# Patent

## Title
Aromatic prenyltransferases, nucleic acids encoding same and uses therefor

## Abstract

In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture. The complexes with both aromatic substrates and prenyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and aromatic co-substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.

## Background
ACKNOWLEDGMENT
This invention was made with government support under Grant No. AI-051438 by the National Institutes of Health. The government has certain rights in the invention.


FIELD OF THE INVENTION
The present invention relates to aromatic prenyltransferases, nucleic acids encoding same, crystalline forms of aromatic prenyltransferases, and various uses therefor. In one embodiment, methods are provided for predicting the activity and/or substrate specificity of putative aromatic prenyltransferases. In another embodiment, methods of screening compounds to identify compounds which bind aromatic prenyltransferases and/or modulate the activity thereof, are provided. In yet another embodiment, methods of screening compounds to identify potential substrates of aromatic prenyltransferases are provided. In still another embodiment, methods are provided for prenylating aromatic structures, as well as controlling and/or modifying the degree of prenylation promoted by aromatic prenyltransferases. In a further embodiment, methods are provided for identifying proteins having the newly discovered beta/alpha barrel structure. In a still further embodiment, methods are provided for controlling and/or modifying the substrate specificity of aromatic prenyltransferases.
BACKGROUND OF THE INVENTION
Nature is a prolific producer of small molecules that have evolved to interact with diverse biological targets. From a human health perspective, natural products have dramatically altered our lives by providing many front-line drugs as well as chemical probes to unravel basic molecular pathways germane to health and disease. Although natural products continue to provide about half of all new chemical entities approved as drugs by the US Food and Drug Administration, drug discovery during the latter part of the 20th century shifted away from natural products towards synthetic libraries. This paradigm shift reflected the complexity of small, natural libraries against the simplicity of large, combinatorial synthetic libraries and was rationalized in order to keep pace with the enormous capacity of industrial high-throughput screening programs. New drugs from combinatorial chemical libraries, however, did not materialize during this time period, while natural products continued as an important source. Natural products, like drugs, cover a chemical space that is much more diverse than combinatorial compounds, thereby reflecting the rich chemical diversity of this resource.
Recent technological advances in natural product research involving isolation, characterization, synthesis, and biosynthesis have rekindled an interest in their investigation in academia and industry. With the advent of modern molecular biology, the field of biosynthesis has blossomed over the past decade with new approaches to generate biosynthetic libraries that further extend natural product structural diversity into new chemical space. In vivo approaches involving combinatorial biosynthesis, mutasynthesis, and precursor-directed biosynthesis and complementary in vitro approaches that combine chemical synthesis and enzymology (chemoenzymatic synthesis) have led to impressive libraries of novel molecules never encountered in nature. Natural product structural classes that have been biosynthetically manipulated in this fashion include the polyketides, nonribosomal peptides, terpenoids, and alkaloids. Most progress in this burgeoning field has resided with the actinomycetes (soil bacteria), which offer impressive arrays of natural products whose biosynthetic genes are typically clustered and are thus readily amenable to genetic manipulation. One notable exception that is absent from the biosynthetic diversification platform, however, is the hybrid isoprenoid class of natural products.
Natural products, such as the isoprenoid (terpenoid) family of diverse chemical scaffolds have held significant interest for the synthetic organic chemistry community because they are both challenging synthetic projects and possess varied biological activities and medicinal properties. Within the terpenoid family, the total synthesis of sesquiterpene natural products and related analogs continue to dominate the chemical literature. The demand for a reliable production platform for structurally complex terpenes has increased dramatically over the last 10 years and is of growing interest. Elegant synthetic schemes for terpenoids have been developed, but suffer from low yields and low regio- and enantio-selectivity. Although engineered E. coli has the potential to make mg/L levels of sesquiterpene hydrocarbons, the more biologically active terpenes are highly functionalized with hydroxyl, methyl, acetyl, halide, carbohydrate, and peroxide functional groups that require multi-step biosynthetic mechanisms often tethered to endo-membrane systems conducive for metabolic coupling. By integrating biosynthetic complexity with synthetic diversification, it may be possible for many of these hurdles to the technological development of terpenoids to be overcome.
Moreover, hybrid compounds containing terpene-derived residues comprise a large and diverse group of natural products that command an important role in human health (see Table 1). Historically this class of compounds has provided important drugs (e.g., the anticancer agent vincristine, the antimalarial quinine and the immunosuppressant mycophenolate mofetil) as well as challenging synthetic targets (e.g., strychnine and reserpine). In addition to natural products, many important coenzymes (ubiquinone and plastoquinone) and vitamins (tocopherols, phylloquinones, and menaquinones), which function in electron transport systems, contain isoprenoid residues.







TABLE 1







Representative hybrid isoprenoids, their


sources and biological significance










Natural

Isoprenoid



Product
Source
Hybrid
Biological Activity





mycophenolic
fungus
polyketide
immunosuppressant


acid


khellin
plant
polyketide
bronchial asthma


tetrahydro-
plant
polyketide
narcotic, antiemetic


cannabinol


rotenone
plant
isoflavonoid
insecticide


psoralen
plant
coumarin
skin pigment and irritant


novobiocin
bacterium
coumarin
antibiotic


lucidin
plant
quinine
mutagen


emetine
plant
tetrahydro-
alkaloid emetic (ipecac)




