYS
204
24.968
−17.452
44.206
1.00
47.26
B


3140
N
LEU
205
25.969
−15.636
45.057
1.00
46.97
B


3141
CA
LEU
205
25.089
−14.698
44.382
1.00
46.69
B


3142
CB
LEU
205
25.493
−13.267
44.742
1.00
46.40
B


3143
CG
LEU
205
25.144
−12.876
46.184
1.00
46.24
B


3144
CD1
LEU
205
25.849
−11.584
46.574
1.00
45.98
B


3145
CD2
LEU
205
23.631
−12.735
46.309
1.00
45.95
B


3146
C
LEU
205
25.091
−14.898
42.867
1.00
46.64
B


3147
O
LEU
205
24.035
−14.908
42.236
1.00
46.55
B


3148
N
ALA
206
26.277
−15.070
42.294
1.00
46.69
B


3149
CA
ALA
206
26.413
−15.278
40.855
1.00
46.88
B


3150
CB
ALA
206
27.890
−15.249
40.453
1.00
46.76
B


3151
C
ALA
206
25.789
−16.608
40.449
1.00
47.02
B


3152
O
ALA
206
25.157
−16.712
39.401
1.00
47.02
B


3153
N
ALA
207
25.950
−17.615
41.299
1.00
47.19
B


3154
CA
ALA
207
25.420
−18.944
41.024
1.00
47.39
B


3155
CB
ALA
207
26.029
−19.947
42.001
1.00
47.39
B


3156
C
ALA
207
23.898
−19.048
41.073
1.00
47.64
B


3157
O
ALA
207
23.280
−19.643
40.189
1.00
47.61
B


3158
N
ARG
208
23.285
−18.456
42.089
1.00
47.89
B


3159
CA
ARG
208
21.840
−18.568
42.240
1.00
48.31
B


3160
CB
ARG
208
21.515
−18.804
43.723
1.00
48.48
B


3161
CG
ARG
208
22.580
−18.279
44.667
1.00
48.69
B


3162
CD
ARG
208
22.741
−19.152
45.909
1.00
48.67
B


3163
NE
ARG
208
21.526
−19.153
46.710
1.00
48.74
B


3164
CZ
ARG
208
21.111
−20.176
47.448
1.00
48.57
B


3165
NH1
ARG
208
21.812
−21.300
47.504
1.00
48.42
B


3166
NH2
ARG
208
19.973
−20.072
48.114
1.00
48.59
B


3167
C
ARG
208
20.930
−17.482
41.668
1.00
48.49
B


3168
O
ARG
208
19.725
−17.699
41.540
1.00
48.40
B


3169
N
LEU
209
21.476
−16.325
41.312
1.00
48.80
B


3170
CA
LEU
209
20.622
−15.272
40.768
1.00
49.22
B


3171
CB
LEU
209
21.320
−13.909
40.848
1.00
49.09
B


3172
CG
LEU
209
20.906
−13.011
42.022
1.00
49.04
B


3173
CD1
LEU
209
19.405
−12.750
41.957
1.00
48.97
B


3174
CD2
LEU
209
21.263
−13.669
43.337
1.00
48.92
B


3175
C
LEU
209
20.123
−15.500
39.338
1.00
49.67
B


3176
O
LEU
209
18.976
−15.190
39.025
1.00
49.50
B


3177
N
PRO
210
20.971
−16.046
38.452
1.00
50.19
B


3178
CD
PRO
210
22.389
−16.414
38.618
1.00
50.27
B


3179
CA
PRO
210
20.533
−16.273
37.070
1.00
50.82
B


3180
CB
PRO
210
21.692
−17.062
36.472
1.00
50.64
B


3181
CG
PRO
210
22.879
−16.463
37.178
1.00
50.46
B


3182
C
PRO
210
19.194
−16.996
36.952
1.00
51.51
B


3183
O
PRO
210
18.291
−16.530
36.258
1.00
51.48
B


3184
N
ALA
211
19.064
−18.122
37.645
1.00
52.30
B


3185
CA
ALA
211
17.833
−18.895
37.604
1.00
53.24
B


3186
CB
ALA
211
17.960
−20.130
38.485
1.00
53.25
B


3187
C
ALA
211
16.635
−18.062
38.037
1.00
53.94
B


3188
O
ALA
211
15.554
−18.190
37.465
1.00
53.95
B


3189
N
LEU
212
16.818
−17.211
39.045
1.00
54.79
B


3190
CA
LEU
212
15.722
−16.363
39.512
1.00
55.73
B


3191
CB
LEU
212
16.049
−15.749
40.875
1.00
55.73
B


3192
CG
LEU
212
16.141
−16.680
42.083
1.00
55.92
B


3193
CD1
LEU
212
16.463
−15.855
43.322
1.00
55.87
B


3194
CD2
LEU
212
14.822
−17.431
42.271
1.00
55.95
B


3195
C
LEU
212
15.420
−15.238
38.523
1.00
56.42
B


3196
O
LEU
212
14.264
−14.901
38.286
1.00
56.37
B


3197
N
LEU
213
16.465
−14.659
37.943
1.00
57.37
B


3198
CA
LEU
213
16.291
−13.562
37.000
1.00
58.47
B


3199
CB
LEU
213
17.636
−12.897
36.713
1.00
58.27
B


3200
CG
LEU
213
18.338
−12.286
37.926
1.00
58.28
B


3201
CD1
LEU
213
19.683
−11.738
37.498
1.00
58.29
B


3202
CD2
LEU
213
17.474
−11.191
38.534
1.00
58.19
B


3203
C
LEU
213
15.650
−13.999
35.687
1.00
59.34
B


3204
O
LEU
213
14.823
−13.276
35.124
1.00
59.41
B


3205
N
ASN
214
16.027
−15.180
35.205
1.00
60.39
B


3206
CA
ASN
214
15.492
−15.684
33.943
1.00
61.55
B


3207
CB
ASN
214
16.411
−16.765
33.357
1.00
61.91
B


3208
CG
ASN
214
16.403
−18.053
34.170
1.00
62.38
B


3209
OD1
ASN
214
15.530
−18.262
35.017
1.00
62.69
B


3210
ND2
ASN
214
17.372
−18.936
33.899
1.00
62.65
B


3211
C
ASN
214
14.072
−16.233
34.067
1.00
62.15
B


3212
O
ASN
214
13.566
−16.860
33.134
1.00
62.30
B


3213
N
GLU
215
13.438
−15.999
35.213
1.00
62.82
B


3214
CA
GLU
215
12.071
−16.465
35.443
1.00
63.51
B


3215
CB
GLU
215
11.780
−16.547
36.942
1.00
63.81
B


3216
CG
GLU
215
11.453
−15.201
37.585
1.00
64.40
B


3217
CD
GLU
215
11.472
−15.263
39.107
1.00
64.72
B


3218
OE1
GLU
215
11.573
−16.381
39.661
1.00
64.95
B


3219
OE2
GLU
215
11.383
−14.193
39.751
1.00
64.97
B


3220
C
GLU
215
11.099
−15.479
34.800
1.00
63.76
B


3221
OT1
GLU
215
11.556
−14.377
34.424
1.00
63.97
B


3222
OT2
GLU
215
9.899
−15.810
34.682
1.00
63.98
B


3223
OH2
WAT
1
31.093
−4.250
43.359
1.00
38.31
W


3224
OH2
WAT
2
22.205
−3.369
72.055
1.00
41.07
W


3225
OH2
WAT
3
20.554
−.944
41.661
1.00
39.45
W


3226
OH2
WAT
4
17.896
−6.055
62.376
1.00
38.35
W


3227
OH2
WAT
5
20.405
21.269
30.480
1.00
36.21
W


3228
OH2
WAT
6
12.876
−1.564
64.212
1.00
45.53
W


3229
OH2
WAT
7
13.804
3.554
27.585
1.00
43.05
W


3230
OH2
WAT
8
24.786
−3.380
54.771
1.00
35.11
W


3231
OH2
WAT
9
28.066
1.015
49.442
1.00
45.06
W


3232
OH2
WAT
10
28.942
.733
38.701
1.00
34.28
W


3233
OH2
WAT
11
11.034
1.040
35.900
1.00
62.48
W


3234
OH2
WAT
12
11.958
−2.453
50.194
1.00
45.87
W


3235
OH2
WAT
13
21.729
10.816
38.274
1.00
38.90
W


3236
OH2
WAT
14
14.717
1.042
31.574
1.00
54.49
W


3237
OH2
WAT
15
17.148
18.726
21.425
1.00
43.77
W


3238
OH2
WAT
16
14.258
−8.167
59.218
1.00
38.95
W


3239
OH2
WAT
17
25.220
14.836
50.255
1.00
39.68
W


3240
OH2
WAT
18
31.999
−6.768
44.244
1.00
36.85
W


3241
OH2
WAT
19
18.759
−3.066
41.285
1.00
36.91
W


3242
OH2
WAT
20
33.825
−.369
44.325
1.00
46.84
W


3243
OH2
WAT
21
21.027
6.946
63.895
1.00
39.53
W


3244
OH2
WAT
22
26.841
26.528
37.222
1.00
41.75
W


3245
OH2
WAT
23
25.367
9.122
54.856
1.00
37.37
W


3246
OH2
WAT
24
25.067
.794
29.176
1.00
43.46
W


3247
OH2
WAT
25
19.710
−3.614
38.582
1.00
41.87
W


3248
OH2
WAT
26
17.814
2.392
46.303
1.00
43.24
W


3249
OH2
WAT
27
25.147
24.525
18.999
1.00
37.60
W


3250
OH2
WAT
28
27.097
13.442
48.534
1.00
41.43
W


3251
OH2
WAT
29
27.483
37.303
38.185
1.00
49.70
W


3252
OH2
WAT
30
29.357
14.737
45.872
1.00
46.90
W


3253
OH2
WAT
31
17.807
−7.527
69.022
1.00
44.13
W


3254
OH2
WAT
32
21.345
−13.823
34.901
1.00
44.32
W


3255
OH2
WAT
33
35.351
−17.312
50.704
1.00
55.78
W


3256
OH2
WAT
34
26.136
26.351
20.833
1.00
40.13
W


3257
OH2
WAT
35
32.708
−13.771
75.135
1.00
43.56
W


3258
OH2
WAT
36
12.775
2.758
37.403
1.00
45.67
W


3259
OH2
WAT
37
25.576
21.447
40.719
1.00
44.80
W


3260
OH2
WAT
38
22.179
22.412
44.230
1.00
40.57
W


3261
OH2
WAT
39
27.629
1.078
29.037
1.00
46.71
W


3262
OH2
WAT
40
15.214
−8.497
63.801
1.00
38.34
W


3263
OH2
WAT
41
33.570
−1.687
70.758
1.00
40.60
W


3264
OH2
WAT
42
31.914
−2.489
45.115
1.00
43.00
W


3265
OH2
WAT
43
23.695
2.255
70.078
1.00
43.19
W


3266
OH2
WAT
44
29.791
12.761
49.741
1.00
48.56
W


3267
OH2
WAT
45
15.071
9.940
29.979
1.00
40.83
W


3268
OH2
WAT
46
6.237
−10.540
53.081
1.00
48.79
W


3269
OH2
WAT
47
20.476
3.837
40.323
1.00
42.14
W


3270
OH2
WAT
48
29.765
5.450
42.949
1.00
41.40
W


3271
OH2
WAT
49
23.422
10.111
60.106
1.00
41.64
W


3272
OH2
WAT
50
20.860
.481
49.642
1.00
46.53
W


3273
OH2
WAT
51
5.256
8.767
49.027
1.00
55.74
W


3274
OH2
WAT
52
24.018
33.664
24.223
1.00
52.13
W


3275
OH2
WAT
53
21.760
30.910
22.406
1.00
47.05
W


3276
OH2
WAT
54
19.585
14.021
16.441
1.00
56.45
W


3277
OH2
WAT
55
20.975
−19.597
38.847
1.00
49.49
W


3278
OH2
WAT
56
13.251
1.391
60.383
1.00
54.19
W


3279
OH2
WAT
57
23.704
7.874
64.123
1.00
44.28
W


3280
OH2
WAT
58
15.256
7.236
51.233
1.00
43.50
W


3281
OH2
WAT
59
24.722
14.018
57.391
1.00
45.21
W


3282
OH2
WAT
60
26.451
.059
45.352
1.00
44.50
W


3283
OH2
WAT
61
34.911
7.400
28.623
1.00
49.87
W


3284
OH2
WAT
62
16.024
−8.090
61.241
1.00
39.36
W


3285
OH2
WAT
63
24.783
9.796
62.406
1.00
46.71
W


3286
OH2
WAT
64
31.166
−2.930
69.992
1.00
41.41
W


3287
OH2
WAT
65
10.893
−9.636
41.973
1.00
44.07
W


3288
OH2
WAT
66
28.314
−12.912
62.930
1.00
41.86
W


3289
OH2
WAT
67
24.853
17.293
49.467
1.00
38.69
W


3290
OH2
WAT
68
36.526
.983
63.657
1.00
46.65
W


3291
OH2
WAT
69
18.777
3.199
43.897
1.00
57.05
W


3292
OH2
WAT
70
28.000
−.909
33.105
1.00
44.17
W


3293
OH2
WAT
71
24.106
9.098
47.740
1.00
55.73
W


3294
OH2
WAT
72
17.808
6.338
50.880
1.00
39.96
W


3295
OH2
WAT
73
36.807
14.820
35.205
1.00
49.63
W


3296
OH2
WAT
74
14.829
11.241
23.393
1.00
43.87
W


3297
OH2
WAT
75
36.031
33.971
37.266
1.00
56.30
W


3298
OH2
WAT
76
4.952
−6.504
59.504
1.00
57.63
W


3299
OH2
WAT
77
22.846
27.968
42.941
1.00
52.34
W


3300
OH2
WAT
78
12.146
−11.809
69.643
1.00
54.90
W


3301
OH2
WAT
79
19.700
5.591
42.910
1.00
51.71
W


3302
OH2
WAT
80
15.474
.949
68.278
1.00
66.40
W


3303
OH2
WAT
81
34.987
13.359
30.852
1.00
42.96
W


3304
OH2
WAT
82
38.932
7.300
62.259
1.00
63.71
W


3305
OH2
WAT
83
35.300
12.120
60.726
1.00
66.45
W


3306
OH2
WAT
84
8.372
12.069
21.462
1.00
64.68
W


3307
OH2
WAT
85
35.004
−3.331
69.181
1.00
47.28
W


3308
OH2
WAT
86
16.178
−17.345
60.213
1.00
62.52
W


3309
OH2
WAT
87
32.218
−1.490
37.454
1.00
45.67
W


3310
OH2
WAT
88
33.650
−5.544
36.673
1.00
48.90
W


3311
OH2
WAT
89
10.839
3.447
41.407
1.00
44.17
W


3312
OH2
WAT
90
15.338
8.207
53.876
1.00
45.01
W


3313
OH2
WAT
91
30.982
−3.939
36.099
1.00
38.73
W


3314
OH2
WAT
92
28.437
−19.091
76.433
1.00
48.06
W


3315
OH2
WAT
93
29.317
−3.476
34.057
1.00
49.49
W


3316
OH2
WAT
94
27.684
10.298
48.858
1.00
55.52
W


3317
OH2
WAT
95
14.673
−23.662
53.251
1.00
49.20
W


3318
OH2
WAT
96
17.556
7.948
19.460
1.00
63.81
W


3319
OH2
WAT
97
28.042
7.556
41.706
1.00
43.72
W


3320
OH2
WAT
98
16.210
26.298
51.998
1.00
60.02
W


3321
OH2
WAT
99
23.180
12.765
59.454
1.00
48.47
W


3322
OH2
WAT
100
24.091
26.235
51.196
1.00
66.16
W


3323
OH2
WAT
101
23.826
8.978
66.633
1.00
56.80
W


3324
OH2
WAT
102
3.301
13.799
45.653
1.00
66.31
W


3325
OH2
WAT
103
33.824
9.701
27.099
1.00
60.54
W


3326
OH2
WAT
104
29.786
12.544
55.054
1.00
41.18
W


3327
OH2
WAT
105
23.227
−12.967
71.600
1.00
51.29
W


3328
OH2
WAT
106
26.114
16.990
57.406
1.00
52.43
W


3329
OH2
WAT
107
25.488
27.629
39.671
1.00
50.05
W


3330
OH2
WAT
108
22.470
30.831
40.093
1.00
57.56
W


3331
OH2
WAT
109
37.775
5.213
63.399
1.00
69.73
W


3332
OH2
WAT
110
34.301
−.031
38.944
1.00
45.56
W


3333
OH2
WAT
111
27.597
2.887
47.690
1.00
60.49
W


3334
OH2
WAT
112
17.992
3.627
67.907
1.00
58.51
W


3335
OH2
WAT
113
34.212
−2.513
48.565
1.00
47.94
W


3336
OH2
WAT
114
9.142
3.223
51.976
1.00
57.69
W


3337
OH2
WAT
115
19.903
1.345
25.038
1.00
68.54
W


3338
OH2
WAT
116
20.188
2.209
52.296
1.00
42.73
W


3339
OH2
WAT
117
28.947
5.592
51.018
1.00
40.39
W


3340
OH2
WAT
118
16.572
−11.398
70.607
1.00
46.52
W


3341
OH2
WAT
119
22.371
28.957
16.258
1.00
55.29
W


3342
OH2
WAT
120
20.120
12.380
14.024
1.00
58.21
W


3343
OH2
WAT
121
30.963
2.318
25.429
1.00
48.38
W


3344
OH2
WAT
122
21.073
3.484
45.639
1.00
56.45
W


3345
OH2
WAT
123
34.100
.809
70.341
1.00
50.51
W


3346
OH2
WAT
124
18.016
−1.735
72.545
1.00
53.41
W


3347
OH2
WAT
125
32.760
17.849
19.769
1.00
51.60
W


3348
OH2
WAT
126
20.014
35.120
27.688
1.00
68.27
W


3349
OH2
WAT
127
39.302
.999
59.401
1.00
41.72
W


3350
OH2
WAT
128
33.457
4.342
66.754
1.00
67.39
W


3351
OH2
WAT
129
11.112
−.382
62.500
1.00
67.86
W


3352
OH2
WAT
130
29.176
−20.293
78.691
1.00
51.06
W


3353
OH2
WAT
131
18.735
27.403
17.177
1.00
47.80
W


3354
OH2
WAT
132
32.904
20.389
23.167
1.00
56.27
W


3355
OH2
WAT
133
9.835
−4.472
65.449
1.00
53.12
W


3356
OH2
WAT
134
32.597
28.207
35.870
1.00
55.11
W


3357
OH2
WAT
135
29.969
−25.969
56.239
1.00
48.12
W


3358
OH2
WAT
136
36.964
−1.362
65.077
1.00
44.08
W


3359
OH2
WAT
137
10.433
10.253
21.603
1.00
54.93
W


3360
OH2
WAT
138
38.129
13.412
41.162
1.00
48.39
W


3361
OH2
WAT
139
36.704
17.146
29.103
1.00
59.58
W


3362
OH2
WAT
140
27.851
−23.129
66.111
1.00
60.12
W


3363
OH2
WAT
141
34.360
13.181
19.409
1.00
66.62
W


3364
OH2
WAT
142
20.706
−23.727
62.602
1.00
54.01
W


3365
OH2
WAT
143
39.554
2.841
34.406
1.00
68.29
W


3366
OH2
WAT
144
25.204
−19.729
34.272
1.00
57.76
W


3367
OH2
WAT
145
41.086
−23.712
68.027
1.00
72.43
W


3368
OH2
WAT
146
16.943
6.274
41.247
1.00
41.20
W


3369
OH2
WAT
147
11.542
−4.401
42.425
1.00
47.73
W


3370
OH2
WAT
148
14.946
27.656
20.304
1.00
52.17
W


3371
OH2
WAT
149
24.832
5.955
23.125
1.00
48.56
W


3372
OH2
WAT
150
30.653
9.001
16.618
1.00
66.87
W


3373
OH2
WAT
151
28.998
14.078
23.475
1.00
43.35
W


3374
OH2
WAT
152
34.368
11.875
54.060
1.00
53.46
W


3375
OH2
WAT
153
25.939
8.771
17.733
1.00
69.63
W


3376
OH2
WAT
154
31.613
13.875
44.688
1.00
60.00
W


3377
OH2
WAT
155
29.255
.100
30.971
1.00
49.76
W


3378
OH2
WAT
156
19.059
29.293
41.016
1.00
55.35
W


3379
OH2
WAT
157
36.595
−7.763
43.708
1.00
56.53
W


3380
OH2
WAT
158
20.748
−5.549
73.212
1.00
58.80
W


3381
OH2
WAT
159
38.071
−6.743
71.037
1.00
56.27
W


3382
OH2
WAT
160
11.224
17.579
57.161
1.00
48.48
W


3383
OH2
WAT
161
31.397
14.497
22.933
1.00
49.77
W


3384
OH2
WAT
162
35.502
−7.801
38.951
1.00
50.71
W


3385
OH2
WAT
163
33.161
−21.147
71.386
1.00
51.02
W


3386
OH2
WAT
164
20.286
11.119
59.449
1.00
52.24
W


3387
OH2
WAT
165
34.529
24.258
33.196
1.00
67.98
W


3388
OH2
WAT
166
12.012
4.516
25.773
1.00
50.14
W


3389
OH2
WAT
167
31.772
11.810
47.996
1.00
67.99
W


3390
OH2
WAT
168
14.912
8.442
20.411
1.00
74.18
W


3391
OH2
WAT
169
9.368
26.392
46.096
1.00
63.81
W


3392
OH2
WAT
170
28.509
2.982
51.791
1.00
46.67
W


3393
OH2
WAT
171
4.996
6.432
40.740
1.00
63.03
W


3394
OH2
WAT
172
11.588
10.669
28.048
1.00
46.74
W


3395
OH2
WAT
173
14.227
23.192
18.948
1.00
49.28
W


3396
OH2
WAT
174
18.719
11.671
18.495
1.00
71.10
W


3397
OH2
WAT
175
13.190
−20.042
67.678
1.00
58.59
W


3398
OH2
WAT
176
25.777
−23.959
67.840
1.00
62.17
W


3399
OH2
WAT
177
16.999
27.742
45.551
1.00
58.42
W


3400
OH2
WAT
178
22.249
.128
71.871
1.00
65.44
W


3401
OH2
WAT
179
35.826
−4.408
52.580
1.00
58.54
W


3402
OH2
WAT
180
25.839
.223
74.936
1.00
55.69
W


3403
OH2
WAT
181
22.433
4.944
43.941
1.00
60.42
W


3404
OH2
WAT
182
29.223
−12.148
60.548
1.00
59.86
W


3405
OH2
WAT
183
15.667
16.612
20.418
1.00
49.49
W


3406
OH2
WAT
184
24.783
−21.882
46.145
1.00
68.71
W


3407
OH2
WAT
185
25.242
19.671
50.499
1.00
44.24
W


3408
OH2
WAT
186
2.423
26.673
41.340
1.00
67.24
W


3409
OH2
WAT
187
23.280
17.623
60.422
1.00
62.91
W


3410
OH2
WAT
188
15.145
32.923
37.151
1.00
65.04
W


3411
OH2
WAT
189
20.969
10.269
20.754
1.00
52.89
W


3412
OH2
WAT
190
18.175
−17.048
58.331
1.00
53.26
W


3413
OH2
WAT
191
24.571
−26.567
52.275
1.00
58.98
W


3414
OH2
WAT
192
38.246
−9.005
57.246
1.00
61.24
W


3415
OH2
WAT
193
36.305
−11.762
46.051
1.00
63.32
W


3416
OH2
WAT
194
11.497
20.108
16.174
1.00
63.58
W


3417
OH2
WAT
195
31.584
−21.544
74.117
1.00
68.96
W


3418
OH2
WAT
196
12.043
−.744
48.184
1.00
52.26
W


3419
OH2
WAT
197
11.989
14.143
21.333
1.00
61.32
W


3420
OH2
WAT
198
12.839
1.264
29.315
1.00
69.48
W


3421
OH2
WAT
199
11.860
−15.478
45.332
1.00
65.32
W


3422
OH2
WAT
200
20.430
13.555
61.068
1.00
59.82
W


3423
OH2
WAT
201
23.643
−4.606
32.061
1.00
64.48
W


3424
OH2
WAT
202
38.517
−25.787
70.646
1.00
76.00
W


3425
OH2
WAT
203
15.297
31.145
28.851
1.00
66.25
W


3426
OH2
WAT
204
15.159
2.605
25.487
1.00
60.04
W


3427
OH2
WAT
205
28.077
23.607
35.404
1.00
67.09
W


3428
OH2
WAT
206
32.761
39.897
39.324
1.00
57.67
W


3429
OH2
WAT
207
36.641
−2.074
53.768
1.00
57.82
W


3430
OH2
WAT
208
34.353
−1.946
73.339
1.00
50.05
W


3431
OH2
WAT
209
27.589
2.824
43.260
1.00
70.02
W


3432
OH2
WAT
210
12.853
−.417
41.865
1.00
57.86
W


3433
OH2
WAT
211
26.187
23.776
48.114
1.00
57.27
W


3434
OH2
WAT
212
11.985
−1.710
44.211
1.00
51.14
W


3435
OH2
WAT
213
38.689
7.218
30.912
1.00
68.27
W


3436
OH2
WAT
214
8.583
26.431
29.422
1.00
65.31
W


3437
OH2
WAT
215
17.594
15.705
17.887
1.00
54.42
W


3438
OH2
WAT
216
1.849
20.472
37.884
1.00
57.12
W


3439
OH2
WAT
217
8.333
−8.098
42.298
1.00
62.17
W


3440
OH2
WAT
218
8.946
−.483
52.167
1.00
57.91
W


3441
OH2
WAT
219
29.333
−23.629
74.187
1.00
70.25
W


3442
OH2
WAT
220
6.604
−3.624
52.392
1.00
62.34
W


3443
OH2
WAT
221
12.983
25.059
21.917
1.00
53.69
W


3444
OH2
WAT
222
38.735
−18.849
48.176
1.00
64.01
W


3445
OH2
WAT
223
8.728
−2.184
62.901
1.00
65.88
W


3446
OH2
WAT
224
8.411
4.504
49.068
1.00
60.02
W


3447
OH2
WAT
225
31.938
15.968
55.305
1.00
58.81
W


3448
OH2
WAT
226
32.527
−20.621
52.265
1.00
60.11
W


3449
OH2
WAT
227
9.073
30.520
40.192
1.00
59.71
W


3450
OH2
WAT
228
20.976
8.564
67.698
1.00
67.06
W


3451
OH2
WAT
229
40.435
−5.021
47.043
1.00
69.13
W


3452
OH2
WAT
230
11.670
−2.733
39.421
1.00
59.51
W


3453
OH2
WAT
231
33.789
14.958
43.047
1.00
67.65
W


3454
OH2
WAT
232
35.687
3.313
26.142
1.00
74.71
W


3455
OH2
WAT
233
38.617
1.882
62.137
1.00
66.43
W


3456
OH2
WAT
234
16.871
−20.946
65.437
1.00
60.62
W


3457
OH2
WAT
235
7.222
21.869
30.683
1.00
71.14
W


3458
OH2
WAT
236
29.336
18.254
60.245
1.00
72.61
W


3459
OH2
WAT
237
5.178
1.864
39.450
1.00
60.41
W


3460
OH2
WAT
238
12.637
1.590
57.377
1.00
69.21
W


3461
OH2
WAT
239
38.338
13.267
36.482
1.00
50.89
W


3462
OH2
WAT
240
19.406
−4.325
29.852
1.00
63.56
W


3463
OH2
WAT
241
15.305
−4.423
71.898
1.00
67.37
W


3464
OH2
WAT
242
11.386
−15.025
65.042
1.00
67.69
W


3465
OH2
WAT
243
24.769
3.511
50.789
1.00
54.60
W


3466
OH2
WAT
244
8.493
−8.294
50.615
1.00
58.19
W


3467
OH2
WAT
245
34.390
2.967
64.355
1.00
70.20
W


3468
OH2
WAT
246
36.585
8.991
63.426
1.00
64.27
W


3469
OH2
WAT
247
28.778
−21.201
45.284
1.00
66.53
W


3470
OH2
WAT
248
19.928
29.712
46.272
1.00
68.79
W


3471
OH2
WAT
249
36.168
−15.643
42.609
1.00
62.80
W


3472
OH2
WAT
250
6.443
29.483
39.025
1.00
71.20
W


3473
OH2
WAT
251
14.585
1.334
34.064
1.00
64.66
W


3474
OH2
WAT
252
37.851
−23.772
62.225
1.00
71.61
W


3475
OH2
WAT
253
13.134
10.148
21.360
1.00
70.24
W


3476
OH2
WAT
254
30.268
22.816
36.964
1.00
51.51
W


3477
OH2
WAT
255
37.927
3.391
40.343
1.00
57.64
W


3478
OH2
WAT
256
42.938
10.339
36.942
1.00
66.82
W


3479
OH2
WAT
257
28.860
7.512
45.386
1.00
63.93
W


3480
OH2
WAT
258
24.155
4.617
71.176
1.00
64.96
W


3481
OH2
WAT
259
13.549
7.501
55.948
1.00
53.90
W


3482
OH2
WAT
260
12.203
−3.194
66.196
1.00
45.03
W


3483
OH2
WAT
261
39.578
9.149
32.471
1.00
64.60
W


3484
OH2
WAT
262
25.827
6.707
70.171
1.00
63.75
W


3485
OH2
WAT
263
13.063
−9.369
70.044
1.00
61.85
W


3486
OH2
WAT
264
22.769
38.693
34.425
1.00
69.39
W


3487
OH2
WAT
265
24.833
4.824
42.490
1.00
46.49
W


3488
OH2
WAT
266
25.434
32.148
40.949
1.00
70.08
W


3489
OH2
WAT
267
17.356
3.009
24.217
1.00
66.40
W


3490
OH2
WAT
268
33.379
16.174
58.540
1.00
69.39
W


3491
OH2
WAT
269
28.851
−.908
45.949
1.00
46.04
W


3492
OH2
WAT
270
14.126
−.572
35.803
1.00
64.61
W


3493
OH2
WAT
271
18.950
27.610
56.135
1.00
66.68
W


3494
OH2
WAT
272
23.906
6.480
20.580
1.00
63.73
W


3495
OH2
WAT
273
33.719
−8.571
43.438
1.00
51.28
W


3496
OH2
WAT
274
24.311
26.011
16.450
1.00
43.15
W


3497
OH2
WAT
275
21.426
20.152
32.573
1.00
47.14
W


3498
OH2
WAT
276
26.446
8.758
44.029
1.00
51.61
W


3499
OH2
WAT
277
12.889
−21.632
53.500
1.00
52.81
W


3500
OH2
WAT
278
41.069
38.032
31.898
1.00
57.66
W


3501
OH2
WAT
279
23.117
28.163
40.325
1.00
44.59
W


3502
OH2
WAT
280
26.857
12.778
56.394
1.00
49.45
W


3503
OH2
WAT
281
30.368
17.578
41.493
1.00
55.61
W


3504
OH2
WAT
282
13.016
29.422
25.555
1.00
48.14
W


3505
OH2
WAT
283
19.874
29.181
52.014
1.00
71.41
W


3506
OH2
WAT
284
28.920
16.418
43.716
1.00
51.39
W


3507
OH2
WAT
285
18.685
1.850
48.511
1.00
51.50
W


3508
OH2
WAT
286
29.129
25.983
19.126
1.00
57.44
W


3509
OH2
WAT
287
13.978
25.840
18.608
1.00
60.66
W


3510
OH2
WAT
288
33.677
−18.113
32.020
1.00
60.74
W


3511
OH2
WAT
289
22.914
−20.293
70.652
1.00
58.51
W


3512
OH2
WAT
290
31.910
−12.638
49.166
1.00
53.02
W


3513
OH2
WAT
291
27.439
24.079
17.763
1.00
49.94
W


3514
OH2
WAT
292
32.424
16.265
40.423
1.00
49.57
W


3515
OH2
WAT
293
21.240
−8.372
30.296
1.00
63.65
W


3516
OH2
WAT
294
11.765
−10.836
37.034
1.00
71.61
W


3517
OH2
WAT
295
40.879
5.315
51.721
1.00
61.78
W


3518
OH2
WAT
296
2.507
23.113
38.112
1.00
60.11
W


3519
OH2
WAT
297
11.474
1.699
43.768
1.00
55.91
W


3520
OH2
WAT
298
13.539
.130
66.634
1.00
59.35
W


3521
OH2
WAT
299
41.707
2.695
55.851
1.00
53.29
W


3522
OH2
WAT
300
18.341
−15.413
75.839
1.00
63.84
W


3523
OH2
WAT
301
12.349
10.624
30.772
1.00
57.13
W


3524
OH2
WAT
302
10.595
2.398
47.881
1.00
62.67
W


3525
OH2
WAT
303
40.012
3.605
58.475
1.00
64.07
W


3526
OH2
WAT
304
18.383
−20.177
41.953
1.00
68.55
W


3527
OH2
WAT
305
21.609
30.526
44.140
1.00
74.34
W


3528
OH2
WAT
306
14.691
28.352
47.650
1.00
70.27
W


3529
OH2
WAT
307
33.449
2.986
71.701
1.00
69.46
W


3530
OH2
WAT
308
13.414
−6.967
32.790
1.00
65.31
W


3531
OH2
WAT
309
33.466
−3.525
30.583
1.00
62.35
W


3532
OH2
WAT
310
16.536
−26.252
50.513
1.00
59.40
W


3533
OH2
WAT
311
12.868
−17.768
46.259
1.00
64.49
W


3534
OH2
WAT
312
38.794
−23.819
72.570
1.00
80.28
W


3535
OH2
WAT
313
8.998
18.417
30.866
1.00
68.82
W


3536
OH2
WAT
314
34.911
8.822
19.832
1.00
73.20
W


3537
OH2
WAT
315
8.915
−3.801
49.634
1.00
61.40
W


3538
OH2
WAT
316
38.660
11.497
31.752
1.00
68.73
W


3539
OH2
WAT
317
16.707
−21.982
69.442
1.00
60.76
W


3540
OH2
WAT
318
12.028
−10.177
28.742
1.00
70.20
W


3541
OH2
WAT
319
5.774
−5.828
54.505
1.00
65.34
W


3542
OH2
WAT
320
39.266
1.301
38.833
1.00
58.62
W


3543
OH2
WAT
321
18.343
27.444
61.597
1.00
62.10
W


3544
OH2
WAT
322
20.903
−21.561
66.845
1.00
63.88
W


3545
OH2
WAT
323
34.493
10.466
21.876
1.00
70.53
W


3546
OH2
WAT
324
15.691
−1.283
30.803
1.00
63.76
W


3547
OH2
WAT
325
26.634
8.299
68.122
1.00
62.31
W


3548
OH2
WAT
326
25.919
−3.163
33.600
1.00
59.63
W


3549
OH2
WAT
327
38.618
−14.348
44.238
1.00
73.50
W


3550
OH2
WAT
328
11.465
2.147
50.403
1.00
60.62
W


3551
OH2
WAT
329
28.096
32.068
40.827
1.00
76.59
W


3552
OH2
WAT
330
36.452
12.479
48.769
1.00
62.77
W


3553
OH2
WAT
331
30.966
.750
44.943
1.00
51.62
W


3554
OH2
WAT
332
21.804
7.976
19.945
1.00
67.83
W


3555
OH2
WAT
333
15.722
32.585
24.096
1.00
68.69
W


3556
OH2
WAT
334
21.998
−17.667
32.451
1.00
71.85
W


3557
OH2
WAT
335
16.570
28.743
18.262
1.00
55.46
W


3558
OH2
WAT
336
15.355
29.463
26.907
1.00
56.68
W


3559
OH2
WAT
337
8.856
−4.842
42.494
1.00
57.55
W