isoquinoline


ergometrine
fungus
ergot
alkaloidoxytocic


reserpine
plant
indole
alkaloidantihypertensive


vincristine
plant
indole alkaloid
anticancer


strychnine
plant
indole alkaloid
toxin


lyngbyatoxin
cyano-
indole alkaloid
inflammatory agent



bacterium


quinine
plant
quinoline alkaloid
antimalarial


camptothecin
plant
quinoline alkaloid
topoisomerase/inhibitor









Nature has assembled a myriad of scaffolds to which isoprenoids have been attached, and these include polyketides (the so-called meroterpenoids), flavonoids, coumarins, quinones, alkaloids, phenazines, and the like. Often the terpenoid unit is further elaborated by electrophilic cyclization and oxidative chemistry upon attachment to its building block, thereby leading to the great structural diversity observed within this group. While most of these natural products contain a single isoprenoid unit of varying chain length, others harbor multiple isoprene units such as in the tetraprenylated benzoylphloroglucinol derivatives sampsoniones A-I.
The vast majority of hybrid isoprenoids are derived from eukaryotes, particularly plants. For instance, over a thousand monoterpenoid indole alkaloids have been characterized, making this a major class of plant alkaloids. On the other hand, terpenoids, and in particular hybrid isoprenoids, appear to have a limited distribution in prokaryotes. While actinomycetes are metabolically very rich bacteria and produce many important biosynthetic classes of natural products that include polyketides, nonribosomal peptides, aminoglycosides, and the like, the terpenoids are notably scarce. As a consequence, while other natural product structural classes have been biosynthetically exploited in the drug discovery arena, the hybrid isoprenoids are noticeably absent due to our limited understanding of their biosynthesis at the biochemical and genetic levels.
The majority of the basic understanding of how hybrid isoprenoids are biosynthesized in plants, fungi and bacteria is based on feeding experiments with labeled precursors. Enzymes and their encoding genes associated with interfacing isoprenyl diphosphates with their small molecule building blocks are very few and are mostly associated with plant natural products such as shikonin and with coenzymes and vitamins such as the ubiquinones, plastoquinones, menaquinones, and tocopherols. Very recently, two prokaryotic prenyltransferases (PTases) involved in the biosynthesis of the streptomycete antibiotics clorobiocin and novobiocin and the cyanobacterial toxin lyngbyatoxin were discovered. These soluble, monomeric PTases contrast with the membrane-associated PTases previously identified from eukaryotes.
Actinomycetes produce a limited set of pure and hybrid terpenoids. The antibiotic novobiocin was the first streptomycete natural product discovered with a terpenoid side chain; this group has since grown to include other members bearing naphthoquinones (naphterpin, furaquinocin, napyradiomycins), phenazines (lavanducyanin, aestivophoenin), shikimate-derived quinones, and other aromatic substrates (see FIG. 1B). Feeding experiments delineated a number of biosynthetic pathways, including those to novobiocin, naphterpin, and furaquinocin, and revealed that actinomycetes utilize both the mevalonate and nonmevalonate (methyl-D-erythritol 4-phosphate (MEP)) pathways to synthesize their isoprene building blocks.
The development of novel methodologies related to natural products chemistry and biosynthesis is of growing interest. Prenylated aromatic natural products appear to be a very promising class of therapeutically compounds. The prenylation of aromatic compounds often leads to significant alteration in the bioactivity profile of a compound, by both the creation of a novel C—C bond and also the introduction of one or more double bonds in the framework of the final product. Such compounds can affect a wide variety of biological systems in mammals and include roles as antioxidants, anti-inflammatories, anti-virals, anti-proliferatives, and anti-cancers.
Prenyltransferases (PTases) are ubiquituous enzymes that catalyze the alkylation of electron rich prenyl acceptors by the alkyl moieties of allylic isoprene diphosphates. Prenyltransferases utilize isoprenoid diphosphates as substrates, and catalyze the addition of the acyclic prenyl moiety to isopentenyl diphosphate (IPP), higher order prenyl diphosphates, aromatic rich molecules and proteins. Until now, only a few “aromatic” prenyltransferases have been isolated, each of which has been shown to interact with only a limited range of substrate(s) and/or prenyl donors. Such prenyltransferases have otherwise only been nominally characterized; and none of such prenyltransferases have been characterized at the structural level.
Accordingly, there is a need in the art for the identification of novel enzymes capable of promoting the prenylation of aromatic compounds, as well as compounds which can modulate the prenylation of aromatic compounds. These and other needs are addressed by the present invention, as described in greater detail in the specification and claims which follow.
SUMMARY OF THE INVENTION
In accordance with the present invention, a novel aromatic prenyltransferase, Orf2 from Streptomyces sp. strain CL190, involved in naphterpin biosynthesis (Shin-ya, et al., in J. Antibiot. (Tokyo) 43, 444-447 (1990)) has been identified and the structure thereof elucidated. This prenyltransferase catalyzes the formation of a C—C bond between a prenyl group and a compound containing an aromatic nucleus, and also displays C—O bond formation activity. Numerous crystallographic structures of the prenyltransferase have been solved and refined, e.g., (1) prenyltransferase complexed with a buffer molecule (TAPS), (2) prenyltransferase as a binary complex with geranyl diphosphate (GPP) and Mg2+, and prenyltransferase as ternary complexes with a non-hydrolyzable substrate analogue, geranyl S-thiolodiphosphate (GSPP) and either (3) 1,6-dihydroxynaphthalene (1,6-DHN), or (4) flaviolin (i.e., 2,5,7-trihydroxy-1,4-naphthoquinone, which is the oxidized product of 1,3,6,8-tetrahydroxynaphthalene (THN)). These structures have been solved and refined to 1.5 Å, 2.25 Å, 1.95 Å and 2.02 Å, respectively. This first structure of an aromatic prenyltransferase displays an unexpected and non-canonical (β/α)-barrel architecture.
The complexes with both aromatic substrates and geranyl containing substrates and analogs delineate the active site and are consistent with a proposed electrophilic mechanism of prenyl group transfer. These structures also provide a mechanistic basis for understanding prenyl chain length determination and substrate recognition in this structurally unique family of aromatic prenyltransferases. This structural information is useful for predicting the aromatic prenyltransferase activity of proteins.
Specifically, the present disclosure describes the identification of two novel aromatic prenyltransferases with promiscuous activity: Orf2 from Streptomyces CL.190 and HypSc from Streptomyces coelicolor. The present disclosure also describes a high resolution structure of a new type of β/α-barrel which provides a useful structural template for understanding the mechanistic features accompanying Orf2's promiscuous activity with respect to a number of aromatic prenyl acceptors and its means of regulating prenyl chain length specificity through a well ordered prenyl chain binding surface. The β/α-barrel catalyzes the prenylation of aromatic compounds, accepts a wide range of aromatic substrates and uses hydrophobic interactions to bind the hydrocarbon moiety of an allylic diphosphate substrate (GPP or FPP).
It is demonstrated herein that this “biosynthetic barrel” can be used as starting point for engineering the prenylation of natural products of both microbial and plant origin. The structural details involved in substrate specificity in this newly characterized small molecule prenyltransferase enables the biosynthetic diversification of numerous aromatic compounds foind in nature, and of synthetic origin by providing a structurally guided process of enzyme design and evolution, leading to the production and metabolic engineering of novel prenylated natural products through in vivo transgenic approaches, or ultimately, for in vitro combinatorial chemistry.



BRIEF DESCRIPTION OF THE FIGURES
FIG. 1A presents the structures of hybrid terpenoid-polyketide compounds produced by Actinomycetes. The synthesis of naphterpin involves the prenylation of THN, flaviolin or a derived metabolite using a GPP co-substrate. THN is produced from malonyl-CoA by the action of THN synthase encoded by orf3. THN is readily oxidized to give a hydroquinone derivative, 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin). The THN skeleton is further modified, prenylated and incorporated into hybrid terpenoid-polyketide compounds such as naphterpin, furaquinocin A, napyradiomycin A and marinone.
FIG. 1B provides structures of representative hybrid isoprenoids from actinomycetes. Isoprenoid units are appended to naphthoquinone (see naphterpin, marinone, neomarinone, and Q525.518), phenol (see novobiocin), phenazine (see lavanducyanin and aestivophoenin B), and nitropyrrole (see Q509.364) residues via C-, N-, and O-linkages, as appropriate.
FIG. 1C presents a structure based multiple sequence alignment. The orf2 gene product of SEQ ID NO:2 from Streptomyces sp. strain CL190, Orf2, is a 33kDa soluble, monomeric protein comprising 307 residues. PSI-BLAST searches revealed strong homologies between Orf2 and three other bacterial proteins: a protein from Streptomyces coelicolor A3(2)(HypSc, accession number AL939130) (SEQ II) NO: 5) and the previously described 4-hydroxyphenylpyruvate: dimthyallyl transferase genes, cloQ (accession number AF329398) (SEQ ID NO: 7) and novQ (accession, number AF170880) (SEQ ID NO: 6), from Streptomyces roseochromogenes and Streptomyces spheroids NCIMB 11891, respectively. Residues (one-letter amino acid code) are numbered according to Orf2's sequence. Dashes represent insertions and deletions. This alignment has been linked with the known Orf2 secondary structure and rendered with ESPript (accessible via the internet on the world wide web at the URL “prodes.toulouse.inra.fr/ESPript”). The coding is as follows: grey on grey for residues located in the active site, white on black for residues strictly conserved, and white on grey overlay for residues both strictly conserved and located in the active site. Residues bounded by grey frames represent similar residues in the aligned sequences.
FIG. 2A illustrates the Mg2+ dependent prenylation of 1,6-DHN. The reaction buffer consisted of 50 mM HEPES (pH 7.5), 5 mM 1,6-DHN, and 5 mM GPP in a final volume of 20 μl. The reaction was initiated by adding 20 μg of Orf2 to the assay mixture. After incubation at 25° C. for 4 hours, the mixture was dried, and spotted on a silica gel TLC plate. The TLC plate was developed with a chloroform/methanol (20:1) solvent mixture. 1,6-DHN and reaction products were detected at 254 nm. The chemical analyses of the two HPLC purified products were accomplished by both MS and 1H NMR analyses. In lane 1 (control), Orf2 was boiled prior to adding. The reaction mixture in lane 2 contained no MgCl2, while 5 mM MgCl2 was added in lane 3.
FIG. 2B illustrates the promiscuous activity of Orf2. Several assays were conducted, employing a variety of potential substrates, i.e., 1,3-DHN (1), 1,6-DHN (2), 2,7-DHN (3), 4-HPP (4) and several isoflavonoids and polyketide derivatives, including daidzein (7,4′-dihydroxyisoflavonone, 5), formononetin (7-hydroxy, 4′-methoxyisoflavonone, 6), fisetin (3,3′,4′,7-tetrahydroxyflavone, 7), genistein (5,7,4′-trihydroxyisoflavone, 8), naringenin (5,7,4′-trihydroxyflavonone, 9), flaviolin (10), olivetol (11), olivetolic acid (12), and resveratrol (3,4′,5-trihydroxystilbene, 13). The chemical structures of four reaction products (i.e., 1,6-dihydroxy 2-geranyl naphthalene (14), 1,6-dihydroxy 5-geranyl naphthalene (15), 6-geranyl naringenin (16) and 7-O-geranyl naringenin (17)) were determined by both MS and 1H NMR analyses. The reaction buffer consisted of 50 mM HEPES (pH 7.5), 5 mM MgCl2, and 0.1 mM GPP, 0.009 mM [14C]GPP, and 0.1 mM of each substrate, in a final volume of 20 μl. The reaction was initiated by adding 30 μg of Orf2 to the assay mixture. After incubation at 25° C. for 6 hours, the mixture was dried, and spotted on a silica gel TLC plate. The TLC plate was developed with a chloroform/methanol (15:1) solvent mixture. Reaction products were detected with a [14C] imaging plate (Fuji Photo Film).
FIG. 3 collectively presents a comparison of the different types of protein barrel topologies. Two-dimensional topology diagrams and three dimensional views of protein barrels are displayed from top to bottom. Each secondary structure element (helices represented as circles (or spiral ribbons) and β-strands as triangles (or flat ribbons)) maintains directionality (N to C) which is either “up” (out of the plane of the diagram) or ‘down’ (into the plane of the diagram). The direction of elements can be deduced from the connecting lines, and also from the orientation of the strands.
FIG. 3A illustrates an α/β-barrel (e.g., an human aldo-keto reductase complexed with NADP+ and glucose 6-phosphate (pdb entry 2ACQ)).
FIG. 3B illustrates a β/α-barrel (e.g., Streptomyces sp. strain CL190 Orf2 aromatic prenyltransferase of SEQ ID NO:2 complexed with GSPP, DHN2 and Mg+2).
FIG. 3C illustrates an α+β-barrel (e.g., a dimeric ferrodoxin-like α+β sandwich fold of the ActVA-Orf6 monooxygenase (pdb entry 1LQ9) from S. coelicolor A3).
FIG. 3D illustrates a β-barrel (e.g., human fatty acid binding protein, M-FABP, complexed with one molecule of stearic acid (pdb entry 1HMT).
FIG. 3E illustrates an α-α barrel (e.g., the β-subunit of the Rattus norvegicus protein farnesyltransferase complexed with farnesylated Ras4B peptide product and farnesyl diphosphate substrate bound simultaneously (pdb entry 1KZO)).
FIG. 4 collectively presents close-up views of the active site of Orf2 of SEQ ID NO:2 in different complexes.
FIG. 4A illustrates the following complexes: the TAPS molecule, bound GPP, and GSPP with either 1,6-DHN or flaviolin.
FIG. 4B illustrates the structure of the divalent metal binding site. A representative 2fo-fc electron density map (rendered from a normalized map at 1.0σ level) displays octahedral coordination of the Mg2+ ion, where two oxygen atoms, one from Asp 62, and one from the diphosphate moiety of the GSPP molecule contribute together with four water molecules to the octahedral coordination geometry.
FIG. 4C provides a schematic representation of the active site of Orf2 of SEQ ID NO:2 . The side chain involved in Mg2+, GSPP and 1,6-DHN binding is depicted with hydrogen and coordination bonds as grey dashed lines. Black dashed lines represent indirect hydrogen bonds via a water molecule. This close-up view, shown in an identical orientation to that in FIG. 4A, is rotated by 180 degrees along the vertical axis compared to that depicted in FIG. 4B. The half circles depict van der Waals contacts with the two substrates. Tentative depth queuing coding is as follows: grey for residues in the back of the GSPP-1 ,6-DHN plane, black in the same plane, and thick black for residues in the front.
FIG. 5 presents a schematic and structural representation of the proposed mechanism for aromatic prenylation in the active site of Or2 of SEQ ID NO:2 . This panel depicts the binding of the aromatic substrate next to the GPP molecule, formation of a geranyl carbocation (noted G+), rotation of the prenyl chain into a productive conformation, electrophilic attack of the carbocation on the aromatic ring of 1,6-DHN, formation of a a-complex, and final proton removal by a water molecule.
FIG. 6 presents active site models for Orf2 homologs. Modeling of CloQ/NovQ and HypSc were performed using Orf2 as a structural template. Side chains presenting potentially significant variation between the different active sites are displayed and labeled. Conserved residues in the different models include Asp 110, Lys 119, Asn 173, Tyr 175, Tyr 216, and Arg 228, of which only Asp 110 and Arg 228 are displayed for clarity.
FIG. 7 relates to the functional evaluation of HypSc. Thus, HypSc prenyltransferase activity was assayed as described above with respect to FIG. 2A, using 1,6-DHN as a prenyl acceptor (in each of lanes 1-4), with no prenyl acceptor in lane 1, DMAPP in lanes 2 and 3, and GPP in lane 4. No Mg2+ was used in lane 2. The samples were incubated overnight at room temperature.
FIG. 8 is a block diagram of a computer system contemplated for use in the practice of the present invention.