3560
OH2
WAT
338
15.673
11.399
16.679
1.00
70.26
W


3561
OH2
WAT
339
30.499
−19.038
31.070
1.00
70.97
W


3562
OH2
WAT
340
27.359
6.348
14.986
1.00
76.17
W


3563
OH2
WAT
341
30.716
22.063
48.368
1.00
68.15
W


3564
OH2
WAT
342
17.680
32.354
47.113
1.00
74.87
W


3565
OH2
WAT
343
35.483
−23.224
64.528
1.00
67.43
W


3566
OH2
WAT
344
11.697
33.674
34.831
1.00
62.19
W


3567
OH2
WAT
345
35.198
−13.847
37.632
1.00
62.48
W


3568
OH2
WAT
346
10.971
−16.646
58.283
1.00
62.08
W


3569
OH2
WAT
347
35.865
−19.311
48.930
1.00
81.00
W


3570
OH2
WAT
348
28.051
−8.076
32.451
1.00
59.05
W


3571
OH2
WAT
349
−.526
20.824
45.987
1.00
73.19
W


3572
OH2
WAT
350
8.474
8.048
19.652
1.00
64.06
W


3573
OH2
WAT
351
39.937
7.489
45.216
1.00
63.27
W


3574
OH2
WAT
352
22.499
33.878
48.685
1.00
83.81
W


3575
OH2
WAT
353
36.190
−18.010
40.240
1.00
67.93
W


3576
OH2
WAT
354
6.583
19.327
31.179
1.00
75.92
W


3577
OH2
WAT
355
26.767
−23.288
43.403
1.00
67.04
W


3578
OH2
WAT
356
34.980
17.377
43.331
1.00
79.59
W


3579
OH2
WAT
357
29.121
3.825
23.022
1.00
64.97
W


3580
OH2
WAT
358
25.233
31.185
47.412
1.00
73.20
W


3581
OH2
WAT
359
21.888
−20.887
35.998
1.00
69.44
W


3582
OH2
WAT
360
41.720
−3.130
67.884
1.00
75.17
W


3583
OH2
WAT
361
22.416
−22.509
42.179
1.00
70.22
W


3584
OH2
WAT
362
15.091
−13.586
69.147
1.00
63.87
W


3585
OH2
WAT
363
14.937
−22.535
38.211
1.00
68.35
W


3586
OH2
WAT
364
42.980
−9.148
57.776
1.00
76.86
W


3587
OH2
WAT
365
21.401
−27.138
53.645
1.00
69.72
W


3588
OH2
WAT
366
10.544
−14.952
49.512
1.00
70.21
W


3589
OH2
WAT
367
12.408
3.801
61.011
1.00
62.31
W


3590
OH2
WAT
368
17.248
3.048
21.572
1.00
73.21
W


3591
OH2
WAT
369
20.917
10.215
62.345
1.00
65.29
W


3592
OH2
WAT
370
5.740
−1.049
37.946
1.00
76.08
W


3593
OH2
WAT
371
29.669
20.684
19.121
1.00
55.22
W


3594
OH2
WAT
372
40.853
−3.980
50.418
1.00
66.08
W


3595
OH2
WAT
373
16.357
33.776
29.111
1.00
72.95
W


3596
OH2
WAT
374
36.736
−21.310
42.144
1.00
75.04
W


3597
OH2
WAT
375
36.717
−1.394
68.084
1.00
57.39
W


3598
OH2
WAT
376
12.701
25.019
63.285
1.00
69.21
W


3599
OH2
WAT
377
9.461
.079
38.572
1.00
77.06
W


3600
OH2
WAT
378
29.550
−18.594
40.998
1.00
56.42
W


3601
OH2
WAT
379
37.002
33.477
33.637
1.00
69.48
W


3602
OH2
WAT
380
17.262
−22.386
60.944
1.00
73.34
W


3603
OH2
WAT
381
7.106
−5.199
74.381
1.00
73.24
W


3604
OH2
WAT
382
6.647
15.158
23.410
1.00
70.12
W


3605
C1
NAR
501
26.934
18.860
35.807
1.00
61.49
C


3606
C2
NAR
501
25.603
18.872
36.077
1.00
61.57
C


3607
C3
NAR
501
24.640
19.829
35.382
1.00
61.65
C


3608
C4
NAR
501
25.166
20.681
34.454
1.00
61.63
C


3609
C5
NAR
501
26.686
20.701
34.114
1.00
61.58
C


3610
C6
NAR
501
27.487
19.826
34.771
1.00
61.51
C


3611
C7
NAR
501
24.264
21.692
33.683
1.00
61.64
C


3612
C8
NAR
501
25.045
22.668
32.819
1.00
61.66
C


3613
C9
NAR
501
26.289
22.040
32.224
1.00
61.75
C


3614
O1
NAR
501
27.185
21.564
33.186
1.00
61.71
C


3615
C10
NAR
501
26.971
23.031
31.318
1.00
61.82
C


3616
C11
NAR
501
27.122
24.342
31.687
1.00
61.81
C


3617
C12
NAR
501
27.816
25.322
30.758
1.00
61.88
C


3618
C13
NAR
501
28.301
24.922
29.555
1.00
61.89
C


3619
C14
NAR
501
28.134
23.466
29.145
1.00
61.92
C


3620
C15
NAR
501
27.516
22.579
29.964
1.00
61.83
C


3621
O2
NAR
501
23.058
21.704
33.750
1.00
61.67
C


3622
O3
NAR
501
23.309
19.817
35.673
1.00
61.77
C


3623
O4
NAR
501
27.748
17.997
36.444
1.00
61.35
C


3624
O5
NAR
501
28.915
25.800
28.743
1.00
62.05
C


3625
C1
NAR
502
33.796
−8.354
57.778
1.00
89.25
D


3626
C2
NAR
502
32.541
−8.861
57.702
1.00
89.23
D


3627
C3
NAR
502
32.089
−10.035
58.559
1.00
89.22
D


3628
C4
NAR
502
33.006
−10.560
59.428
1.00
89.20
D


3629
C5
NAR
502
34.440
−10.025
59.563
1.00
89.20
D


3630
C6
NAR
502
34.797
−8.975
58.766
1.00
89.23
D


3631
C7
NAR
502
32.694
−11.745
60.364
1.00
89.18
D


3632
C8
NAR
502
33.367
−11.607
61.741
1.00
89.17
D


3633
C9
NAR
502
34.687
−10.819
61.746
1.00
89.16
D


3634
O1
NAR
502
35.279
−10.623
60.468
1.00
89.18
D


3635
C10
NAR
502
35.618
−11.507
62.726
1.00
89.13
D


3636
C11
NAR
502
36.217
−12.703
62.422
1.00
89.10
D


3637
C12
NAR
502
37.149
−13.377
63.431
1.00
89.06
D


3638
C13
NAR
502
37.396
−12.810
64.648
1.00
89.05
D


3639
C14
NAR
502
36.727
−11.482
64.979
1.00
89.06
D


3640
C15
NAR
502
35.896
−10.874
64.092
1.00
89.12
D


3641
O2
NAR
502
31.989
−12.680
60.059
1.00
89.20
D


3642
O3
NAR
502
30.824
−10.521
58.461
1.00
89.25
D


3643
O4
NAR
502
34.142
−7.311
56.993
1.00
89.32
D


3644
O5
NAR
502
38.221
−13.408
65.539
1.00
89.04
D


3645
S1
SUL
601
29.329
15.428
57.928
1.00
72.96
E


3646
O1
SUL
601
28.642
16.617
57.309
1.00
73.02
E


3647
O2
SUL
601
28.473
14.847
58.999
1.00
73.02
E


3648
O3
SUL
601
30.643
15.853
58.538
1.00
73.11
E


3649
O4
SUL
601
29.610
14.436
56.850
1.00
73.00
E


3650
S1
SUL
602
35.827
−2.534
36.136
1.00
74.42
F


3651
O1
SUL
602
35.316
−1.739
37.305
1.00
74.49
F


3652
O2
SUL
602
35.956
−3.971
36.514
1.00
74.45
F


3653
O3
SUL
602
37.188
−2.019
35.723
1.00
74.53
F


3654
O4
SUL
602
34.874
−2.354
34.996
1.00
74.40
F


3655
S1
SUL
603
23.022
36.796
31.551
1.00
84.30
G


3656
O1
SUL
603
24.269
37.382
32.155
1.00
84.26
G


3657
O2
SUL
603
22.893
35.383
31.991
1.00
84.22
G


3658
O3
SUL
603
23.099
36.815
30.053
1.00
84.26
G


3659
O4
SUL
603
21.855
37.634
31.971
1.00
84.28
G


3660
S1
SUL
604
28.867
23.364
40.821
1.00
71.85
H


3661
O1
SUL
604
28.898
22.850
39.401
1.00
71.90
H


3662
O2
SUL
604
27.477
23.295
41.360
1.00
71.86
H


3663
O3
SUL
604
29.322
24.801
40.860
1.00
71.98
H


3664
O4
SUL
604
29.814
22.540
41.641
1.00
71.87
H


3665
S1
SUL
605
37.472
−10.233
53.632
1.00
90.38
I


3666
O1
SUL
605
38.162
−10.843
54.827
1.00
90.36
I


3667
O2
SUL
605
36.461
−11.177
53.085
1.00
90.34
I


3668
O3
SUL
605
38.480
−9.940
52.560
1.00
90.32
I


3669
O4
SUL
605
36.839
−8.942
54.058
1.00
90.34
I









“Structure coordinates” refers to Cartesian coordinates (x, y, and z positions) derived from mathematical equations involving Fourier synthesis as determined from patterns obtained via diffraction of a monochromatic beam of X-rays by the atoms (scattering centers) of an isomerase polypeptide in crystal form. Diffraction data are used to calculate electron density maps of repeating protein units in the crystal (unit cell). Electron density maps are used to establish the positions of individual atoms within a crystal's unit cell. The term “crystal structure coordinates” refers to mathematical coordinates derived from mathematical equations related to the patterns obtained on diffraction of a monochromatic beam of X-rays by the atoms (scattering centers) of an isomerase polypeptide (e.g., a chalcone isomerase protein molecule) in crystal form. The diffraction data are used to calculate an electron density map of the repeating unit of the crystal. The electron density maps are used to establish the positions of the individual atoms within the unit cell of the crystal. The crystal structure coordinates of an isomerase can be obtained from a chalcone isomerase protein crystal having space group P6522 (a=90.37 Å, c=352.86 Å with two molecules per asymmetric unit and a solvent content of 72%). The coordinates of the isomerase polypeptide can also be obtained by means of computational analysis.
The term “selenomethionine substitution” refers to the method of producing a chemically modified form of the crystal of an isomerase (e.g., a chalcone isomerase). The isomerase protein is expressed by bacteria in media that is depleted in methionine and supplement with selenomethionine. Selenium is thereby incorporated into the crystal in place of methionine sulfurs. The location(s) of selenium are determined by X-ray diffraction analysis of the crystal. This information is used to generate the phase information used to construct a three-dimensional structure of the protein.
“Heavy atom derivatization” refers to a method of producing a chemically modified form of an isomerase crystal. In practice, a crystal is soaked in a solution containing heavy atom salts or organometallic compounds, e.g., lead chloride, gold thiomalate, thimerosal, uranyl acetate, and the like, which can diffuse through the crystal and bind to the protein's surface. Locations of the bound heavy atoms can be determined by X-ray diffraction analysis of the soaked crystal. This information is then used to construct phase information which can then be used to construct three-dimensional structures of the enzyme as described in Blundel, T. L., and Johnson, N. L., Protein Crystallography, Academic Press (1976), which is incorporated herein by reference.
“Unit cell” refers to a basic parallelepiped shaped block. Regular assembly of such blocks may construct the entire volume of a crystal. Each unit cell comprises a complete representation of the unit pattern, the repetition of which builds up the crystal.
“Space Group” refers to the arrangement of symmetry elements within a crystal.
“Molecular replacement” refers to generating a preliminary model of an isomerase whose structural coordinates are unknown, by orienting and positioning a molecule whose structural coordinates are known within the unit cell of the unknown crystal 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. This in turn can be subject to any of the several forms of refinement to provide a final, accurate structure of the unknown crystal (Lattman, E., 1985, in Methods in Enzymology, 11 5.55-77; Rossmann, M G., ed., “The Molecular Replacement Method” 1972, Int, Sci. Rev. Ser., No. 13, Gordon & Breach, New York). Using structure coordinates of the isomerase provided herein, molecular replacement may be used to determine the structural coordinates of a crystalline mutant, homologue, or a different crystal form of an isomerase.
“Substrate” refers to chalcone and 6′ deoxychalcone that are acted on by the chalcone isomerases and mutants thereof disclosed herein, and the like.
“Altered substrate specificity” includes a change in the ability of a mutant isomerase to produce a flavonoid product as compared to a non-mutated isomerase. Altered substrate specificity may include the ability of an isomerase to exhibit different enzymatic parameters relative to a non-mutated isomerase (Km, Vmax, etc.), use different substrates, and/or produce products that are different from those of known non-native isomerases.
A polypeptide is a chain of amino acids, regardless of length or post-translational modification (e.g., glycosylation or phosphorylation). A polypeptide or protein refers to a polymer in which the monomers are amino acid residues, which are joined together through amide bonds. When the amino acids are alpha-amino acids, either the L-optical isomer or the D-optical isomer can be used, the L-isomers being typical. An isomerase polypeptide of the invention is intended to encompass an amino acid sequence as set forth in SEQ ID NO:1 (see, Table 2) or SEQ ID NO:1 having one or more mutations, mutants, variants and conservative substitutions thereof comprising L- or D-amino acids and include modified sequences such as glycoproteins.