DETAILED DESCRIPTION OF THE INVENTION
Naphterpin is a bioactive natural product (hemiterpenoidal anti-oxidant agent) produced by Streptomyces sp. strain CL190 via both the mevalonate (MVA) isoprenoid biosynthetic pathway as well as a polyketide biosynthetic pathway (see, for example, Shin-ya, et al., in Tetrahedron Lett. 31, 6025-6026 (1990); Shin-ya, et al., in J. Antibiot. (Tokyo) 43, 444-447 (1990); and Seto, et al, in Tetrahedron Letters 37(44):7979 (1996), see also FIG. 1A). The compound is composed of a tetrahydroxynaphthalene (THN) derivative and a geranyl moiety. THN is known in the art to be biosynthesized from 5 molecules of malonyl coenzyme A (CoA) by the action of type III polyketide synthase (THN synthase) cloned from Streptomyces griseus (see Funa, et al., in Nature 400, 897-899 (1999)) and Streptomyces coelicolor (see Izumikawa et al., in J. Ind. Microbiol. Biotechnol. 30:510-515 (2003)). Compounds with naphthoquinone rings, including naphterpin, furaquinocin, napyradiomycin and marinone, are biosynthesized via the symmetric polyketide intermediate 1,3,6,8-tetrahydroxynaphthalene (THN; see Shin-ya, et al., in J. Antibiot. (Tokyo) 43, 444-447 (1990)) (FIG. 1A). In Streptomyces griseus and Streptomyces coelicolor A3(2), THN is the product of a chalcone synthase-like type III polyketide synthase (PKS), known as THN synthase (THNS) (Austin and Noel, Nat Prod Rep 20(1):79-110 (2003). THN readily (or enzymaticaly) oxidizes forming a hydroquinone derivative, 2,5,7-trihydroxy-1,4-naphthoquinone (flaviolin), part of which subsequently undergoes polymerization to form a variety of colored polymeric compounds (Funa et al., Nature 400(6747):897-9 (1999)).
In addition to its role in pigment production, the THN skeleton is further modified and incorporated into naphterpin in Streptomyces sp. strain CL190 (Shin-ya et al., J. Antibiot (Tokyo) 45(1):124-5 (1992)).
In actinomycetes, three mevalonate gene clusters have been cloned to date, i.e., from CL190, Kitasatospora griseola (terpentecin producer) (see Hamano, et al., in Biosci. Biotechnol. Biochem. 65:1627-1635 (2001)), and Actinoplanes sp. strain A40644 (BE-40644 producer) (see Kawasaki, et al., in J. Antibiot. 56:957-966 (2003)). All of these clusters encode mevalonate kinase, diphosphomevalonate decarboxylase, phosphomevalonate kinase, isopentenyl diphosphate isomerase, 3-hydroxy-3-methylglutaryl (HMG)-CoA reductase, and HMG-CoA synthase. The order of each of the genes is also the same and respective homologous genes have 50 to 80% amino acid identity with each other.
In contrast to the high conservation of the mevalonate pathway gene clusters, a diversity of genes is distributed in their flanking regions. For example, the geranylgeranyl diphosphate synthase, a key enzyme of the terpentecin biosynthesis, is encoded in the region just upstream of the mevalonate kinase gene, with the terpentecin biosynthetic gene cluster located further upstream. In addition, farnesyl diphosphate synthase, a key enzyme of the BE40644 biosynthesis, is located just upstream of the mevalonate kinase gene, with the BE-40644 biosynthetic gene cluster located in the region downstream of the mevalonate pathway gene cluster.
These facts, taken together, gave rise to the hypothesis that the mevalonate pathway genes cluster, that terpenoid biosynthetic genes are usually clustered in a terpenoid-producing actinomycetes, and that the mevalonate pathway gene cluster could be a good marker to clone the terpenoid biosynthetic genes from the terpenoid-producing actinomycetes. Based on this hypothesis, in order to clone a naphterpin biosynthetic genes cluster, the flanking regions of the mevalonate pathway genes cluster which was cloned from CL190 were sequenced.
To understand the biosynthetic pathway of this mixed terpene/polyketide derived natural product, the gene cluster responsible for naphterpin production was identified based upon proximity to genes encoding the MVA pathway biosynthetic enzymes. An upstream region of the gene cluster containing the MVA pathway genes revealed three new open reading frames or orfs designated orf1, orf2, and orf3. The comparative analysis of these orfs with genes encoding functionally characterized proteins is summarized in Table 2. PSI-BLAST searches revealed homologies between Orf2 and three other bacterial proteins: a protein from Streptomyces coelicolor A3(2) (HypSc, accession number AL939130) and the previously described 4-hydroxyphenylpyruvate:dimethylallyl transferase genes, cloQ (accession number AF329398) and novQ (accession number AF170880), from Streptomyces roseochromogenes and Streptomyces spheroides NCIMB 11891, respectively (FIG. 1B).
To further understand the. function of the genes referred to above, a mutant Streptomyces sp. (strain CL190) was prepared by disrupting the Orf2 gene of SEQ ID NO:1. This mutant exhibited no naphterpin production. The high degree of homology between Orf2 and the functionally characterized prenyltransferases CloQ/NovQ (Pojer et al., Proc Natl Acad Sci USA 100:2316-2321 (2003)) (FIG. 1B), and the fact that Orf3 encodes a type III polyketide synthase with amino acid similarity to THNS, establishes that off2 encodes a prenyltransferase involved in geranyl group transfer to THN or a THN derivative produced through the action of orf3 (and possibly other tailoring enzymes).
When expressed in E. coli, Orf2 of SEQ ID NO:2 is as a 33kDa soluble, monomeric protein having 307 residues. To assess enzyme activity, the purified recombinant Orf2 protein was incubated with one of the following prenyl (geranyl) donors, i.e., dimethylallyl diphosphate (DMAPP, C05), geranyl diphosphate (GPP, C 10) or farnesyl diphosphate (FPP, C 15) along with several possible substrates (i.e., prenyl acceptors) possessing one or more aromatic groups. A variety of THN analogues (e.g.,1 ,3-dihydroxynaphthalene (1 ,3-DHN), 1 ,6-DHN, 2,7-DHN, and flaviolin) are observed to function as substrates for Orf2 of SEQ ID NO:2, i.e., are converted by Orf2 into prenylated derivatives thereof (see FIGS. 2A and 2B). The 4-hydroxyphenylpyruvate (4-HPP) substrate of CloQ/NovQ (Pojer et al, supra) was also converted by Orf2 of SEQ ID NO:2 into a prenylated derivative thereof. In contrast, the related molecules, phenylalanine or tyrosine, did not serve as substrates (see FIG. 2B). No activity was observed with DMAPP, the highest relative activity was observed with GPP, and weak activity was observed with FPP. In summary, Orf2 of SEQ ID NO:2 recognizes a variety of substrates.
Moreover, significant Mg2+ dependent, in vitro activity is observed with the dihydroxy containing THN analogs (FIGS. 2A and 2B). Thus, two prenylated products, 1,6-DHN-P1 and 1,6-DHN-P2, were readily detected by thin layer chromatography when Orf2 was incubated with 1,6-DHN and GPP (TLC, FIGS. 2A and 2B). Large scale incubations with GPP and 1,6-DHN produced a sufficient amount of both products (in an approximate ratio of 10:1) to permit their structure elucidation by both MS and 1H NMR analyses: these compounds, trans-5-geranyl 1,6-DHN and trans-2-geranyl 1,6-DHN, are believed to be novel natural products (see FIG. 2A).
Orf2's potential to serve as a template for the diversification of novel aromatic natural products was demonstrated by assaying the ability of Orf2 to interact with various flavonoids, isoflavonoids and related compounds (e.g., resveratrol; see FIG. 2B). While Orf2 showed prenyltransferase activity in the presence of daidzein (7,4′-dihydroxyisoflavanone), formononetin (7-hydroxy, 4′-methoxyisoflavanone), genistein (5,7,4′-trihydroxyisoflavone), and resveratrol (3,4′,5-trihydroxystilbene), little or no activity was observed in the same test conditions with fisetin (3,3′,4′,7-tetrahydroxyflavone). In the presence of naringenin (5,7,4′-trihydroxyflavanone) and GPP, two reaction products, 6-geranyl naringenin, and 7-O-geranyl naringenin, were identified (by both MS and 1H NMR analyses; see FIG. 2B). 6-geranyl naringenin (also known as bonannione A; see Bruno, Heterocycles 23(5):1147-1153 (1985)), is a prenylated flavanone displaying significant antibacterial activity (Schutz, Phytochemistry 40:1273-1277 (1995)). 7-O-geranyl naringenin, which harbors a prenyl unit in the form of an ether moiety, which is only occasionally found in isoflavones, is a novel prenylated flavonoid.
Only a trace component in hops, 6-geranyl naringenin is formed by the isomerization (cyclization) of the more abundant hop flavonoid, 2′,4′,6′,4-tetrahydroxy-3′-geranylchalcone. Interestingly, the antifungal activity of various yellow lupin constituents has been reported, using Cladosporium herbarum as the test fungus. It was found that for isoflavones, the 6-prenyl and 3′-prenyl compounds were more fungitoxic than the 8-prenyl analogues and that transformation of the prenyl group to a cyclized derivative greatly reduced or eliminated the fungitoxic effects.
Orf2 was also active in the presence of both olivetol and olivetolic acid (see FIG. 2B). These compounds are intermediates in the biosynthesis of the therapeutic plant derived polyketide-terpene natural product Δ9-tetrahydrocannabinol (Δ9-THC). Δ9-THC is the primary psychoactive component found in Cannabis sativa. A synthetic analogue thereof, i.e., dronabinol, is currently used to alleviate nausea/vomiting and to stimulate appetite in order to counter weight loss in cancer and AIDS patients. The primary psychoactive component in cannabis, Δ9-THC affects the brain mainly by activating two specific cannabinoid receptors (CB1 and CB2). These receptors also bind to ‘endogenous’ cannabinoids, which are produced naturally by the human body. Recent studies of the cannabinoid signaling system shows its involvement in an ever-increasing number of pathological conditions. As the geranyl prenyltransferase activity involved in Δ9-THC biosynthesis in Cannabis sativa has until now only been detected in cell extracts, it was decided to test Orf2's activity in the presence of both olivetol and olivetolic acid, two supposed intermediates of Δ9-THC biosynthesis: Orf2's reaction products were detected on TLC with both Δ9-THC precursors, differing from the C. sativa endogenous enzyme for which activity was only observed in the presence of olivetolate molecule. These results are very promising for the opening of new therapeutic avenues based on the ability to modulate the endocannabinoid system.
Thus, in accordance with the present invention, there are provided aromatic prenyltransferases having a beta/alpha barrel structure.
As used herein, the phrase “beta/alpha barrel structure” refers to a closed β-sheet comprising antiparallel β-strands arranged around a central β-barrel core, itself surrounded by a ring of α-helices forming the outer, solvent exposed surface of the beta/alpha barrel, as described in greater detail herein. Thus, aromatic prenyltransferases are seen to have the unique beta/alpha barrel secondary structure.
This protein is the first identified and structurally characterized enzyme involved in a mixed polyketide-isoprenoid biosynthetic pathway, namely naphterpin biosynthesis. While this protein family has been identified and characterized from Streptomyces bacteria, numerous prenylated aromatic natural products are found in plants. For example, the therapeutically important natural product, tetrahydrocannabinol (THC) is a mixed polyketide-isoprenoid. Biosynthetic logic would dictate that plants are likely to contain enzymes similar in structure and function to Orf2, but such enzymes have thus far not been identified. Given this likelihood, Orf2/CloQ/NovQ/HypSc are believed to be the first identified members of a widespread and catalytically interesting family of enzymes.