TABLE 2







(SEQ ID NO: 1)









maasitaitv enleypavvt spvtgksyfl ggagerglti egnfikftai gvylediava



 


slaakwkgks seelletldf yrdiisgpfe klirgskire lsgpeysrkv mencvahlks


 


vgtygdaeae amqkfaeafk pvnfppgasv fyrqspdgil glsfspdtsi pekeaalien


 


kavssavlet migehavspd lkrclaarlp allnegafki gn









Accordingly, the polypeptides of the invention are intended to cover naturally occurring proteins, as well as those which are recombinantly or synthetically synthesized. Polypeptide or protein fragments are also encompassed by the invention. Fragments can have the same or substantially the same amino acid sequence as the naturally occurring protein. A polypeptide or peptide having substantially the same sequence means that an amino add sequence is largely, but not entirely, the same, but retains a functional activity of the sequence to which it is related. In general polypeptides of the invention include peptides, or full-length protein, that contains substitutions, deletions, or insertions into the protein backbone, that would still have an approximately 70%-90% homology to the original protein over the corresponding portion. A yet greater degree of departure from homology is allowed if like-amino acids, i.e. conservative amino acid substitutions, do not count as a change in the sequence.
A polypeptide may be substantially related but for a conservative variation, such polypeptides being encompassed by the invention. A conservative variation denotes the replacement of an amino acid residue by another, biologically similar residue. Examples of conservative variations include the substitution of one hydrophobic residue such as isoleucine, valine, leucine or methionine for another, or the substitution of one polar residue for another, such as the substitution of arginine for lysine, glutamic for aspartic acids, or glutamine for asparagine, and the like. Other illustrative examples of conservative substitutions include the changes of: alanine to serine; arginine to lysine; asparagine to glutamine or histidine; aspartate to glutamate; cysteine to serine; glutamine to asparagine; glutamate to aspartate; glycine to proline; histidine to asparagine or glutamine; isoleucine to leucine or valine; leucine to valine or isoleucine; lysine to arginine, glutamine, or glutamate; methionine to leucine or isoleucine; phenylalanine to tyrosine, leucine or methionine; serine to threonine; threonine to serine; tryptophan to tyrosine; tyrosine to tryptophan or phenylalanine; valine to isoleucine or leucine, and the like. The term “conservative variation” also includes the use of a substituted amino acid in place of an unsubstituted parent amino acid provided that antibodies raised to the substituted polypeptide also immunoreact with the unsubstituted polypeptide.
Modifications and substitutions are not limited to replacement of amino acids. For a variety of purposes, such as increased stability, solubility, or configuration concerns, one skilled in the art will recognize the need to introduce other modifications, for example, deletion(s), replacement(s) or addition(s). Examples of such other modifications include incorporation of rare amino adds, dextra-amino acids, glycosylation sites, cytosine for specific disulfide bridge formation. The modified peptides can be chemically synthesized, or the isolated gene can be subjected to site-directed mutagenesis, or a synthetic gene can be synthesized and expressed in bacteria, yeast, baculovirus, tissue culture and so on.
Polypeptides of the invention include isomerase polypeptides (e.g., chalcone isomerase) from any number of plants, prokaryotes, eukaryotes, including, for example, invertebrates, mammals and humans and include sequences as set forth in SEQ ID NO:1 through SEQ ID NO:8, as well as sequences that have at least 70% homology to the sequence of SEQ ID NO:1 through SEQ ID NO:8, fragments, variants, or conservative substitutions of any of the foregoing sequences.
The term “variant” refers to polypeptides which are modified at one or more amino acid residues yet still retain the biological activity of an isomerase polypeptide. Variants can be produced by any number of means known in the art, including methods such as, for example, error-prone PCR, shuffling, oligonucleotide-directed mutagenesis, assembly PCR, sexual PCR mutagenesis, and the like, as well as any combination thereof.
By “substantially identical” is meant a polypeptide or nucleic acid exhibiting at least 50%, preferably 85%, more preferably 90%, and most preferably 95% homology to a reference amino acid or nucleic acid sequence.
Homology or identity is often measured using sequence analysis software (e.g., Sequence Analysis Software Package of the Genetics Computer Group, University of Wisconsin Biotechnology Center, 1710 University Avenue, Madison, Wis. 53705). Such software matches similar sequences by assigning degrees of homology to various deletions, substitutions and other modifications. The terms “homology” and “identity” in the context of two or more nucleic acids or polypeptide sequences, refer to two or more sequences or subsequences that are the same or have a specified percentage of amino add residues or nucleotides that are the same when compared and aligned for maximum correspondence over a comparison window or designated region as measured using any number of sequence comparison algorithms or by manual alignment and visual inspection.
For sequence comparison, typically one sequence acts as a reference sequence, to which test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percent sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
A “comparison window”, as used herein, includes reference to a segment of any one of the number of contiguous positions selected from the group consisting of from 20 to 600, usually about 50 to about 200, more usually about 100 to about 150 in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequence for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482, 1981, by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443, 1970, by the search for similarity method of Person & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444, 1988, by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection. Other algorithms for determining homology or identity include, for example, in addition to a BLAST program (Basic Local Alignment Search Tool at the National Center for Biological Information), ALIGN, AMAS (Analysis of Multiply Aligned Sequences), AMPS (Protein Multiple Sequence Alignment), ASSET (Aligned Segment Statistical Evaluation Tool), BANDS, BESTSCOR, BIOSCAN (Biological Sequence Comparative Analysis Node), BLIMPS (BLocks IMProved Searcher), FASTA, Intervals & Points, BMB, CLUSTAL V, CLUSTAL W, CONSENSUS, LCONSENSUS, WCONSENSUS, Smith-Waterman algorithm, DARWIN, Las Vegas algorithm, FNAT (Forced Nucleotide Alignment Tool), Framealign, Framesearch, DYNAMIC, FILTER, FSAP (Fristensky Sequence Analysis Package), GAP (Global Alignment Program), GENAL, GIBBS, GenQuest, ISSC (Sensitive Sequence Comparison), LALIGN (Local Sequence Alignment), LCP (Local Content Program), MACAW (Multiple Alignment Construction & Analysis Workbench), MAP (Multiple Alignment Program), MBLKP, MBLKN, PIMA (Pattern-Induced Multi-sequence Alignment), SAGA (Sequence Alignment by Genetic Algorithm) and WHAT-IF. Such alignment programs can also be used to screen genome databases to identify polynucleotide sequences having substantially identical sequences. A number of genome databases are available, for example, a substantial portion of the human genome is available as part of the Human Genome Sequencing Project (J. Roach, available on the World Wide Web at weber.u.Washington.edu/ ̃roach/human_genome_progress 2.html) (Gibbs, 1995). At least twenty-one other genomes have already been sequenced, including, for example, M. genitalium (Fraser et al., 1995), M. jannaschii (Bult et al., 1996), H. influenzae (Fleischmann et al., 1995), E. coli (Blattner et al., 1997), and yeast (S. cerevisiae) (Mewes et al., 1997), and D. melanogaster (Adams et al., 2000). Significant progress has also been made in sequencing the genomes of model organism, such as mouse, C. elegans, and Arabadopsis sp. Several databases containing genomic information annotated with some functional information are maintained by different organization, and are accessible via the internet, for example, on the World Wide Web at tigr.org/tdb, genetics.wisc.edu, standford.edu/ ̃ball, hiv-web.lanl.gov, ncbi.nlm.nih.gov, ebi.ac.uk, Pasteur.fr/other/biology, and genome.wi.mit.edu.
One example of a useful algorithm is BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402, 1977, and Altschul et al., J. Mol. Biol. 215:403-410, 1990, respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (available on the World Wide Web at ncbi.nlm.nih.gov). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighborhood word score threshold (Altschul et al., supra). These initial neighborhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) of 10, M=5, N=−4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectations (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915, 1989) alignments (B) of 50, expectation (E) of 10, M=5, N=−4, and a comparison of both strands.
The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Natl. Acad. Sci. USA 90:5873, 1993). One measure of similarity provided by BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a references sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.
In one embodiment, protein and nucleic acid sequence homologies are evaluated using the Basic Local Alignment Search Tool (“BLAST”) In particular, five specific BLAST programs are used to perform the following task:

    
    
        (1) BLASTP and BLAST3 compare an amino acid query sequence against a protein sequence database;
        (2) BLASTN compares a nucleotide query sequence against a nucleotide sequence database;
        (3) BLASTX compares the six-frame conceptual translation products of a query nucleotide sequence (both strands) against a protein sequence database;
        (4) TBLASTN compares a query protein sequence against a nucleotide sequence database translated in all six reading frames (both strands); and
        (5) TBLASTX compares the six-frame translations of a nucleotide query sequence against the six-frame translations of a nucleotide sequence database.
    
    