Exemplary aromatic prenyltransferases according to the present invention have the amino acid sequence set forth in SEQ ID NO:2, or conservative variations thereof, provided that the variant polypeptide retains prenyltransferase activity. As used herein, “conservative variations” refer to 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 amino acid.
Modifications and substitutions contemplated herein 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 (e.g., by deletion, replacement, or addition). Examples of such other modifications include incorporation of rare amino acids, dextra-amino acids, glycosylation sites, cytosine for specific disulfide bridge formation. The modified peptides can be chemically synthesized, or the isolated gene can be site-directed mutagenized, or a synthetic gene can be synthesized and expressed in bacteria, yeast, baculovirus, tissue culture, and the like.
Aromatic prenyltransferases having sequence substantially identical to the amino acid sequence set forth in SEQ ID NO:2 are also contemplated herein. By “substantially identical” is meant a polypeptide or nucleic acid exhibiting at least 50%, preferably 60%, more preferably 70%, more preferably 80%, more preferably 85%, more preferably 90%, and most preferably 95% homology to a reference amino acid or nucleic acid sequence, provided that the “substantially identical” polypeptide retains prenyltransferase activity.
Alternatively, aromatic prenyltransferases according to the present invention have at least 80% sequence identity with the amino acid sequence set forth in SEQ ID NO:2. Sequence homology and identity are 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). The term “identity” in the context of two or more nucleic acids or polypeptide sequences, refers to two or more sequences or subsequences that are the same or have a specified percentage of amino acid 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. The term “homology” in the context of two or more nucleic acids or polypeptide sequences, refers to two or more sequences or subsequences that are homologous or have a specified percentage of amino acid residues or nucleotides that are homologous 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. Programs as mentioned above allow for substitution of an amino acid with a similar amino acid by determining a degree of homology between the sequences being compared.
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 (typically having from about 20 up to about 600 contiguous residues) in which a sequence may be compared to a reference sequence of the same number of contiguous residues 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. Natl. 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, accessible on the world wide web (www) at the URL “weber.u.Washington.edu/ ̃roach/human_genome_progress 2.html”) (Gibbs, 1995). Several databases containing genomic information annotated with some functional information are maintained by different organization, and are accessible via the internet on the world wide web (www), for example, at the URL “tigr.org/tdb”; “genetics.wisc.edu”; “genome-www.stanford.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., Nucl. 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 on the world wide web (www) at the URL “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., Matrices for Detecting Distance Relationships: Atlas of Protein Sequence and Structure, Washington: National Biomedical Research Foundation (1978)). BLAST programs are accessible through the U.S. National Library of Medicine, e.g., accessible on the world wide web (www) 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.
In accordance with another aspect of the present invention, there are provided nucleic acids encoding any of the above-described prenyltransferases, including all variations embraced by the degeneracy of the genetic code. Exemplary nucleic acids according to the present invention include nucleic acids which specifically hybridize to the nucleotide sequence set forth in SEQ ID NO:1 (or the complement thereof) under stringent hybridization conditions, wherein said nucleic acid encodes an aromatic prenyltransferase.
Hybridization methods are well known to those skilled in the art of molecular biology. “Specifically hybridizable” and “specifically complementary” are terms that indicate a sufficient degree of complementarity such that stable and specific binding occurs between a first nucleic acid and a DNA or RNA target. The first nucleic acid need not be 100% complementary to its target sequence to be specifically hybridizable. A first nucleic acid is specifically hybridizable when there is a sufficient degree of complementarity to avoid non-specific binding of the first nucleic acid to non-target sequences under conditions where specific binding is desired. Such binding is referred to as specific hybridization.
An alternative indication that two nucleic acid molecules are closely related is that the two molecules hybridize to each other. In certain embodiments, orf2 nucleic acid variants hybridize to a disclosed orf2 nucleic acid sequence (or fragments thereof), for example, under low stringency, moderate stringency, or high stringency conditions. Hybridization conditions resulting in particular degrees of stringency will vary depending upon the nature of the hybridization method of choice and the composition and length of the hybridizing nucleic acid sequences. Generally, the temperature of hybridization and the ionic strength (for example, the Na+ concentration) of the hybridization buffer will determine the stringency of hybridization, although wash times also influence stringency. Calculations regarding hybridization conditions required for attaining particular degrees of stringency are discussed by Sambrook et al. (ed.), Molecular Cloning: A Laboratory Manual, 2nd ed., vol. 1-3, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989, chapters 9 and 11.
The following exemplary sets of hybridization conditions are not meant to be limiting. High stringency conditions include hybridization in 5×SSC at 65° C. for 16 hours, two washes in 2×SSC at room temperature (RT) for 15 minutes each and two washes in 0.5×SSC at 65° C. for 20 minutes each. Moderate stringency conditions include hybridization in 5×-6×SSC at 65° C.-70° C. for 16-20 hours, two washes in 2×SSC at RT for 5-20 minutes each and two washes in 1×SSC at 55° C.-70° C. for 30 minutes each. Low stringency conditions include hybridization in 6×SSC at RT to 55° C. for 16-20 hours and two washes in 2×-3×SSC at RT to 55° C. for 20-30 minutes each.
Alternatively, nucleic acids according to the present invention include nucleic acids having at least 80% sequence identity with the nucleotide sequence set forth in SEQ ID NO:1, wherein said nucleic acid encodes an aromatic prenyltransferase.
To investigate the structural features accompanying prenyl chain length determination, aromatic substrate selectivity and the mechanism of prenyl group transfer, X-ray crystal structures of four Orf2 substrate/substrate analogue complexes were determined, namely Orf2 complexed with a TAPS buffer molecule, a binary Orf2 complex containing GPP and Mg2+, a ternary Orf2 complex with a non-hydrolyzable GPP analogue (GSPP), Mg2+ and 1,6-DHN, and a ternary Orf2 complex with GSPP, Mg2+ and flaviolin (results are summarized in Table 3).
The three dimensional structure of Orf2 consists of a single domain that forms a novel barrel type of structure (FIG. 3). This new barrel, here termed a β/α-barrel, is a closed β-sheet comprising sufficient antiparallel β-strands to form a central β-barrel core (typically in the range of about 6 up to 12 β-strands), with the central β-barrel core surrounded by a ring of α-helices forming the outer, solvent exposed surface of the barrel (FIG. 3B). In the specific example when a β/α-barrel structure comprises 10 β-strands, the secondary connectivity nearly conforms to a (ααββ)5 classification, but is more specifically described using the (ααββ)4-(αββ)-α nomenclature, where helices 6 and 8, both involved in inter-protein contacts in the crystal lattice, display a helical “kink”.
The most hydrophobic section of the β/α-barrel is the region residing between the outer surface of the cylindrical β-barrel and the belt of surrounding α-helices. Additionally, a number of hydrophobic residues located inside the barrel accommodate the prenyl tail of the GPP and GSPP molecules, while the diphosphate or the thio-diphosphate head groups of substrate and substrate analogs, respectively, point toward the “upper”, more polar end of the barrel where a Mg2+ ion is coordinated. Typically, the bottom of the barrel is capped by a short C-terminal helix. In the specific example when a β/α-barrel structure comprises 10 β-strands, the C-terminal helix would be α11.
Structurally related proteins belonging to either the TIM barrel or the β-barrel structural families, both of which display barrel folds with connectivity patterns that are distinctively different from the β/α-barrel are illustrated herein (see FIG. 3). TIM barrel proteins (ie. the aldo-keto reductase family represented by pdb entry code 2ACQ) consist of a repeated β-strand-loop-α-helix-loop motif, most often containing eight repeats, with the parallel β-strands forming the interior of an open barrel, and the helices forming the outer belt of the complete protein (Gerlt and Raushel, Curr Opin Chem Biol 7(2):252-64 (2003)) (FIG. 3A).
β-barrel proteins including human fatty acid-binding proteins (FABP, pdb entry code 1HMT), consist of ten anti-parallel β-strands arranged as an elliptical barrel capped at the bottom by two short α-helices (Sacchettini et al., J Mol Biol 208(2):327-39 (1989); Xu et al., J Mol Biol 268(11):7874-84 (1993) (FIG. 3D).
Another class of protein displaying an elliptical β-barrel surrounded by helices is the dimeric ferredoxin-like α+β sandwich fold: the ActVA-Orf6 monooxygenase (pdb entry code 1LQ9) from S. coelicolor A3 belongs to the latter class, and is a small enzyme that oxidizes a relatively large three ringed aromatic substrate at two active sites located between β-sheets and α-helices (FIG. 3C).
α/β-barrels have been defined as large structures (at least 200 amino acids), predominantly composed of alternating α-helices and β-strands, with parallel β-strands forming a “hub” surrounded by a “tire” of α-helices (see Branden and Tooze, Introduction to protein structure. Second edn. (1999), New York: Garland), while α+β class encompasses proteins with mainly antiparallel β-sheets but with segregated α-helical and β-sheet regions. In this regard, it is proposed that the α/β class definition, including protein domains exclusively composed of parallel β-strands, connected by α-helices, should be enlarged to include Orf2's novel architecture, the β/α-barrel. The β/α-barrel would introduce a novel β/α-barrel category comprising antiparallel β-strands connected and surrounded by α-helices as a subcategory of the α/β-class, but distinct from the α/β-barrel subcategory.
Interestingly, a last type of barrel, an α-α barrel domain, can be found in the β-subunit of the heterodimeric human protein farnesyltransferase, which catalyzes the carboxyl-terminal prenylation of Ras and several other signaling proteins (Park et al., Science 275(5307):1800-4 (1997) (FIG. 3D). This domain displays a very different overall fold but presents a similar aromatic rich substrate binding pocket and active site topology as described herein for Orf2 (Park et al., supra; Long et al., Nature 419(6907):645-50 (2002)).
It seems, indeed, that isoprenyl diphosphate synthases, protein prenyltransferases, and prenyltransferases (PTases), all involved in the binding of prenyl compounds, use a similar strategy regarding the active site environment. In most cases, prenyl chain bonding occurs within a large hydrophobic tunnel with highly conserved residues. Structures of the trans-type farnesyl diphosphate synthase display two identical subunits associated as a homodimer, forming a four layer helix-bundle; eight of these helices are assembled in a domain similarly to the α-α domain previously described for the protein prenyl transferase. The structures of the cis-type dimeric enzymes, undecaprenyl pyrophosphate synthase (UPPS) from E. coli and M. luteus, reveal two hydrophobic tunnels each surrounded by two α-helices and four β-strands. Both UPPS enzymes require Mg2+ for activity even though both lack the classical prenyl diphosphate Mg2+ binding motif (i.e., the (N/D)DXXD motif) found in most other trans-prenyltransferases and terpene synthases. The structures of terpenoid cyclases such as pentalene synthase, 5-epi-aristolochene synthase and trichodiene synthase harbor the similar structural feature referred to as “terpenoid synthase fold” with 10-12 mostly anti-parallel α-helices, as also observed in isoprenyl pyrophosphate synthases and protein prenyltransferases (see Liang, Eur J Biochem 269(14):3339-54 (2002)). All of the above cited structures differ greatly from the β/α-barrel fold described herein.
While a bound TAPS molecule in the first Orf2 structure tentatively indicated the approximate location of the diphosphate binding site near Asp 62 (FIG. 4A), structures complexed with GPP or a non-hydrolyzable analogue, GSPP, precisely define the residues involved in recognition and binding of the complete GPP substrate (FIGS. 4B and 4C). Lys 119, Asn 173 and Arg 228, located near the polar open end of the barrel, are hydrogen bonded to the terminal β-phosphate of the GSPP molecule (FIG. 4C). The α-phosphate linked to the geranyl chain hydrogen bonds with Tyr 216 and Lys 284, and also coordinates a Mg2+ ion. The complete coordination geometry of this Mg2+ ion exhibits perfect octahedral symmetry, with four equatorially arranged water molecules and two axially located oxygen atoms contributed by the side chain carboxylate of Asp 62 and an α-phosphate non-bridging oxygen of the GSPP molecule (FIGS. 4B and 4C). Despite the absence of a (N/D)DXXD motif, a second well conserved residue, Asp 110, proximal to Asp 62 in the tertiary structure, indirectly coordinates the Mg2+ ion via one of the four equatorially arranged water molecules. Tyr 121 resides within hydrogen bonding distance of the bridging atom (sulfur in GSPP and oxygen in GPP) linking the diphosphate moiety to the C10 geranyl chain. Finally, the hydrophobic geranyl chain of the GPP or GSPP molecules rest against the side chains of Val 49, Phe 123, Met 162, Tyr 175 and Tyr 216 (FIG. 4C).
The ternary complexes with Mg2+, GSPP and either 1,6-DHN or flaviolin delineate the chemical nature of the aromatic substrate binding site (FIGS. 4A, 4B and 4C). 1,6-DHN rests against the GSPP prenyl tail and is sequestered between the side chains of Met 162 and Phe 213. The Gln 295 and Leu 298 side chains provided by the short C-terminal helix line the wall of the substrate binding pocket with additional contacts made through the side chains of Phe 213, Ser 214 and Tyr 288. Flaviolin binds in a slightly different position than 1,6-DHN with extra pairs of hydrogen bonds formed with Ser 214, Tyr 288 and Gln 295, while the aromatic planes of both 1,6-DHN and flaviolin reside in the same active site orientation (FIG. 4A).
While not wishing to be bound by any theory, the structures of the substrates and products are consistent with an electrophilic aromatic substitution for the alkylation. Theoretically, two catalytic mechanisms can be considered for prenylation of aromatic substrates. One invokes a carbon mediated nucleophilic attack on the C1 carbon of GPP with the diphosphate moiety serving as a leaving group stabilized by Mg2+ coordination and the basic character of the diphosphate binding site. This Sn2-like mechanism has been described for protein farnesyltransferase (Park et al., supra; Long et al., supra). A second mechanism is reminiscent of terpene synthases involved in allylic diphosphate biosynthesis and prenyl group cyclization and invokes carbocation mediated electrophilic capture as proposed for the trans-prenyltransferase reaction of FPP synthase (Tarshis et al., Biochemistry 33(36):10871-7 (1994)) and numerous terpene synthases (cyclases) of secondary metabolism (Cane, in Comprehensive Naturals Products Chemistry: Isoprenoids, D. E. Cane, Editor, 1998, Elsevier Science: Oxford, UK).
The distance between the C5 atom of 1,6-DHN, which is the identified site for prenylation, or the C3 atom of flaviolin, and the C1 atom of GSPP are 4 Å and 7 Å, respectively. Notably, these distances are similar to the 7.3 Å separation recently described in human protein farnesyltransferase between the C1 atom of a bound farnesyl diphosphate (FPP) molecule and a Cys residue on a peptide substrate (Long, 2002 supra). Even though an Sn2-like mechanism has been proposed for prenyltransferases, these distances, combined with the structures of substrates and products, and the apparent requirement for a conformational change of the cleaved prenyl chain are consistent with an electrophilic aromatic substitution mechanism for Orf2-mediated alkylation of aromatic substrates (see FIG. 5).
A model for the overall reaction catalyzed by Orf2 with 1,6-DHN serving as the prenyl accepting group is depicted in FIG. 5. Firstly, a carbocation intermediate is proposed to result from the ionization of the diphosphate moiety, triggered by Mg2+ coordination, electrostatic hydrogen bonds with Lys 119, Arg 228, Asn 173 and Lys 284, and co-substrate binding. The positively charged C1 atom of the geranyl carbocation rotates toward the target double bond located 7 Å away on the prenyl acceptor (as previously described for human protein farnesyltransferase; see also Long, 2002, supra). A “tyrosine belt” including Tyr 121, Tyr 175 and Tyr 216, surrounding the 10 carbons of GPP, and similar to the one observed in the human protein farnesyltransferase (Park et al., supra; Long et al., Biochemistry 37(27):9612-8 (1998), may help stabilize and position the carbocationic intermediates via cation-π interactions (Wise and Croteau, in Comprehensive Naturals Products Chemistry: Isoprenoids, D. E. Cane, Editor, 1998, Elsevier Science: Oxford, UK).
This step involves the attachment of the reactive electrophile to the C5 atom of the 1,6-DHN molecule to form a resonance stabilized carbocation or σ-complex (Olah and Mo, J. Am. Chem. Soc. 94:9241 (1972)) (FIG. 5). Finally, Tyr 216, which interacts with the diphosphate moiety of the GPP molecule, is also hydrogen bonded to a conserved and well ordered network of water molecules linked to the diphosphate moiety and located just above the co-substrate binding location. One of these water molecules, highlighted in FIG. 5, is ideally positioned to abstract an acidic proton from the prenylated C5 atom of the cationic σ-complex allowing for the restoration of the neutral aromatic now containing a covalently tethered geranyl chain.
To confirm the enzymatic importance of certain active site residues, preliminary mutational studies of Orf2 were carried out, and residual activities were monitored using cell extracts containing mutant enzymes. The D62S and D62N single mutants, as well as D62S/S51R and D62N/S51K double mutants, displayed only residual activity in the presence of GPP (in the presence or absence of Mg2+), while no detectable activity was observed for the D62A single mutant with either GPP or DMAPP as a prenyl donor, indicative of the importance of D62 in catalytic processes.
In order to decipher the prenyl diphosphate chain length selectivity, molecular determinants of aromatic substrate recognition and divalent cation dependence, homology modeling of CloQ (Streptomyces roseochromogenes, accession number AF329398), NovQ (Streptomyces spheroides NCIMB 11891, accession number AF170880) and HypSc (Streptomyces coelicolor A3(2), accession number AL939130) sequences were carried out using the three dimensional architecture of Orf2 as a structural template (FIG. 6). The large degree of overall sequence similarity between these sequences as well as the considerable degree of active site conservation between Orf2 and CloQ/NovQ/HypSc is indicative of the conservation of the β/α-barrel fold for this family of aromatic prenyltransferases.
In the HypSc model, Asp 62 is replaced by an Asn residue and is complemented by the replacement of Ser 52 by an Arg residue. Also, as observed in the CloQ/NovQ model, the presence of a salt bridge between Arg 65 and Glu 278 appears from modeling to prevent the binding of a prenyl donor with an alkyl chain longer thatn C5 (i.e., C10 or C15). From this modeling analysis, the deduced protein from Streptomyces coelicolor would be predicted to show a DMAPP specificity and Mg2+ independent prenyltransferase activity.
In order to validate this model, HypSc was subcloned from genomic DNA, and over-expressed in E. coli as an octa-histidine tagged protein and purified for Ni2+-chelation chromatography. The purified enzyme was then assayed for prenyltransferase activity using DMAPP and GPP as prenyl donors. Notable prenyltransferase activity was detected when using DMAPP and 1,6-DHN as substrates in the absence of Mg2+, consistent with the model based hypothesis set forth herein regarding the chain length selectivity and Mg2+ independence of invention enzymes.
In accordance with yet another aspect of the present invention, there are provided compositions comprising an aromatic prenyltransferase as described herein in crystalline form. Optionally, such compositions further comprise one or more substrates for the aromatic prenyltransferase. As used herein, “substrates” refer to compounds susceptible to the action of invention prenyltransferases, e.g., such reactive aromatic compounds as tetrahydroxynaphthalene, analogs, homologs and metabolites thereof.
As used herein, “analogs” refer to compounds which are related to the above-described aromatic substrates and retain a biological activity thereof, but have one or more substitutions and/or modifications thereof relative to the parent compound, e.g., substitution of —O— for —CH2—. Alternatively, analogs may have relatively little primary structure similarity, but may still display a biological activity of a substrate as a result of similar secondary and/or tertiary structural features, electronic properties, and the like.
As used herein, “homolog” refers to compounds which are related to the above-described aromatic substrates by the presence or absence of a simple unit, such as a methylene unit, or some multiple of such units, e.g., —(CH2)x—.
As used herein, “metabolite” refers to compounds which are related to the above-described substrates as a form of such compound obtained in a human or animal body by action of the body on the administered form of the compound, for example a de-methylated analogue of a compound bearing a methyl group which is obtained in the body after administration of the methylated compound as a result of action by the body on the methylated compound.