The BLAST programs identify homologous sequences by identifying similar segments, which are referred to herein as “high-scoring segment pairs,” between a query amino or nucleic acid sequence and a test sequence which is preferably obtained from a protein or nucleic acid sequence database. High-scoring segment pairs are preferably identified (i.e., aligned) by means of a scoring matrix, many of which are known in the art. Preferably, the scoring matrix used is the BLOSUM62 matrix (Gonnet et al., Science 256:1443-1445, 1992; Henikoff and Henikoff, Proteins 17:49-61, 1993). Less preferably, the PAM or PAM250 matrices may also be used (see, e.g., Schwartz and Dayhoff, eds., 1978, Matrices for Detecting Distance Relationships: Atlas of Protein Sequence and Structure, Washington: National Biomedical Research Foundation). BLAST programs are accessible through the U.S. National Library of Medicine, e.g., on the World Wide Web at ncbi.nlm.nih.gov.
The parameters used with the above algorithms may be adapted depending on the sequence length and degree of homology studied. In some embodiments, the parameters may be the default parameters used by the algorithms in the absence of instructions from the user.
One aspect of the invention resides in obtaining crystals of the isomerase polypeptide (e.g., chalcone isomerase) of sufficient quality to determine the three dimensional (tertiary) structure of the protein by X-ray diffraction methods. The knowledge obtained concerning the three-dimensional structure of chalcone isomerase can be used in the determination of the three dimensional structure of other isomerase polypeptides in the polyketide synthesis or flavonoid pathway. The structural coordinates of chalcone isomerase can be used to develop new isomerase enzymes or isomerase binding agents (e.g., inhibitors or substrates) using various computer models. Based on the structural coordinates of the chalcone isomerase polypeptide (e.g., the three dimensional protein structure), as described herein, novel isomerases can be engineered. In addition, small molecules which mimic or are capable of interacting with a functional domain of an isomerase polypeptide, can be designed and synthesized to modulate chalcone isomerase and other isomerase biological functions as well as the biological functions of other flavanone-related isomerases. Accordingly, in one embodiment, the invention provides a method of “rational” enzyme or drug design.
Another approach to “rational” enzyme or drug design is based on a lead compound that is discovered using high throughput screens; the lead compound is further modified based on a crystal structure of the binding regions of the molecule in question. Accordingly, another aspect of the invention is to provide related protein sequences or material which is a starting material in the rational design of new isomerases or drugs which lead to the synthesis of new flavonoids or modify the flavonoid pathway.
The present invention relates to crystallized isomerases and mutants thereof from which the position of specific α-carbon atoms and R-groups associated therewith comprising the active site can be determined in three-dimensional space. The invention also relates to structural coordinates of said chalcone isomerases, use of said structural coordinates to develop structural information related to isomerase homologues, mutants, and the like, and to crystal forms of such isomerases. Furthermore, the invention, as disclosed herein, provides a method whereby the α-carbon structural coordinates specifically determined for atoms comprising the active site of the isomerase can be used to develop isomerases wherein R-groups associated with active site α-carbon atoms are different from the R-groups found in native CHI, e.g., are mutant isomerases. In addition, the present invention provides for production of mutant chalcone isomerases based on the structural information provided herein and for use of the mutant isomerases to make a variety of flavonoid or polyketide compounds using a variety of substrates.
The present invention further provides, for the first time, crystal isomerases, as exemplified by chalcone isomerase (CHI; PDB Accession No. 1EYP, SEQ ID NOs:9-11) see Table 1 for coordinates of native CHI (SEQ ID NO:1).
Also provided are coordinates for crystals which are grown in the presence and absence of product and product analogues, thus allowing definition of the structural or atomic coordinates associated therewith. The structural coordinates allow determination of the α-carbon atoms comprising the active site, R-groups associated therewith, and the interaction of said α-carbons and said R-groups with each other. For example, CHI (SEQ ID NO:9) was co-crystallized with naringenin as a complex. Other crystallized complexes include CHI (SEQ ID NO:9) complexed with 5-deoxyflavanone and CHI (SEQ ID NO:9) complexed with 5,4′-dideoxyflavanone (PDB accession numbers 1FM7 and 1FM8, respectively, which were deposited on Aug. 16, 2000) and CHI (SEQ ID NO:9) complexed with 4′-dihydroxyflavanone (Accession No. 1JEP, Protein Data Bank). The crystals of CHI•naringenin belong to space group P6522 having unit cell dimensions of a=89.47 Å; c=351.19 Å, α=β=90°, γ=120° with a single monomer per asymmetric unit.
Crystal structures are preferably obtained at a resolution of about 1.56 angstroms to about 3 angstroms for an isomerase in the presence and in the absence of bound product or product analog. Coordinates for an isomerase in the absence of a substrate bound in the active site have been deposited at the Brookhaven National Laboratory Protein Data Bank, accession number 1EYP (SEQ ID NOs:9-11). Those skilled in the art understand that a set of structure coordinates determined by X-ray crystallography is not without standard error. Therefore, for the purpose of this invention, any set of structure coordinates wherein the active site α-carbons of an isomerase, isomerase homologue, or mutants thereof, have a root mean square deviation less than ±2.3 angstroms when superimposed using the structural coordinates listed in Table 1 and PDB Accession No. 1EYP, shall be considered identical.
CHI is a functional monomer of approximately 220 residues and has been isolated from a variety of higher plants. The overall structure of CHI resembles an upside-down bouquet that adopts an open-faced β-sandwich fold (FIGS. 1B and 1C). A large β-sheet β3a-β3f) and a layer of α-helices (α1-α7) comprise the core structure with three short β-strands (β1a, β1b, β2) on the opposite side of the large β-sheet. A search of the Protein Data Bank using DALI revealed no other structurally homologous folds. In addition, a PSI-BLAST search of sequence databases showed that CHI-like sequences are typically found in plants and these sequences display no detectable homology with other proteins. Amino acid sequence comparison of CHIs (SEQ ID NOs:1-8) from a variety of advanced land plants reveals high homology (49% to 82% amino acid sequence identity) with regions of conservation spread uniformly throughout the primary structure (FIG. 1D). Interestingly, there is conservation of residues spanning β3a, β3b, α4, and α6 in the three-dimensional structure among CHIs from different species. Notably, these structural elements form the active site on the protein surface.
The active site α-carbons of chalcone isomerase generally are not all contiguous, i.e., are not adjacent to one another in the primary amino acid sequence of the isomerase due to intervening amino acid residues between various active site α-carbons. Nevertheless, it should be appreciated that certain active site α-carbons can be adjacent to one another in some instances.
An appropriate combination of R-groups, linked to active site α-carbons, can facilitate the formation of one or more desired reaction products. The combination of R-groups selected for use in an isomerase can be any combination other than the ordered arrangements of R-groups found in known native isomerases. Typically, R-groups found on active site α-carbons are those found in naturally occurring amino acids. In some embodiments, however, R-groups other than those found in naturally occurring amino acids can be used.
The present invention permits the use of molecular design techniques to design, select, and synthesize genes encoding mutant isomerases and chalcone isomerases that produce different and/or novel flavonoid compounds using various substrates. Mutant proteins of the present invention and nucleic acids encoding the same can be designed by genetic manipulation based on structural information provided herein for the first time regarding isomerases. For example, one or more R-groups associated with the active site α-carbon atoms of CHI can be changed by altering the nucleotide sequence of the corresponding CHI gene, thus making one or more mutant chalcone isomerases. Such genetic manipulations can be guided by structural information concerning the R-groups found in the active site α-carbons when substrate is bound to the protein upon crystallization.
Mutant proteins of the present invention may be prepared in a number of ways available to the skilled artisan. For example, the gene encoding wild-type CHI may be mutated at those sites identified herein as corresponding to amino acid residues identified in the active site by means currently available to the artisan skilled in molecular biology techniques. Such techniques include oligonucleotide-directed mutagenesis, deletion, chemical mutagenesis, and the like. The protein encoded by the mutant gene is then produced by expressing the gene in, for example, a bacterial or plant expression system.
Alternatively, isomerase mutants may be generated by site specific-replacement of a particular amino acid with an unnaturally occurring amino acid or mimetic. As such, isomerase mutants may be generated through replacement of an amino acid residue or a particular cysteine or methionine residue with selenocysteine or selenomethionine. This may be achieved by growing a host organism capable of expressing either the wild-type or mutant polypeptide on a growth medium depleted of natural cysteine or methionine or both and growing on medium enriched with either selenocysteine, selenomethionine, or both. These and similar techniques are described in Sambrook et al., (Molecular Cloning, A Laboratory Manual, 2nd Ed. (1989) Cold Spring Harbor Laboratory Press).
Another suitable method of creating mutant isomerases of the present invention is based on a procedure described in Noel and Tsal, J. Cell. Biochem., 40:309-320, 1989. In so doing, the nucleic acids encoding the isomerase can be synthetically produced using oligonucleotides having overlapping regions, said oligonucleotides being degenerate at specific bases so that mutations are induced.
According to the present invention, nucleic acid sequences encoding a mutated polyketide isomerase can be produced by the methods described herein, or any alternative methods available to the skilled artisan. In designing the nucleic acid sequence of interest, it may be desirable to reengineer the gene for improved expression in a particular expression system. For example, it has been shown that many bacterially derived genes do not express well in plant systems. In some cases, plant-derived genes do not express well in bacteria. This phenomenon may be due to the non-optimal G+C content and/or A+T content of said gene relative to the expression system being used. For example, the very low G+C content of many bacterial genes results in the generation of sequences mimicking or duplicating plant gene control sequences that are highly A+T rich. The presence of A+T rich sequences within the genes introduced into plants (e.g., TATA box regions normally found in promoters) may result in aberrant transcription of the gene(s). In addition, the presence of other regulatory sequences residing in the transcribed mRNA (e.g. polyadenylation signal sequences (AAUAAA) or sequences complementary to small nuclear RNAs involved in pre-mRNA splicing) may lead to RNA instability. Therefore, one goal in the design of genes is to generate nucleic acid sequences that have a G+C content that affords mRNA stability and translation accuracy for a particular expression system.
Due to the plasticity afforded by the redundancy of the genetic code (i.e., some amino acids are specified by more than one codon), evolution of the genomes of different organisms or classes of organisms has resulted in differential usage of redundant codons. This “codon bias” is reflected in the mean base composition of protein coding regions. For example, organisms with relatively low G+C contents utilize codons having A or T in the third position of redundant codons, whereas those having higher G+C contents utilize codons having G or C in the third position. Therefore, in reengineering genes for expression, one may wish to determine the codon bias of the organism in which the gene is to be expressed. Looking at the usage of the codons as determined for genes of a particular organism deposited in GenBank can provide this information. After determining the bias thereof, the new gene sequence can be analyzed for restriction enzyme sites as well as other sites that could affect transcription such as exon:intron junctions, polyA addition signals, or RNA polymerase termination signals.
Genes encoding isomerases, such as chalcone isomerase, can be placed in an appropriate vector, depending on the artisan's interest, and can be expressed using a suitable expression system. An expression vector, as is well known in the art, typically includes elements that permit replication of said vector within the host cell and may contain one or more phenotypic markers for selection of cells containing the gene. The expression vector will typically contain sequences that control expression such as promoter sequences, ribosome binding sites, and translational initiation and termination sequences. Expression vectors may also contain elements such as subgenomic promoters, a repressor gene or various activator genes. The artisan may also choose to include nucleic acid sequences that result in secretion of the gene product, movement of said product to a particular organelle such as a plant plastid (see U.S. Pat. Nos. 4,762,785; 5,451,513 and 5,545,817, which are incorporated herein by reference) or other sequences that increase the ease of peptide purification, such as an affinity tag.
A wide variety of expression control sequences are useful in expressing native or mutated isomerases when operably linked thereto. Such expression control sequences include, for example, the early and late promoters of SV40 for animal cells, the lac system, the trp system, major operator and promoter systems of phage S, and the control regions of coat proteins, particularly those from RNA viruses in plants. In E. coli, a useful transcriptional control sequence is the T7 RNA polymerase binding promoter, which can be incorporated into a pET vector as described by Studier et al., Methods Enzymology, 185:60-89, 1990, which is incorporated herein by reference.
For expression, a desired gene should be operably linked to the expression control sequence and maintain the appropriate reading frame to permit production of the desired isomerase. Any of a wide variety of well-known expression vectors are of use to the present invention. These include, for example, vectors comprising segments of chromosomal, non-chromosomal and synthetic DNA sequences such as those derived from SV40, bacterial plasmids including those from E. coli such as col E1, pCR1, pBR322 and derivatives thereof, pMB9, wider host range plasmids such as RP4, phage DNA such as phage S, NM989, M13, and other such systems as described by Sambrook et al., (Molecular Cloning, A Laboratory Manual, 2nd Ed. (1989) Cold Spring Harbor Laboratory Press), which is incorporated herein by reference.
A wide variety of host cells are available for expressing isomerase mutants of the present invention. Such host cells include, for example, bacteria such as E. coli, Bacillus and Streptomyces, fungi, yeast, animal cells, plant cells, insect cells, and the like. Preferred embodiments of the present invention include chalcone isomerase mutants that are expressed in E. coli or in plant cells. Said plant cells can either be in suspension culture or a transgenic plant.
In order to produce transgenic plants, vectors containing the nucleic acid construct encoding isomerases and mutants thereof are inserted into the plant genome. Preferably, these recombinant vectors are capable of stable integration into the plant genome. One variable in making a transgenic plant is the choice of a selectable marker. A selectable marker is used to identify transformed cells against a high background of untransformed cells. The preference for a particular marker is at the discretion of the artisan, but any of the selectable markers may be used along with any other gene not listed herein that could function as a selectable marker. Such selectable markers include aminoglycoside phosphotransferase gene of transposon Tn5 (Aph 11) (which encodes resistance to the antibiotics kanamycin), neomycin, G418, as well as those genes which code for resistance or tolerance to glyphosate, hygromycin, methotrexate, phosphinothricin, imidazolinones, sulfonylureas, triazolopyrimidine herbicides, such as chlorosulfuron, bromoxynil, dalapon, and the like. In addition to a selectable marker, it may be desirable to use a reporter gene. In some instances a reporter gene may be used with a selectable marker. Reporter genes allow the detection of transformed cells and may be used at the discretion of the artisan. A list of these reporter genes is provided in K. Wolsing et al., Ann. Rev. Genetics, 22:421, 1988.
The genes are expressed either by promoters expressing in all tissues at all times (constitutive promoters), by promoters expressing in specific tissues (tissue-specific promoters), promoters expressing at specific stages of development (developmental promoters), and/or promoters expressing in response to a stimulus or stimuli (inducible promoters). The choice of these is at the discretion of the artisan.
Several techniques exist for introducing foreign genes into plant cells, and for obtaining plants that stably maintain and express the introduced gene. Such techniques include acceleration of genetic material coated on a substrate directly into cells (U.S. Pat. No. 4,945,050 to Cornell): Plant cells may also be transformed using Agrobacterium technology (see, for example, U.S. Pat. No. 5,177,010 to University of Toledo, U.S. Pat. No. 5,104,310 to Texas A&M, U.S. Pat. Nos. 5,149,645, 5,469,976, 5,464,763, 4,940,838, and 4,693,976 to Schilperoot, European Patent Applications 116718, 290799, 320500 to Max Planck, European Patent Applications 604662, 627752 and U.S. Pat. No. 5,591,616 to Japan Tobacco, European Patent Applications 0267159, 0292435 and U.S. Pat. No. 5,231,011 to Ciba-Geigy, U.S. Pat. Nos. 5,463,174 and 4,762,785 to Calgene, and U.S. Pat. Nos. 5,004,863 and 5,159,135 to Agracetus). Other transformation technologies include whiskers technology (see U.S. Pat. Nos. 5,302,523 and 5,464,765 to Zeneca). Electroporation technology has also been used to transform plants (see WO 87106614 to Boyce Thompson Institute, U.S. Pat. Nos. 5,472,869 and 5,384,253 to Dakalb, and WO 92/09696 and WO 93/21335 to Plant Genetic Systems, all which are incorporated by reference). Viral vector expression systems can also be used such as those described in U.S. Pat. Nos. 5,316,931, 5,589,367, 5,811,653, and 5,866,785 to BioSource, which are incorporated herein by reference.
In addition to numerous technologies for transforming plants, the type of tissue that is contacted with the genes of interest may vary as well. Suitable tissue includes, for example, embryonic tissue, callus tissue, hypocotyl, meristem, and the like. Almost all plant tissues may be transformed during de-differentiation using the appropriate techniques described herein.
Regardless of the transformation system used, a gene encoding a mutant isomerase is preferably incorporated into a gene transfer vector adapted to express said gene in a plant cell by including in the vector an expression control sequence (e.g., a plant promoter regulatory element). In addition to plant promoter regulatory elements, promoter regulatory elements from a variety of sources can be used efficiently in plant cells to express foreign genes. For example, promoter regulatory elements of bacterial origin, such as the octopine synthase promoter, the nopaline synthase promoter, the mannopine synthase promoter, and the like, may be used. Promoters of viral origin, such as the cauliflower mosaic virus (35S and 198) are also desirable. Plant promoter regulatory elements also include ribulose-1,6-bisphosphate carboxylase small subunit promoter, beta-conglycinin promoter, phaseolin promoter, ADH promoter, heat-shock promoters, tissue specific promoters, and the like. Numerous promoters are available to skilled artisans for use at their discretion.
It should be understood that not all expression vectors and expression systems function in the same way to express the mutated gene sequences of the present invention. Neither do all host cells function equally well with the same expression system. However, one skilled in the art may make a selection among these vectors, expression control sequences, and host without undue experimentation and without departing from the scope of this invention.
Once an isomerase of the present invention is expressed, the protein obtained therefrom can be purified so that structural analysis, modeling, and/or biochemical analysis can be performed, as exemplified herein. The nature of the protein obtained can be dependent on the expression system used. For example, genes, when expressed in mammalian or other eukaryotic cells, may contain latent signal sequences that may result in glycosylation, phosphorylation, or other post-translational modifications, which may or may not alter function. Therefore, a preferred embodiment of the present invention is the expression of mutant isomerase genes in E. coli cells. Once the proteins are expressed, they can be easily purified using techniques common to the person having ordinary skill in the art of protein biochemistry, such as, for example, techniques described in Colligan at al., (1997) Current Protocols in Protein Science, Chanda, V. B., Ed., John Wiley & Sons, Inc., which is incorporated herein by reference. Such techniques often include the use of cation-exchange or anion-exchange chromatography, gel filtration-size exclusion chromatography, and the like. Another technique that may be commonly used is affinity chromatography. Affinity chromatography can include the use of antibodies, substrate analogs, or histidine residues (His-tag technology).
Once purified, mutants of the present invention may be characterized by any of several different properties. For example, such mutants may have altered active site surface charges of one or more charge units. In addition, the mutants may have altered substrate specificity or product capability relative to a non-mutated isomerase (e.g., a chalcone isomerase).
The present invention allows for the characterization of isomerase mutants by crystallization followed by X-ray diffraction. Polypeptide crystallization occurs in solutions where the polypeptide concentration exceeds it solubility maximum (i.e., the polypeptide solution is supersaturated). Such solutions may be restored to equilibrium by reducing the polypeptide concentration, preferably through precipitation of the polypeptide crystals. Often polypeptides may be induced to crystallize from supersaturated solutions by adding agents that alter the polypeptide surface charges or perturb the interaction between the polypeptide and bulk water to promote associations that lead to crystallization.
Compounds known as “precipitants” are often used to decrease the solubility of the polypeptide in a concentrated solution by forming an energetically unfavorable precipitating layer around the polypeptide molecules (Weber, Advances in Protein Chemistry, 41:1-36, 1991). In addition to precipitants, other materials are sometimes added to the polypeptide crystallization solution. These include buffers to adjust the pH of the solution and salts to reduce the solubility of the polypeptide. Various precipitants are known in the art and include the following: ethanol, 3-ethyl-2,4-pentanediol, many of the polyglycols (such as polyethylene glycol), and the like.
Commonly used polypeptide crystallization methods include the following techniques: batch, hanging drop, seed initiation, dialysis, and the like. In each of these methods, it is important to promote continued crystallization after nucleation by maintaining a supersaturated solution. In the batch method, polypeptide is mixed with precipitants to achieve supersaturation, the vessel is sealed, and set aside until crystals appear. In the dialysis method, polypeptide is retained in a sealed dialysis membrane that is placed into a solution containing precipitant Equilibration across the membrane increases the polypeptide and precipitant concentrations thereby causing the polypeptide to reach supersaturation levels.
In the preferred hanging drop technique (McPherson, J. Biot Chem, 6300-6306, 1976), an initial polypeptide mixture is created by adding a precipitant to a concentrated polypeptide solution. The concentrations of the polypeptide and precipitants are such that in this initial form, the polypeptide does not crystallize. A small drop of this mixture is placed on a glass slide that is inverted and suspended over a reservoir of a second solution. The system is then sealed. Typically, the second solution contains a higher concentration of precipitant or other dehydrating agent. The difference in the precipitant concentrations causes the protein solution to have a higher vapor pressure than the solution. Since the system containing the two solutions is sealed, an equilibrium is established, and water from the polypeptide mixture transfers to the second solution. This equilibrium increases the polypeptide and precipitant concentration in the polypeptide solution. At the critical concentration of polypeptide and precipitant, a crystal of the polypeptide will form.
Another method of crystallization involves introducing a nucleation site into a concentrated polypeptide solution. Generally, a concentrated polypeptide solution is prepared and a seed crystal of the polypeptide is introduced into this solution. If the concentration of the polypeptide and any precipitants are correct, the seed crystal will provide a nucleation site around which a larger crystal forms. In typical embodiments, the crystals of the present invention are formed in hanging drops with 15% PEG 8000; 200 mM magnesium acetate or magnesium chloride, 100 mM 3-(N-morpholino)-2-hydroxypropanesulfonic acid (pH 7.0), and 1 mM dithiothreitol as precipitant.
Some proteins may be recalcitrant to crystallization. However, several techniques are available to the skilled artisan. Quite often the removal of polypeptide segments at the amino or carboxy terminal end of the protein is necessary to produce crystalline protein samples. Said procedures involve either treatment of the protein with one of several proteases including trypsin, chymotrypsin, substilisin, and the like. This treatment often results in the removal of flexible polypeptide segments that are likely to negatively affect crystallization. Alternatively, the removal of coding sequences from the protein's gene facilitates the recombinant expression of shortened proteins that can be screened for crystallization.
The crystals so produced have a wide range of uses. For example, high quality crystals are suitable for X-ray or neutron diffraction analysis to determine the three-dimensional structure of mutant and native isomerases and to design additional mutants thereof. In addition, crystallization can serve as a further purification method. In some instances, a polypeptide or protein will crystallize from a heterogeneous mixture into crystals. Isolation of such crystals by filtration, centrifugation, etc., followed by redissolving the polypeptide affords a purified solution suitable for use in growing the high-quality crystals needed for diffraction studies. The high-quality crystals may also be dissolved in water and then formulated to provide an aqueous solution having other uses as desired.
Because isomerases may crystallize in more than one crystal form, the structural coordinates of α-carbons of an active site determined from an isomerase or portions thereof, as provided by this invention, are particularly useful to solve the structure of other crystal forms of isomerases. The structural coordinates, as provided herein, may also be used to solve the structure of isomerases having α-carbons positioned within the active sites in a manner similar to the wild-type isomerase, yet having R-groups that may or may not be identical to the wild-type isomerase.
Furthermore, the structural coordinates disclosed herein may be used to determine the structure of the crystalline form of other proteins with significant amino acid or structural homology to any functional domain of an isomerase. One method that may be employed for such purpose is molecular replacement. In this method, the unknown crystal structure, whether it is another crystal form of an isomerase, an isomerase having a mutated active site, or the crystal of some other protein with significant sequence and/or structural homology to an isomerase may be determined using the coordinates given in Table 1. This method provides sufficient structural form for the unknown crystal more efficiently than attempting to determine such information ab initio. In addition, this method can be used to determine whether or not a given isomerase in question falls within the scope of this invention.
As further disclosed herein, isomerases and mutants thereof may be crystallized in the presence or absence of substrates and substrate analogs. The crystal structures of a series of complexes may then be solved by molecular replacement and compared to that of the wild-type isomerase to assist in determination of suitable replacements for R-groups within the active site, thus making isomerase mutants according to the present invention.
All mutants of the present inventions may be modeled using the information disclosed herein without necessarily having to crystallize and solve the structure for each and every mutant. For example, one skilled in the art may use one of several specialized computer programs to assist in the process of designing isomerases having mutated active sites relative to the wild-type isomerase. Examples of such programs include: GRID (Goodford, 1985, J. Mod. Chem., 28:849-857), MCSS (Miranker and Karplus, 1991, Proteins: Structure, Function and Genetics, 11:29-34); AUTODOCK (Goodsell and Olsen, 1990, Proteins. Structure, Fumtion, and Genetics, 8:195-202); and DOCK (Kuntz et al., 1982, J. Mot BioL, 161:269-288), and the like, as well as those discussed in the Examples below. In addition, specific computer programs are also available to evaluate specific substrate-active site interactions and the deformation energies and electrostatic interactions resulting therefrom. MODELLER is a computer program often used for homology or comparative modeling of the three-dimensional structure of a protein. A. Saii & T. L. Blundell. J. Mol. Biol. 234:779-815, 1993. A sequence to be modeled is aligned with one or more known related structures and the MODELLER program is used to calculate a full-atom model, based on optimum satisfaction of spatial restraints. Such restraints can include, inter alia, homologous structures, site-directed mutagenesis, fluorescence spectroscopy, NMR experiments, or atom-atom potentials of mean force.
The present invention enables isomerase mutants to be made and the crystal structure thereof to be solved. Moreover, by virtue of the present invention, the location of the active site and the interface of substrate therewith permit the identification of desirable R-groups for introduction by mutagenesis.
The three-dimensional coordinates of the isomerases provided herein may additionally be used to predict the activity and or substrate specificity of a protein whose primary amino acid sequence suggests that it may have isomerase activity. The family of CHI-related enzymes is defined, in part, by a number of conserved amino acid residues including, for example, residues spanning β3a, β3b, α4, and α6 in the three-dimensional structure. By employing the three-dimensional coordinates disclosed herein and computer modeling programs, structural comparisons of CHI can be made with a putative enzyme. Differences between the two would provide the skilled artisan with information regarding the activity and/or substrate specificity of the putative enzyme.
Thus, in another embodiment of the invention, there is provided a method of predicting the activity and/or substrate specificity of an isomerase or putative isomerase comprising (a) generating a three-dimensional representation of a known isomerase (e.g., chalcone isomerase) using three-dimensional coordinate data, (b) generating a predicted three-dimensional representation of a putative isomerase, and (c) comparing the representation of the known isomerase with the representation of the putative isomerase, wherein the differences between the two representations are predictive of activity and/or substrate specificity of the putative isomerase.
In a further embodiment of the present invention, there is also provided a method of identifying a potential substrate of an isomerase comprising (a) defining the active site of an isomerase (e.g., chalcone isomerase) based on the atomic coordinates of the isomerase, (b) identifying a potential substrate that fits the defined active site, and (c) contacting the isomerase with the potential substrate of (b) and determining the activity thereon. Techniques for computer modeling and structural comparisons similar to those described herein for predicting putative isomerase activity and/or substrate specificity can be used to identify novel substrates for isomerases. The plurality of atomic coordinates that can be used to define the active site of an isomerase include those set forth in PDB Accession Nos: 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), 1JEP (SEQ ID NO:9) and Table 1 (SEQ ID NOs:9-11). A subset or portion of these atomic coordinates can also be used, for example, those atomic coordinates defining the amino acid residues which comprise the enzymatic active site.
In addition, the structural coordinates and three-dimensional models disclosed herein can be used to design or identify isomerase inhibitors. Using the modeling techniques disclosed herein, potential inhibitor structures can be modeled with the isomerase active site and those that appear to interact therewith can subsequently be tested in activity assays in the presence of substrate.
Methods of using crystal structure data to design binding agents or substrates are known in the art. Thus, the crystal structure data provided herein can be used in the design of new or improved inhibitors, substrates or binding agents. For example, the isomerase polypeptide coordinates can be superimposed onto other available coordinates of similar enzymes to identify modifications in the active sites of the enzymes to create novel by-products of enzymatic activity or to modulate flavonoid synthesis. Alternatively, the isomerase polypeptide coordinates can be superimposed onto other available coordinates of similar enzymes which have substrates or inhibitors bound to them to give an approximation of the way these and related substrates or inhibitors might bind to an isomerase. Alternatively, computer programs employed in the practice of rational drug design can be used to identify compounds that reproduce interaction characteristics similar to those found between a isomerase polypeptide and a co-crystallized substrate. Furthermore, detailed knowledge of the nature of binding site interactions allows for the modification of compounds to alter or improve solubility, pharmacokinetics, etc. without affecting binding activity.
Computer programs are widely available that are capable of carrying out the activities necessary to design agents using the crystal structure information provided herein. Examples include, but are not limited to, the computer programs listed below:

    
    
        CATALYST DATABASESTM—an information retrieval program accessing chemical databases such as BioByte Master File, Derwent WDI and ACD;
        CATALYST/HYPOTM—generates models of compounds and hypotheses to explain variations of activity with the structure of drug candidates;
        LUDITM—fits molecules into the active site of a protein by identifying and matching complementary polar and hydrophobic groups;
        LEAPFROGTM—“grows” new ligands using a genetic algorithm with parameters under the control of the user.
    
    


In addition, various general purpose machines may be used with programs written in accordance with the teachings herein, or it may be more convenient to construct more specialized apparatus to perform the operations. However, preferably the embodiment is implemented in one or more computer programs executing on programmable systems each comprising at least one processor, at least one data storage system (including volatile and non-volatile memory and/or storage elements), at least one input device, and at least one output device. The program is executed on the processor to perform the functions described herein.
Each such program may be implemented in any desired computer language (including machine, assembly, high level procedural, object oriented programming languages, or the like) to communicate with a computer system. In any case, the language may be a compiled or interpreted language. The computer program will typically be stored on a storage media or device (e.g., ROM, CD-ROM, or magnetic or optical media) readable by a general or special purpose programmable computer, for configuring and operating the computer when the storage media or device is read by the computer to perform the procedures described herein. The system may also be considered to be implemented as a computer-readable storage medium, configured with a computer program, where the storage medium so configured causes a computer to operate in a specific and predefined manner to perform the functions described herein.
Embodiments of the invention include systems (e.g., interne based systems), particularly computer systems which store and manipulate the coordinate and sequence information described herein. One example of a computer system 100 is illustrated in block diagram form in FIG. 5. As used herein, “a computer system” refers to the hardware components, software components, and data storage components used to analyze the coordinates and sequences as set forth in Accession Nos. 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), and Table 1 (SEQ ID NOs:9-11). The computer system 100 typically includes a processor for processing, accessing and manipulating the sequence data. The processor 105 can be any well-known type of central processing unit, such as, for example, the Pentium III from Intel Corporation, or similar processor from Sun, Motorola, Compaq, AMD or International Business Machines.
Typically the computer system 100 is a general purpose system that comprises the processor 105 and one or more internal data storage components 110 for storing data, and one or more data retrieving devices for retrieving the data stored on the data storage components. A skilled artisan can readily appreciate that any one of the currently available computer systems are suitable.
In one particular embodiment, the computer system 100 includes a processor 105 connected to a bus which is connected to a main memory 115 (preferably implemented as RAM) and one or more internal data storage devices 110, such as a hard drive and/or other computer readable media having data recorded thereon. In some embodiments, the computer system 100 further includes one or more data retrieving device 118 for reading the data stored on the internal data storage devices 110.
The data retrieving device 118 may represent, for example, a floppy disk drive, a compact disk drive, a magnetic tape drive, or a modem capable of connection to a remote data storage system (e.g., via the internet) etc. In some embodiments, the internal data storage device 110 is a removable computer readable medium such as a floppy disk, a compact disk, a magnetic tape, etc. containing control logic and/or data recorded thereon. The computer system 100 may advantageously include or be programmed by appropriate software for reading the control logic and/or the data from the data storage component once inserted in the data retrieving device.
The computer system 100 includes a display 120 which is used to display output to a computer user. It should also be noted that the computer system 100 can be linked to other computer systems 125a-c in a network or wide area network to provide centralized access to the computer system 100.
Software for accessing and processing the coordinate and sequences described herein, (such as search tools, compare tools, and modeling tools etc.) may reside in main memory 115 during execution.
For the first time, the present invention permits the use of molecular design techniques to design, select and synthesize novel enzymes, chemical entities and compounds, including inhibitory compounds, capable of binding to an isomerase polypeptide (e.g., a chalcone isomerase polypeptide), in whole or in part.
One approach enabled by this invention, is to use the structural coordinates as set forth in Accession Nos. 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), 1JEP (SEQ ID NO:9), and Table 1 (SEQ ID NOs:9-11) to design new enzymes capable of synthesizing novel flavonoids. For example, isomerases generate molecular diversity in their products by utilizing different starter molecules. The structural coordinates disclosed herein allow the elucidation of the nature by which isomerases achieve starter molecule selectivity and control flavonoid diversity and synthesis. Accordingly, the invention allows for the strategic development and biosynthesis of more diverse flavonoids and demonstrates a structural basis for control of flavonoid synthesis. In addition, the structural coordinates allow for the development of substrates or binding agents that bind to the polypeptide and alter the physical properties of the compounds in different ways, e.g., solubility.
In another approach an isomerase polypeptide crystal is probed with molecules composed of a variety of different chemical entities to determine optimal sites for interaction between candidate binding molecules (e.g., substrates) and the isomerase (e.g., chalcone isomerase).
In another embodiment, an approach made possible and enabled by this invention, is to screen computationally small molecule data bases for chemical entities or compounds that can bind in whole, or in part, to an isomerase polypeptide or fragment thereof. In this screening, the quality of fit of such entities or compounds to the binding site may be judged either by shape complementarity or by estimated interaction energy. Meng, E. C. et al., J. Comp. Chem, 13, pp. 505-524 (1992).
Chalcone isomerase is one member of a family of isomerase polypeptides, many of which have similar functional activity. In addition, many isomerase polypeptides may crystallize in more than one crystal form. Accordingly, the structural coordinates of chalcone isomerase, or portions thereof, as provided by this invention are particularly useful to solve the structure, function or activity of other crystal forms of isomerase polypeptides. They may also be used to solve the structure of an isomerase or a chalcone isomerase mutant.
One method that may be employed for this purpose is molecular replacement. In this method, the unknown crystal structure, whether it is another isomerase crystal form, chalcone isomerase, chalcone isomerase mutant, an isomerase complexed with a substrate or other molecule, or the crystal of some other protein with significant amino acid sequence homology to any isomerase polypeptide, may be determined using the structure coordinates as provided in Accession Nos. 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), 1JEP (SEQ ID NO:9) and Table 1 (SEQ ID NOs:9-11). This method will provide an accurate structural form for the unknown crystal more quickly and efficiently than attempting to determine such information ab initio.
In addition, in accordance with the present invention, an isomerase, chalcone isomerase or chalcone isomerase mutant may be crystallized in association or complex with known isomerase binding agents, substrates, or inhibitors. The crystal structures of a series of such complexes may then be solved by molecular replacement and compared with that of wild-type isomerase polypeptides. Potential sites for modification within the isomerase polypeptide may thus be identified. This information provides an additional tool for determining the most efficient binding interactions between an isomerase and a chemical entity, substrate or compound.
All of the complexes referred to above may be studied using well-known X-ray diffraction techniques and may be refined to 2-3 Å resolution X-ray data to an R value of about 0.20 or less using computer software, such as X-PLOR (ale University, 1992, distributed by Molecular Simulations, Inc.). See, e.g., Blundel & Johnson, supra; Methods in Enzymology, vol. 114 and 115, H. W. Wyckoff et al., eds., Academic Press (1985). This information may thus be used to optimize known classes of isomerase substrates or binding agents (e.g., inhibitors), and to design and synthesize novel classes of isomerases, substrates, and binding agents (e.g., inhibitors).
The design of substrates, compounds or binding agents that bind to or inhibit a chalcone isomerase polypeptide according to the invention generally involves consideration of two factors. First, the substrate, compound or binding agent must be capable of physically and structurally associating with the isomerase polypeptide. Non-covalent molecular interactions important in the association of a polyketide isomerase with a substrate include hydrogen bonding, van der Waals and hydrophobic interactions, and the like.
Second, the substrate, compound or binding agent must be able to assume a conformation that allows it to associate with an isomerase polypeptide. Although certain portions of the substrate, compound or binding agent will not directly participate in this association, those portions may still influence the overall conformation of the molecule. This, in turn, may have a significant impact on 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 binding site, e.g., active site or accessory binding site of a polyketide isomerase (e.g., a chalcone isomerase polypeptide), or the spacing between functional groups of a substrate or compound comprising several chemical entities that directly interact with an isomerase.
The potential binding effect of a substrate or chemical compound on an isomerase or the activity of a newly synthesized or mutated isomerase might have on a known substrate may be analyzed prior to its actual synthesis and testing by the use of computer modeling techniques. For example, if the theoretical structure of the given substrate or compound suggests insufficient interaction and association between it and an isomerase, synthesis and testing of the compound may not be warranted. However, if computer modeling indicates a strong interaction, the molecule may then be tested for its ability to bind to, initiate catalysis or elongation of a flavonoid by an isomerase. Methods of assaying for isomerase activity are known in the art (as identified and discussed herein). Methods for assaying the effect of a newly created isomerase or a potential substrate or binding agent can be performed in the presence of a known binding agent or isomerase. For example, the effect of the potential binding agent can be assayed by measuring the ability of the potential binding agent to compete with a known substrate.
A mutagenized isomerase, novel isomerase, substrate or other binding compound of an isomerase may be computationally evaluated and designed by means of a series of steps in which chemical entities or fragments are screened and selected for their ability to associate with the individual binding pockets or other areas of the isomerase.
One skilled in the art may use one of several methods to screen chemical entities or fragments for their ability to associate with an isomerase and more particularly with the individual binding pockets of a chalcone isomerase polypeptide. This process may begin by visual inspection of, for example, the active site on the computer screen based on the coordinates in Accession Nos. 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), and Table 1 (SEQ ID NOs:9-11). Selected fragments or substrates or chemical entities may then be positioned in a variety of orientations, or docked, within an individual binding pocket of an isomerase. Docking may be accomplished using software such as QUANTA and SYBYL, followed by energy minimization and molecular dynamics with standard molecular mechanics forcefields, such as CHARMM and AMBER.
Specialized computer programs may also assist in the process of selecting fragments or chemical entities. These include:
1. GRID (Goodford, P. J., “A Computational Procedure for Determining Energetically Favorable Binding Sites on Biologically Important Macromolecules”, J. Med. Chem., 28, pp. 849-857 (1985)). GRID is available from Oxford University, Oxford, UK.
2. MCSS (Miranker, A. and M. Karplus, “Functionality Maps of Binding Sites: A Multiple Copy Simultaneous Search Method.” Proteins: Structure. Function and Genetics, 11, pp. 29-34 (1991)). MCSS is available from Molecular Simulations, Burlington, Mass.
3. AUTODOCK (Goodsell, D. S. and A. J. Olsen, “Automated Docking of Substrates to Proteins by Simulated Annealing”, Proteins: Structure. Function, and Genetics, 8, pp. 195-202 (1990)). AUTODOCK is available from Scripps Research Institute, La Jolla, Calif.
4. DOCK (Kuntz, I. D. et al., “A Geometric Approach to Macromolecule-Ligand Interactions”, J. Mol. Biol., 161, pp. 269-288 (1982)). DOCK is available from University of California, San Francisco, Calif.
Once suitable substrates, chemical entities or fragments have been selected, they can be assembled into a single polypeptide, compound or binding agent (e.g., an inhibitor). Assembly may be performed by visual inspection of the relationship of the fragments to each other on the three-dimensional image displayed on a computer screen in relation to the structure coordinates of the molecules as set forth in Accession Nos. 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), and Table 1 (SEQ ID NOs:9-11). This would be followed by manual model building using software such as QUANTA or SYBYL.
Useful programs to aid one of skill in the art in connecting the individual chemical entities or fragments include:
1. CAVEAT (Bartlett, P. A. et al, “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, pp. 182-196 (1989)). CAVEAT is available from the University of California, Berkeley, Calif.
2. 3D Database systems such as MACCS-3D (MDL Information Systems, San Leandro, Calif.). This area is reviewed in Martin, Y. C., “3D Database Searching in Drug Design”, J. Med. Chem., 35, pp. 2145-2154 (1992)).
3. HOOK (available from Molecular Simulations, Burlington, Mass.).
In addition to the method of building or identifying novel enzymes or an isomerase substrate or binding agent in a step-wise fashion one fragment or chemical entity at a time as described above, substrates, inhibitors or other isomerase interactions may be designed as a whole or “de novo” using either an empty active site or optionally including some portion(s) of known substrates, binding agents or inhibitors. These methods include:
1. LUDI (Bohm, H.-J., “The Computer Program LUDI: A New Method for the De Novo Design of Enzyme Inhibitors”, J. Comp. Aid. Molec. Design, 6, pp. 61-78 (1992)). LUDI is available from Biosym Technologies, San Diego, Calif.
2. LEGEND (Nishibata, Y. and A. Itai, Tetrahedron, 47, p. 8985 (1991)). LEGEND is available from Molecular Simulations, Burlington, Mass.
3. LEAPFROG (available from Tripos Associates, St. Louis, Mo.).
Other molecular modeling techniques may also be employed in accordance with this invention. See, e.g., Cohen, N. C. et al., “Molecular Modeling Software and Methods for Medicinal Chemistry”, J. Med. Chem., 33, pp. 883-894 (1990). See also, Navia, M. A. and M. A. Murcko, “The Use of Structural Information in Drug Design”, Current Opinions in Structural Biology, 2, pp. 202-210 (1992).
Once a substrate, compound or binding agent has been designed or selected by the above methods, the efficiency with which that substrate, compound or binding agent may bind to an isomerase may be tested and optimized by computational evaluation.
A substrate or compound designed or selected as an isomerase binding agent may be further computationally optimized so that in its bound state it would preferably lack repulsive electrostatic interaction with the target site. 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 binding agent and the isomerase when the binding agent is bound to the isomerase, preferably make a neutral or favorable contribution to the enthalpy of binding.
Specific computer software is available in the art to evaluate compound deformation energy and electrostatic interaction. Examples of programs designed for such uses include: GAUSSIAN 92, revision C (M. J. Frisch, Gaussian, Inc., Pittsburgh, Pa., 1992); AMBER, version 4.0 (P. A. Kollman, University of California at San Francisco, 1994); QUANTA/CHARMM (Molecular Simulations, Inc., Burlington, Mass. 1994); and INSIGHT II/DISCOVER (Biosysm Technologies Inc., San Diego, Calif., 1994). These programs may be implemented, for example, using a Silicon Graphics workstation, IRIS 4D/35 or IBM RISC/6000 workstation model 550. Other hardware systems and software packages will be known to those skilled in the art of which the speed and capacity are continually modified.
Once an isomerase, isomerase substrate or isomerase binding agent has been 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, e.g., the replacement group will have approximately the same size, shape, hydrophobicity and charge as the original group. Such substituted chemical compounds may then be analyzed for efficiency of fit to a polyketide isomerase substrate or fit of a modified substrate to an isomerase having a structure defined by the coordinates in Accession Nos. 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), 1JEP (SEQ ID NO:9) and Table 1 (SEQ ID NOs:9-11), by the same computer methods described, above.
Conserved regions of the flavonoid family isomerases lend themselves to the methods and compositions of the invention. For example, a number of isomerases have conserved residues present within their active sites (as described more fully below). Accordingly, modification to the active site of chalcone isomerase or a chalcone isomerase substrate can be extrapolated to other conserved members of the family of isomerases.
Functional fragments of isomerase polypeptides such as, for example, fragments of chalcone isomerase, can be designed based on the crystal structure and atomic coordinates described herein. Fragments of a chalcone isomerase polypeptide and the fragment's corresponding atomic coordinates can be used in the modeling described herein. In addition, such fragments may be used to design novel substrates or modified active sites to create new diverse flavonoids.
In one embodiment of the present invention, the crystal structure and atomic coordinates allow for the design of novel isomerases and novel isomerase substrates. The development of new isomerases will lead to the development a biodiverse library of flavonoids for use as therapeutics (e.g., as antibiotics, anti-cancer agents, anti-fungal agents) as described herein or known in the art. In vitro assay systems for production and determination of activity are known in the art. For example, antibiotic activities of novel products of the polyketide pathway and flavonoid pathway can be measured by any number of anti-microbial techniques currently used in hospitals and laboratories. In addition, anticancer activity can be determined by contacting cells having a cell proliferative disorder with a newly synthesized flavonoid and measuring the proliferation or apoptosis of the cells before and after contact with the flavonoid. Specific examples of apoptosis assays are provided in the following references: Lymphocyte: C. J. Li et al., Science, 268:429-431, 1995; D. Gibellini et al., Br. J. Haematol. 89:24-33, 1995; S. J. Martin et al., J. Immunol. 152:330-42, 1994; C. Terai et al., J. Clin Invest. 87:1710-5, 1991; J. Dhein et al., Nature 373:438-441, 1995; P. D. Katsikis et al., J. Exp. Med. 1815:2029-2036, 1995; Michael O. Westendorp et al., Nature 375:497, 1995; DeRossi et al., Virology 198:234-44, 1994. Fibroblasts: H. Vossbeck et al., Int. J. Cancer 61:92-97, 1995; S. Goruppi et al., Oncogene 9:1537-44, 1994; A. Fernandez et al., Oncogene 9:2009-17, 1994; E. A. Harrington et al., Embo J. 13:3286-3295, 1994; N. Itoh et al., J. Biol. Chem. 268:10932-7, 1993. Neuronal Cells: G. Melino et al., Mol. Cell. Biol. 14:6584-6596, 1994; D. M. Rosenbaum et al., Ann. Neurol. 36:864-870, 1994; N. Sato et al., J. Neurobiol 25:1227-1234, 1994; G. Ferrari et al., J. Neurosci. 1516:2857-2866, 1995; A. K. Talley et al., Mol. Cell Biol. 1585:2359-2366, 1995; A. K. Talley et al., Mol. and Cell. Biol. 15:2359-2366, 1995; G. Walkinshaw et al., J. Clin. Invest. 95:2458-2464, 1995. Insect Cells: R. J. Clem et al., Science 254:1388-90, 1991; N. E. Crook et al., J. Virol. 67:2168-74, 1993; S. Rabizadeh et al., J. Neurochem. 61:2318-21, 1993; M. J. Birnbaum et al., J. Virol 68:2521-8, 1994; R. J. Clem et al., Mol. Cell. Biol. 14:5212-5222, (1994). Other assays are well within the ability of those of skill in the art.
Production of novel flavonoids or isomerases can be carried out in culture. For example, mammalian expression constructs carrying isomerases can be introduced into various cell lines such as CHO, 3T3, HL60, Rat-1, or Jurkart cells, for example. In addition, SF21 insect cells may be used in which case the isomerase gene is expressed using an insect heat shock promoter.
In another embodiment of the present invention, once a novel substrate or binding agent is developed by the computer methodology discussed above, the invention provides a method for determining the ability of the substrate or agent to be acted upon by an isomerase. The method includes contacting components comprising the substrate or agent and an isomerase, or a recombinant cell expressing an isomerase, under conditions sufficient to allow the substrate or agent to interact and determining the affect of the agent on the activity of the polypeptide. The term “affect”, as used herein, encompasses any means by which protein activity can be modulated, and includes measuring the interaction of the agent with the isomerase polypeptide by physical means including, for example, fluorescence detection of the binding of an agent to the polypeptide. Such agents can include, for example, polypeptides, peptidomimetics, chemical compounds, small molecules, substrates and biologic agents as described herein. Examples of small molecules include but are not limited to small peptides or peptide-like molecules.
Contacting or incubating includes conditions which allow contact between the test agent or substrate and an isomerase or modified isomerase polypeptide or a cell expressing an isomerase or modified isomerase polypeptide. Contacting includes in solution and in solid phase. The substrate or test agent may optionally be a combinatorial library for screening a plurality of substrates or test agents. Agents identified in the method of the invention can be further evaluated by chromatography, cloning, sequencing, and the like.
In yet another embodiment, the present invention provides a computer for producing a three-dimensional representation of a molecule or molecular complex or a homologue of said molecule or molecular complex, wherein said molecule or molecular complex or a homologue of said molecule or molecular complex comprises an active site defined by atomic coordinates are as set forth in PDB Accession Nos: 1EYP (SEQ ID NOs:9-11), 1EYQ (SEQ ID NO:9), 1FM7 (SEQ ID NO:9), 1FM8 (SEQ ID NO:9), 1JEP (SEQ ID NO:9), or Table 1 (SEQ ID NOs:9-11), wherein said computer comprises:

    
    
        i) a computer-readable data storage medium comprising a data storage material encoded with computer-readable data, wherein said data comprises atomic coordinates are as set forth in PDB Accession Nos: 1EYP, 1EYQ, 1FM7, 1FM8, 1JEP, or Table 1;
        (ii) a working memory for storing instructions for processing said computer-readable data;
        (iii) a central-processing unit coupled to said working memory and to said computer-readable data storage medium for processing said computer-machine readable data into said three-dimensional representation; and
        (iv) a display coupled to said central-processing unit for displaying said three-dimensional representation.
    
    


Such a computer could also be used to determine at least a portion of the atomic coordinates corresponding to X-ray diffraction data obtained from a molecule or molecular complex or a homologue of said molecule or molecular complex.
In yet another embodiment, the present invention provides methods of screening compounds to determine whether they are isomerase substrates, said method comprising:

    
    
        a) determining the points of interaction between an isomerase and a substrate or product therefor;
        b) selecting compound(s) having similar interaction with said isomerase; and
        c) testing the selected compound for the ability to be converted by said isomerase.
    
    


An alternative method of screening compounds to determine whether they are isomerase substrates comprises:

    
    
        a) selecting compound(s) having points of interaction with said isomerase, wherein similar points of interaction have been determined between said isomerase and a substrate or product therefor; and
        b) testing the selected compound for the ability to be converted by said isomerase.
    
    


Another alternative method of screening compounds to determine whether they are isomerase substrates comprises:

    
    
        testing a compound for the ability to be converted by said isomerase,
        wherein said compound has been selected as having points of interaction with said isomerase, and
        wherein similar points of interaction have been determined between said isomerase and a substrate or product therefor.
    
    


In another embodiment, the present invention provides methods for screening for compounds that inhibit an isomerase comprising:

    
    
        a) determining the points of interaction between an isomerase and a substrate or product therefor;
        b) selecting compound(s) having similar interaction with said isomerase; and
        c) testing the selected compound for the ability to inhibit the activity of said isomerase.
    
    


An alternative method for screening for compounds that inhibit an isomerase comprises:

    
    
        a) selecting compound(s) having points of interaction with an isomerase, wherein similar points of interaction have been determined between said isomerase and a substrate or product therefor; and
        b) testing the selected compound for the ability to inhibit the activity of said isomerase.
    
    


Another alternative method for screening for compounds that inhibit an isomerase comprises:

    
    
        testing a compound for the ability to inhibit the activity of said isomerase,
        wherein said compound has been selected as having points of interaction with said isomerase, and
        wherein similar points of interaction have been determined between said isomerase and a substrate or product therefor.
    
    


The present invention also claims a compound identified by these methods and a composition comprising such a compound and an acceptable carrier therefor.
Although methods and materials similar or equivalent to those described herein can be used to practice the invention, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The invention will now be described in greater detail by reference to the following non-limiting examples.
EXAMPLES
Expression, Mutagenesis, and Purification
Alfalfa CHI (SEQ ID NO:1) cDNA was PCR amplified and inserted into the pHIS8 expression vector (Jez et al., Biochemistry 39:890-902, 2000). The CHI Y106F mutant (SEQ ID NO:13) was generated with the QuikChange (Stratagene) PCR method. N-terminal His-tagged protein was expressed in E. coli BL21(DE3) cells. Tagged CHI was purified from sonicates using a Ni2+-NTA (Qiagen) column. Thrombin digestion removed the histidine tag, and the protein was passed over a Ni2+-NTA column. Digested CHI was depleted of thrombin using a benzamidine-Sepharose column. Gel filtration on a SUPERDEX-75 (Pharmacia) FPLC column was the final purification step.
Enzyme Assays
CHI assays were performed at 25° C. in a 0.5 ml reaction volume containing 0.05 M Hepes (pH 7.5), 50 μM 6′-deoxychalcone, and 3% ethanol as co-solvent. Time-dependent decreases in 6′-deoxychalcone absorbance (λmax=390 nm; ε=29,400 M−1 cm−1) were monitored with a Beckman DU-640 spectrophotometer. Determination of steady-state kinetic constants used the standard assay system with varied concentrations of substrate (2-50 μM) following fitting to the Michaelis-Menton equation using KALEIDAGRAPH (Abelbeck Software).
Crystallization, Structure Determination, and Refinement of the Native Structure
Crystals of CHI (SEQ ID NO:1) were grown at 4° C. by vapor diffusion using the hanging drop method. A 2 μl drop containing a 1:1 mixture of 25 mg ml−1 CHI and crystallization buffer (25% glycerol, 1.8-2.0 M ammonium sulfate and 0.05 M PIPES, pH 6.5) yielded diffraction quality crystals within a few days at 4° C. Crystals grew in space group P6522 with unit cell dimensions of a=90.37 Å; c=352.86 Å with two molecules per asymmetric unit and a solvent content of 72%. Native CHI diffraction data (105 K) were collected at beamline 7-1 of the Stanford Synchrotron Radiation Source (SSRL 7-1) on a 30 cm MAR imaging plate system. For generation of heavy atom derivatives, CHI crystals were soaked in mother liquor with either 1.2 mM K2OsCl6 or 1 mM HgCl2 for 12-16 hours. Heavy atom data (105 K) were collected at SSRL 9-1 on a 30 cm MAR imaging plate system. All images were indexed and integrated using DENZO and the reflections merged with SCALEPACK (Otwinowski, Z. & Minor, W., Methods Enzymol. 276:307-326, 1997). Data reduction was completed using programs from CCP4 (Collaborative Computational Project 4 (CCP4) Acta Crystallogr. D53:240-255, 1994) (See Table 3). Heavy atom sites were located with SOLVE (Terwilliger, T. C. & Berendzen, J. Acta Crystallogr. D55:849-861, 1999) Refinement of sites and location of additional sites used MLPHARE (Otwinowski, Z. ML-PHARE in Daresbary Study Weekend Proceedings (CCP4, SERC Daresbary Laboratory, Warrington, UK; 1991). SHARP was used for phase calculation and heavy atom refinement (de La Fortelle, E. & Bricogne, G. Methods Enzymol. 276:472-494, 1997). This set of experimental phases was improved and extended using solvent flipping with SOLOMON (Abrahams, J. P. & Leslie, A. G. W. Acta Crystallogr. D52:30-42, 1996). Model building was performed with 0. CNS was used for refinement (Brünger, A. T. et al. Acta Crystallogr. D54:905-921, 1998). The initial model was subjected to simulated annealing, positional refinement, and group B-factor refinement with strict non-crystallographic symmetry maintained between both molecules in the asymmetric unit. In subsequent rounds of model building and refinement, non-crystallographic constraints were released and water molecules were added using CNS to yield the R-factors shown in Table 3. The final model included residues 4 to 215 of monomer A (SEQ ID NO:9), residues 3 to 38 (SEQ ID NO:10) and 45 to 215 (SEQ ID NO:11) of monomer B. The quality of the CHI model was checked with PROCHECK (Laskowski, R. A., MacArthur, M. W., Moss, D. S., & Thornton, J. M. J. Appl. Crystallogr. 26:283-291, 1993). A total of 89.6% of the residues in CHI are in the most favored regions of the Ramachandran plot and 10.4% are in the additional allowed region.
Overall Structure
Expression of alfalfa CHI (SEQ ID NO:1) in E. coli yielded active enzyme that was purified and crystallized. The overall structure of CHI resembles an upside-down bouquet that adopts an open-faced β-sandwich fold (FIGS. 1B and 1C). A large β-sheet β3a-β3f) and a layer of α-helices (α1-α7) comprise the core structure with three short β-strands (β1a, β1b, β2) on the opposite side of the large β-sheet. A search of the Protein Data Bank using DALI revealed no other structurally homologous folds. In addition, a PSI-BLAST search of sequence databases showed that CHI-like sequences are currently found typically in plants and these sequences display no detectable homology with other proteins. These results imply that the CHI three-dimensional fold and enzymatic activity are typically found in the plant kingdom. Amino acid sequence comparison of CHIs from a variety of advanced land plants reveals high homology (49% to 82% amino acid sequence identity) with regions of conservation spread uniformly throughout the primary structure (FIG. 1D, SEQ ID NOs:1-8). Interestingly, there is conservation of residues spanning β3a, β3b, α4, and α6 in the three-dimensional structure among CHIs from different species. Notably, these structural elements form the active site on the protein surface.
Accumulating data suggests that co-localization of proteins in loosely associated macromolecular complexes is a fundamental component of cellular processes, including flavonoid biosynthesis. CHI and other flavonoid biosynthetic enzymes may associate to provide efficient channeling of substrates and products as shown recently in Arabidopsis thaliana. Although the three short β-strands (β1a, β1b, β2) on the backside of the CHI structure form a relatively flat surface that would be ideal for protein-protein interactions, both gel filtration and analytical ultracentrifugation experiments failed to detect association of alfalfa CHI (SEQ ID NO:1) and alfalfa chalcone synthase 2 in vitro.







TABLE 3







Crystallographic data, phasing, and refinement information












Native
K2OsCl6
HgCl2
Naringenin














Wavelength (Å)
1.08
0.98
0.98
0.95


Resolution range (Å)
38.1-2.50
78.0-3.24
76.0-3.26
46.7-1.85


Observations
86,781
44,070
39,193
697,121


Unique reflections
27,863
24,966
22,238
70,889


Completeness1 (%)
90.7 (61.7)
99.2 (99.6)
95.8 (98.2)
85.5 (60.6)


I/□1
19.4 (2.0) 
9.3 (2.6)
17.5 (7.4) 
28.0 (2.0) 


Rsym1,2 (%)
 5.4 (36.5)
10.3 (31.2)
 5.8 (12.0)
 4.8 (45.4)


POP3 (acentric/centric)

2.55/1.85
1.58/1.07


Rcullis4 (iso/ano)

0.52/0.76
0.72/0.81


Rcryst5/Rfree6 (%)
24.9/28.0


23.7/26.2


Protein atoms
3181


3222


Water molecules
94


382


Ligand atoms7



40 nar/20 sul


R.m.s.d. bonds (Å)
0.019


0.019


R.m.s.d. angles (°)
2.0


2.1


average B-factor - protein (Å2)
60.6


46.8


average B-factor - water (Å2)
62.7


50.1





1Number in parenthesis is for highest resolution shell;


2Rsym = Σ|Ih − <Ih>|/ ΣIh, where <Ih>, is the average intensity over symmetry equivalent reflections;


3Power of Phasing = <|FH(calc)/|E|>, where FH(calc)is the calculated difference and E is the lack of closure;


4Rcullis = Σ|E|/Σ|FPH − FP|;


5R-factor = Σ|Fobs − Fcalc|/ΣFobs, where summation is over the data used for refinement;


6Rfree-factor was calculated using 5% of data excluded from refinement;


7nar = naringenin and sul = sulfate.






Crystallization, Structure Determination, and Refinement of the CHI•Naringenin Complex Structure

Crystals of the CHI•naringenin (SEQ ID NO:5) complex (P6522; a=89.47 Å; c=351.19 Å) (and other co-complexes) were grown as above from a crystallization buffer containing 2.5 mM (2S/2R)-naringenin and 5% ethanol. Data (105 K) were collected at SSRL 9-2 with a Quantum 4 CCD detector. Images were processed as above. Following rigid-body refinement with CNS, electron density resembling naringenin was observed in each monomer and modeled as such. In subsequent rounds of refinement and rebuilding, the R-factors converged to those listed in Table 3. The final model includes residues 4 to 215 of both monomers.
(2S)-Naringenin Binding and Reaction Stereoselectivity
The location of (2S)-naringenin in the CHI (SEQ ID NO:5) structure defines the active site (FIG. 2). Although a commercially obtained mixture of (2S)- and (2R)-naringenin was used for co-crystallization, only the (2S)-isomer bound the CHI active site. The position of the (2S)-naringenin binding cleft is consistent with inactivation studies that suggested a cysteine residue (Cys 114 in alfalfa CHI, SEQ ID NO:1) is proximal to the active site. In the CHI structure, Cys 114 is near the binding cleft but does not directly contact (2S)-naringenin. The active site cleft is largely apolar and consists of residues from β3a (Arg 36, Gly 37, Leu 38), β3b (Phe 47, Thr 48, Ile 50), α4 (Tyr 106, Lys 109, Val 110, Asn 113), and α6 (Thr 190, Met 191) (FIG. 2B) (SEQ ID NO:9). The apolar methylene carbons of Arg 36 (SEQ ID NO:9) are positioned by a restraining charge-charge interaction from the 8-guanido group to Glu 200. In addition, the methylene carbons of Lys 109 (SEQ ID NO:9) are fixed by a charge-charge interaction between the side chain amine and Glu 112. Except for Thr 190 and Met 191, the residues contacting (2S)-naringenin are identical among CHIs from different plants (FIG. 1D) (SEQ ID NOs:1-8). Although van der Waals contacts dominate the interactions between CHI and (2S)-naringenin, two hydrogen bond interactions exist. The first is mediated by the side chain hydroxyl moiety of Thr 48 bound to the 4′-hydroxyl group of (2S)-naringenin (SEQ ID NO:5). The second interaction is between a water molecule and the ligand ketone (FIG. 2B). This water molecule and its connected network of hydrogen bonds occupy the same position in the apoenzyme structure. The overall surface topology of the cleft tightly sequesters the (2S)-naringenin molecule (FIG. 2C). The CHI•naringenin complex (SEQ ID NO:9) explains the stereochemical preference of the cyclization reaction; moreover, it suggests why CHIs from different species show moderate selectivity for chalcone and 6′-deoxychalcone as substrates.
Modeling of chalcone, based on the position of (2S)-naringenin, shows that a slight rotation of the trihydroxyl-ring outward in the direction of the active site opening places the 2′-hydroxyl group in position for nucleophilic attack on the α,β-unsaturated double bond of the coumaroyl moiety (FIG. 3A). This rotation preserves the position of the chalcone backbone and the hydrogen bonds between Thr 190 and the water molecule at the backside of the binding site. Formation of (2R)-naringenin would require substantial rearrangements in the active site of CHI due to significant steric clashes between the trihydroxyl-ring and CHI side chains. Although rotation of the trihydroxyl-ring away from the active site entrance could reposition the 2′-hydroxyl group for attack on the opposite face of the α,β-double bond, the side chain of Val 110 sterically prevents this movement from occurring (FIG. 3B). In addition, the opposite side of the substrate double bond could be positioned for attack by the 2′-hydroxyl group in the formation of (2R)-naringenin. This alternative cyclization would be accomplished by rotation of the coumaroyl moiety outward towards the solvent accessible active site entrance. However, Leu 38 and Lys 109 constrain the orientation of the coumaroyl moiety in the binding cleft. Architecturally, the CHI active site limits the substrate's available conformations to ensure stereospecific product formation.
Subtle variations in substrate preference reflected in the Km values for chalcone versus 6′-deoxychalcone exist between CHIs of different species. CHIs from legumes, such as alfalfa, prefer 6′-deoxychalcone as a substrate but the enzymes from non-legumes, like petunia, optimally use chalcone. The structure of the CHI•naringenin complex (SEQ ID NO:9), viewed with reference to the amino acid sequences of different CHIs (SEQ ID NOs:1-8), suggests that Thr 190 and Met 191 may partially modulate substrate preference. In the CHIs from non-legumes, a serine and an isoleucine replace Thr 190 and Met 191, respectively. These two differences may better accommodate the 6′-hydroxyl moiety of chalcone due to a modest increase in active site volume in the vicinity of the trihydroxyl ring.
Catalytic Mechanism
CHI catalyzes an intramolecular reaction utilizing a substrate-derived nucleophile and a carbon-carbon double bond as a Michael acceptor. Two reaction mechanisms have been proposed for (2S)-naringenin formation by CHI. One involves nucleophilic catalysis by an active site residue that forms a covalent intermediate that is released after a SN2 displacement by the 2′-O− of chalcone. The other mechanism invokes general acid-base catalysis employing an enol intermediate. The structure of CHI clearly supports the latter mechanism.
Examination of the CHI•naringenin complex structure (SEQ ID NO:9) reveals a hydrogen bond network at the bottom of the binding cleft centered about the water molecule that contacts the ketone of (2S)-naringenin (FIGS. 4A and 4B). Of the five amino acids contributing to this network, only Thr 48 and Tyr 106 are conserved in all CHIs (SEQ ID NOs:1-8). The position of the water molecule between (2S)-naringenin and Tyr 106 (SEQ ID NO:9) suggests a reaction mechanism in which the tyrosine activates the water, allowing it to serve as a general acid in the cyclization reaction (FIG. 4C). In the proposed reaction mechanism, the 2′-O− (pKa ̃7-8) forms in solution as suggested by studies on the spontaneous cyclization of chalcones. The negatively charged oxygen then attacks the carbon-carbon double bond utilizing a Michael addition with the water molecule at the backside of the active site acting as the general acid in the transient protonation of the intermediate enolate.
To test this reaction mechanism, Tyr 106 (SEQ ID NO:1) was substituted by phenylalanine and the properties of the mutant CHI (SEQ ID NO:13) compared to the wild-type enzyme. The kinetics for the cyclization of 6′-deoxychalcone by wild-type CHI (kcat=4384 min−1; Km=25.7 μM; kcat/Km=1.71×108M−1 min−1) versus those of the reaction catalyzed by the CHI Y106F mutant (kcat=69.0 min−1; Km=29.1 μM; kcat/Km=2.37×106 M−1 min−1) demonstrate that the tyrosine residue contributes to the stabilization of the transition state. The 100-fold reduction in reaction rate is consistent with the decrease in rate associated with the loss of a general acid. However, the observed reaction rate with the mutant remains greater than that of the uncatalyzed cyclization reaction. It is suggested that the structural complementarity of the binding cleft to the transition state of the reaction contributes additional levels of catalytic rate enhancement.
A major contribution to rate enhancement in enzymatic reactions results from bringing substrates or reactive centers in the same molecule together in space. As described above, the topology of the binding cleft limits the flexibility of chalcone and eliminates catalytically unproductive orientations by spatially defining an optimal geometry for (2S)-naringenin formation. This effectively channels the ground state conformation of the substrate into a catalytically productive conformation. Together with contributions from general acid-base catalysis, shape complementarity between the CHI binding pocket and chalcone accelerates the cyclization of chalcone 107-fold over the spontaneous reaction rate.







TABLE 4







Kinetic constants for wild-type and Y106F mutant CHI











kcat (min−1)
Km (μM)
kcat/Km (M−1 min−1)














CHI wild-type
4384 ± 517 
25.7 ± 5.4
1.71 × 108


(SEQ ID NO: 1)


CHI Y106F
69.0 ± 5.1 
29.1 ± 3.7
2.37 × 106


(SEQ ID NO: 2)









The three-dimensional structure of CHI, together with the structure of chalcone synthase, provides a useful template for engineering isomerases to develop, diversify and modify flavonoid biosynthetic pathways for crop and food sources, as well as providing novel flavanones for intermediates and leads in drug discovery. All figures were prepared with MOLSCRIPT or GRASP and rendered with POV-Ray.
While the foregoing has been presented with reference to particular embodiments of the invention, it will be appreciated by those skilled in the art that changes in these embodiments may be made without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims.
Table 5. PDB Accession No. 1EYQ (SEQ ID NO:9). The content of Table 5 is hereby incorporated by reference under 37 C.F.R. §1.52(e)(1)(iii) to file “1EYQ.txt” of CD-R disk “Tables”, created Jan. 2, 2007, having file size 342,918 bytes.
Table 6. PDB Accession No. 1FM7 (SEQ ID NO:9). The content of Table 6 is hereby incorporated by reference under 37 C.F.R. §1.52(e)(1)(iii) to file “1FM7.txt” of CD-R disk “Tables”, created Jan. 2, 2007, having file size 325,452 bytes.
Table 7. PDB Accession No. 1FM8 (SEQ ID NO:9). The content of Table 7 is hereby incorporated by reference under 37 C.F.R. §1.52(e)(1)(iii) to file “1FM8.txt” of CD-R disk “Tables”, created Jan. 2, 2007, having file size 315,366 bytes.
Table 8. PDB Accession No. 1JEP (SEQ ID NO:9). The content of Table 8 is hereby incorporated by reference under 37 C.F.R. §1.52(e)(1)(iii) to file “1JEP.txt” of CD-R disk “Tables”, created Jan. 2, 2007, having file size 340,294 bytes.

## Claims

1. A method for identifying a compound that binds to chalcone isomerase, said method comprising:
a) employing a three-dimensional (3-D) model of chalcone isomerase having the structural coordinates of Table 1, Table 5, Table 6, Table 7 or Table 8, to generate a 3-D model of a chalcone isomerase active site defined by the coordinates of residues 36, 37, 38, 47, 48, 49, 50, 97, 101, 106, 109, 110, 112, 113, 152, 190, 191, and 200 of SEQ ID NO:1,
b) designing or selecting a compound that potentially binds to said chalcone isomerase active site, and
c) contacting said compound with a chalcone isomerase in vitro and determining its ability to bind thereto, wherein said chalcone isomerase has at least 90% identity with respect to the sequence set forth in SEQ ID NO:1, and has chalcone isomerase enzymatic activity,

whereby compounds that bind chalcone isomerase are identified.

2. The method of claim 1, wherein said chalcone isomerase employed in step (c) is a mutant of a known chalcone isomerase, wherein said mutant has one or more conservative R-group modifications to the amino acids of a wild-type chalcone isomerase.

3. A method for identifying a compound that inhibits chalcone isomerase enzymatic activity, said method comprising:
a) employing a three-dimensional (3-D) model of chalcone isomerase having the structural coordinates of Table 1, Table 5, Table 6, Table 7 or Table 8, to generate a 3-D model of a chalcone isomerase active site defined by the coordinates of residues 36, 37, 38, 47, 48, 49, 50, 97, 101, 106, 109, 110, 112, 113, 150, 152, 190, 191, and 200 of SEQ ID NO:1,
b) designing or selecting a compound that potentially binds to said chalcone isomerase active site, and
c) contacting said compound with a chalcone isomerase in vitro and determining its ability to inhibit chalcone isomerase enzymatic activity, wherein said chalcone isomerase has at least 90% identity with respect to the sequence set forth in SEQ ID NO:1, and has chalcone isomerase enzymatic activity,

whereby compounds that inhibit the activity of chalcone isomerase are identified.

4. The method of claim 3, wherein said chalcone isomerase employed in step (c) is a mutant of a known chalcone isomerase, wherein said mutant has one or more conservative R-group modifications to the amino acids of a wild-type chalcone isomerase.