X-ray crystallography can elucidate the three-dimensional structure of crystalline forms according to the invention. Typically, the first characterization of crystalline forms by X-ray crystallography can determine the unit cell shape and its orientation in the crystal. The term “unit cell” refers to the smallest and simplest volume element of a crystal that is completely representative of the unit of pattern of the crystal. The dimensions of the unit cell are defined by six numbers: dimensions a, b and c and angles α, β and γ. A crystal can be viewed as an efficiently packed array of multiple unit cells. Detailed descriptions of crystallographic terms are provided in Hahn, THE INTERNATIONAL TABLES FOR CRYSTALLOGRAPHY, VOLUME A, 4th Ed., Kluwer Academic Publishers (1996); and Shmueli, THE INTERNATIONAL TABLES FOR CRYSTALLOGRAPHY, VOLUME B, 1st Ed., Kluwer Academic Publishers. The term “space group” refers to the symmetry of a unit cell. In a space group designation (e.g., P2) the capital letter indicates the lattice type and the other symbols represent symmetry operations that can be carried out on the unit cell without changing its appearance.
The term “selenomethionine substitution” refers to the method of producing a chemically modified form of a protein crystal. The protein is expressed by bacteria in media that is depleted in methionine and supplemented with selenomethionine. Selenium is thereby incorporated into the crystal in place of methionine sul furs. The location(s) of selenium is(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 a protein 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 and Johnson, PROTEIN CRYSTALLOGRAPHY, Academic Press (1976), which is incorporated by reference herein.
The knowledge obtained from X-ray diffraction patterns can be used in the determination of the three-dimensional structure of the binding sites of other homologous polypeptides. This is achieved through the use of commercially available software known in the art that is capable of generating three-dimensional graphical representations of molecules or portions thereof from a set of structure coordinates. The binding domain can also be predicted by various computer models. Based on the structural X-ray coordinates of the solved structure, mutations and variants of the solved structure can also be designed.
An exemplary isolated aromatic prenyltransferase according to the present invention has been further characterized by the structural coordinates set forth in Appendix 1.
In accordance with still another aspect of the present invention, there are provided methods of predicting the activity and/or substrate specificity of a putative aromatic prenyltransferase, the methods comprising:
comparing a three-dimensional representation of a known aromatic prenyltransferase and a three-dimensional representation of a putative aromatic prenyltransferase, wherein differences between the two representations are predictive of aromatic prenyltransferase activity and/or substrate specificity.
In accordance with yet another aspect of the present invention, there are provided methods of screening for compounds which bind aromatic prenyltransferase(s), said methods comprising:
modeling a potential binding agent that interacts with one or more domains of an aromatic prenyltransferase or fragment thereof, defined by a plurality of atomic coordinates of the aromatic prenyltransferase or fragment thereof, and
determining the ability of said potential binding agent to compete with said aromatic prenyltransferase substrate for binding to said aromatic prenyltransferase.
As used herein, “molecular replacement” refers to generating a preliminary model of a polypeptide whose structure coordinates are unknown, by orienting and positioning a molecule whose structure coordinates are known within the unit cell of the unknown crystal so as to best 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, Meth. Enzymol. 115:55-77 (1985); Rossmann, M G., ed., THE MOLECULAR REPLACEMENT METHOD (1972), Int. Sci. Rev. Ser. No. 13, Gordon & Breach, New York). Using structure coordinates of the aromatic prenyltransferase provided herein, molecular replacement may be used to determine the structure coordinates of a crystalline mutant, homologue, or a different crystal form of an aromatic prenyltransferase.
In accordance with this invention, an aromatic prenyltransferase, or a portion thereof, may be crystallized in association or complex with any known or putative substrate and/or binding agent. The crystal structures of a series of such complexes may then be solved by molecular replacement and compared with that of a native aromatic prenyltransferase molecule. Potential sites for modification within the aromatic prenyltransferase molecule or a corresponding substrate and/or binding agent therefor may thus be identified based on the points of interaction between the aromatic prenyltransferase and substrate and/or binding agent therefor. This information provides an additional tool for determining the most efficient binding interactions, for example, increased hydrophobic interactions, between an aromatic prenyltransferase and a putative chemical entity or compound, even before any synthesis or modifications are performed.
All of the complexes referred to above may be studied using well-known X-ray diffraction techniques as described herein, and may be refined versus 2-3 Å resolution X-ray data to an R value of about 0.20 or less using computer software, such as X-PLOR (Yale University, 1992, distributed by Molecular Simulations, Inc.). See, e.g., Blundel & Johnson, supra; Methods in Enzymology, vol. 114 and 115, H. W. Wyckoffet al., eds., Academic Press (1985). This information may thus be used to optimize known classes of aromatic prenyltransferase substrate and/or binding agent therefor, such as natural THN, and to design, modify and/or synthesize novel classes of aromatic prenyltransferase substrate and/or binding agents.
The modeling or design of substrates and/or binding agents for aromatic prenyltransferases, i.e., compounds that bind to and/or modulate an aromatic prenyltransferase polypeptide according to the invention generally involves consideration of two factors. First, the compound or molecule must be capable of physically and structurally associating with an aromatic prenyltransferase molecule. Non-covalent molecular interactions important in the association of an aromatic prenyltransferase with a putative substrate and/or binding agent include hydrogen bonding, van der Waals and hydrophobic interactions, and the like.
Second, the compound or molecule must be able to assume a conformation that allows it to associate with an aromatic prenyltransferase molecule. Although certain portions of the compound or molecule 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 affinity with the receptor. Such conformational requirements include the overall three-dimensional structure and orientation of the compound or molecule in relation to all or a portion of the binding site, or the spacing between functional groups of a compound or molecule comprising several chemical entities that directly interact with an aromatic prenyltransferase.
The term “modeling” as used herein, refers to analysis of the interaction of an aromatic prenyltransferase and a known or test compound or molecule by utilizing a computer generated representation of the molecules, as opposed to physical molecules.
The potential binding of a test compound with an aromatic prenyltransferase may be analyzed prior to its actual synthesis and testing by the use of computer modeling techniques. If the theoretical structure of the given compound is indicative of insufficient interaction and association between it and an aromatic prenyltransferase, synthesis and testing of the compound may be obviated. However, if computer modeling indicates a strong interaction, the molecule may then be tested for its ability to bind to an aromatic prenyltransferase. Methods of assaying for aromatic prenyltransferase activity are known in the art (as identified and discussed herein). Methods for assaying the effect of a potential binding agent can be performed in the presence of a known binding agent of an aromatic prenyltransferase. 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 binding agent.
A test compound 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 an aromatic prenyltransferase associated with a substrate and/or binding agent therefor.
One skilled in the art may use one of several methods to predict a molecule capable of binding to an aromatic prenyltransferase and to screen test compounds for their ability to associate with an aromatic prenyltransferase and more particularly with the individual active site (e.g., binding pockets and/or specific points of interaction) of an aromatic prenyltransferase polypeptide. This process may begin by visual inspection of, for example, the binding pocket of an aromatic prenyltransferase on the computer screen based on structure coordinates obtained derived from X-ray diffraction data obtained from crystals of an aromatic prenyltransferase, such as those provided in Appendix 1. Selected fragments or chemical entities may then be positioned in a variety of orientations, or docked, within an individual binding pocket of the aromatic prenyltransferase. 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 at this stage. 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 chemical entities or fragments have been selected, they can be assembled into a single compound that is a candidate substrate and/or binding agent. 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 aromatic prenyltransferase molecule as set forth in Appendix 1. 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 a substrate and/or binding agent in a step-wise fashion one fragment or chemical entity at a time as described above, aromatic prenyltransferase substrates and/or binding agents may be designed as a whole or “de novo” using either an empty binding pocket or optionally including some portion(s) of a known substrate(s) and/or binding agent(s). 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 test compound or binding agent has been designed or selected by the above methods, the efficiency with which that compound may bind to an aromatic prenyltransferase may be tested and optimized by computational evaluation.
A compound designed or selected as a putative substrate and/or 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 an aromatic prenyltransferase when the substrate and/or binding agent is bound to the aromatic prenyltransferase, 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.
Other molecular modeling techniques may also be employed in accordance with this invention. For exemplary reviews and techniques, see, e.g., Cohen et al., “Molecular Modeling Software and Methods for Medicinal Chemistry, J. Med. Chem., 33, pp. 883-894 (1990); see also, M. A. Navia and M. A. Murcko, “The Use of Structural Information in Drug Design”, Current Opinions in Structural Biology, 2, pp. 202-210 (1992); L. M. Balbes et al., “A Perspective of Modern Methods in Computer-Aided Drug Design”, in Reviews in Computational Chemistry, Vol. 5, K. B. Lipkowitz and D. B. Boyd, Eds., VCH, New York, pp. 337-380 (1994); see also, W. C. Guida, “Software For Structure-Based Drug Design”, Curr. Opin. Struct. Biology, 4, pp. 777-781 (1994)]
In accordance with still another aspect of the present invention, there are provided alternate methods of screening for compounds which bind aromatic prenyltransferase(s), said methods comprising:

    
    
        defining an interaction site of an aromatic prenyltransferase based on a plurality of atomic coordinates of said aromatic prenyltransferase;
        modeling a potential binding agent that fits spatially into said interaction site;
        contacting said potential binding agent with said aromatic prenyltransferase in the presence of an aromatic prenyltransferase substrate; and
        determining the ability of said potential binding agent to compete with said aromatic prenyltransferase substrate for binding to said aromatic prenyltransferase.
    
    


In accordance with a further aspect of the present invention, there are provided additional methods of screening for compounds which bind aromatic prenyltransferase(s), said methods comprising:

    
    
        defining an interaction site of an aromatic prenyltransferase based on a plurality of atomic coordinates of said aromatic prenyltransferase;
        modeling a potential binding agent that fits spatially into said interaction site; and
        determining the ability of said potential binding agent to compete with an aromatic prenyltransferase substrate for said interaction site by contacting said potential binding agent with said aromatic prenyltransferase in the presence of said aromatic prenyltransferase substrate.
    
    


In accordance with a still further aspect of the present invention, there are provided additional methods of screening for compounds which bind aromatic prenyltransferase(s), said methods comprising:

    
    
        modeling a potential binding agent that fits spatially into an interaction site of an aromatic prenyltransferase defined by a plurality of atomic coordinates of said aromatic prenyltransferase;
        contacting said potential binding agent with said aromatic prenyltransferase in the presence of an aromatic prenyltransferase substrate; and
        determining the ability of said potential binding agent to compete with said aromatic prenyltransferase substrate for binding to said aromatic prenyltransferase.
    
    


In accordance with another aspect of the present invention, there are provided additional methods of screening for compounds which bind aromatic prenyltransferase(s), said methods comprising:

    
    
        modeling a potential binding agent that fits spatially into an interaction site of an aromatic prenyltransferase defined by a plurality of atomic coordinates of said aromatic prenyltransferase; and
        determining the ability of said potential binding agent to compete with an aromatic prenyltransferase substrate for said interaction site by contacting said potential binding agent with said aromatic prenyltransferase in the presence of said aromatic prenyltransferase substrate.
    
    


In accordance with yet another aspect of the present invention, there are provided additional methods of screening for compounds which bind aromatic prenyltransferase(s), said methods comprising:

    
    
        determining the ability of a potential binding agent to compete with an aromatic prenyltransferase substrate for binding to an aromatic prenyltransferase, wherein the potential binding agent is modeled to fit spatially into an aromatic prenyltransferase interaction site defined by a plurality of atomic coordinates.
    
    


In accordance with still another aspect of the present invention, there are provided methods of identifying potential substrate(s) of an aromatic prenyltransferase, said methods comprising:

    
    
        defining an active site of said aromatic prenyltransferase based on a plurality of atomic coordinates of said aromatic prenyltransferase;
        identifying a potential substrate that fits said active site; and
        contacting the aromatic prenyltransferase with the potential substrate and determining its activity thereon.
    
    


In accordance with a further aspect of the present invention, there are provided methods of screening compounds to determine whether such compounds are aromatic prenyltransferase substrates, said methods comprising:

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


In accordance with still another aspect of the present invention, there are provided alternate methods of screening compounds to determine whether such compounds are aromatic prenyltransferase substrates, said methods comprising:

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


In accordance with a still further aspect of the present invention, there are provided additional methods of screening compounds to determine whether such compounds are aromatic prenyltransferase substrates, said methods comprising:

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


In accordance with yet another aspect of the present invention there are provided methods for stimulating the activity of an aromatic prenyltransferase, said methods comprising contacting said aromatic prenyltransferase with an effective amount of a compound identified by any of the above-described methods.
Such compounds are typically administered as part of formulations comprising at least one of the above-described compounds in a pharmaceutically acceptable carrier therefor. Exemplary pharmaceutically acceptable carriers include solids, solutions, emulsions, dispersions, micelles, liposomes, and the like. Optionally, the pharmaceutically acceptable carrier employed herein further comprises an enteric coating.
Pharmaceutically acceptable carriers contemplated for use in the practice of the present invention are those which render invention compounds amenable to oral delivery, transdermal delivery, intravenous delivery, intramuscular delivery, topical delivery, nasal delivery, and the like.
Thus, formulations contemplated for use in the practice of the present invention can be used in the form of a solid, a solution, an emulsion, a dispersion, a micelle, a liposome, and the like, wherein the resulting formulation contains one or more of the compounds of the present invention, as an active ingredient, in admixture with an organic or inorganic carrier or excipient suitable for enterable or parenteral applications. The active ingredient may be compounded, for example, with the usual non-toxic, pharmaceutically acceptable carriers for tablets, pellets, capsules, suppositories, solutions, emulsions, suspensions and any other suitable for use. The carriers which can be used include glucose, lactose, gum acacia, gelatin, manitol, starch paste, magnesium trisilicate, talc, corn starch, keratin, colloidal silica, potato starch, urea, medium chain length triglycerides, dextrans, and other carriers suitable for use in manufacturing preparations, in solid, semisolid, or liquid form. In addition auxiliary, stabilizing, thickening, and coloring agents and perfumes may be used. The active compound(s) is (are) included in the formulation in an amount sufficient to produce the desired effect upon the process or disease condition.
Formulations contemplated for use in the practice of the present invention containing the active ingredient may be in a form suitable for oral use, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Formulations intended for oral use may be prepared according to any method known to the art for the manufacture of pharmaceutical compositions and such formulations may contain one or more agents selected from the group consisting of a sweetening agent such as sucrose, lactose, or saccharin, flavoring agents such as peppermint, oil of wintergreen or cherry, coloring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets containing the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients used may be, for example (1) inert diluents such as calcium carbonate, lactose, calcium phosphate or sodium phosphate; (2) granulating and disintegrating agents such corn starch, potato starch or alginic acid; (3) binding agents such as gum tragacanth, corn starch, gelatin or acacia, and (4) lubricating agents such as magnesium stearate, steric acid or talc. The tablets may be uncoated or they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate may be employed. They may also be coated by such techniques as those described in U.S. Pat. Nos. 4,256,108; 4,160,452; and 4,265,874, to form osmotic therapeutic tablets for controlled release.
In some cases, formulations contemplated for oral use may be in the form of hard gelatin capsules wherein the active ingredient is mixed with inert solid diluent(s), for example, calcium carbonate, calcium phosphate or kaolin. They may also be in the form of soft gelatin capsules wherein the active ingredient is mixed with water or an oil medium, for example, peanut oil, liquid paraffin, or olive oil.
Formulations contemplated for use in the practice of the present invention may be in the form of a sterile injectable suspension. This suspension may be formulated according to known methods using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally-acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil may be employed including synthetic mono- or diglycerides, fatty acids, naturally occurring vegetable oils like sesame oil, coconut oil, peanut oil, cottonseed oil, etc. or synthetic fatty vehicles like ethyl oleate or the like. Buffers, preservatives, antioxidants, and the like can be incorporated as required.
Formulations contemplated for use in the practice of the present invention may also be administered in the form of suppositories for rectal administration of the drug. These formulations may be prepared by mixing the drug with a suitable non-irritating excipient, such as cocoa butter, synthetic glyceride esters of polyethylene glycols, which are solid at ordinary temperatures, but liquefy and/or dissolve in the rectal cavity to release the drug. Since individual subjects may present a wide variation in severity of symptoms and each drug has its unique therapeutic characteristics, the precise mode of administration and dosage employed for each subject is left to the discretion of the practitioner.
Amounts effective for the particular therapeutic goal sought will, of course, depend on the severity of the condition being treated, and the weight and general state of the subject. Various general considerations taken into account in determining the “effective amount” are known to those of skill in the art and are described, e.g., in Gilman et al., eds., Goodman And Gilman's: The Pharmacological Bases of Therapeutics, 8th ed., Pergamon Press, 1990; and Remington's Pharmaceutical Sciences, 17th ed., Mack Publishing Co., Easton, Pa., 1990, each of which is herein incorporated by reference.
The term “effective amount” as applied to compounds contemplated for use in the practice of the present invenion, means the quantity necessary to effect the desired therapeutic result, for example, a level effective to treat, cure, or alleviate the symptoms of a disease state for which the therapeutic compound is being administered, or to establish homeostasis. Since individual subjects may present a wide variation in severity of symptoms and each drug or active agent has its unique therapeutic characteristics, the precise mode of administration, dosage employed and treatment protocol for each subject is left to the discretion of the practitioner.
The above-described methods for stimulating activity of aromatic prenyltransferases can be applied in many situations. For example, cancer cell resistance to chemotherapy is often mediated by overexpression of P-glycoprotein, a plasma membrane ABC (ATP-binding cassette) transporter which extrudes cytotoxic drugs at the expense of ATP hydrolysis. Prenylated flavoinoids have recently been reported as potential inhibitor of human multidrug resistant protein (MRP1) which belong to the ABC transporter superfamilly. Some of these prenylated compounds have also shown HIV-inhibitory effects. Recently, common forms of the prenylated flavonoids have been identified in beer: 6- and 8-prenylnaringenin, xanthohumol and isoxanthohumol are present in high concentrations in hops (Humulus lupulus L.) and their oestrogenic potency has been determined in in vitro and animal model systems, with data indicating that they are more potent oestrogens than the isoflavones class (see Milligan et al., J Clin Endocrinol Metab 84:2249-52 (1999).
In accordance with a still further aspect of the present invention, there are provided methods of identifying potential modulator(s) of aromatic prenyltransferase(s), said methods comprising:

    
    
        defining an aromatic prenyltransferase polypeptide or fiagment thereof based on a plurality of atomic coordinates of the aromatic prenyltransferase polypeptide;
        modeling a potential binding agent that interacts with one or more domains of the aromatic prenyltransferase polypeptide;
        contacting the potential binding agent with the aromatic prenyltransferase polypeptide; and
        determining the ability of said potential binding agent to modulate an aromatic prenyltransferase biological function, thereby identifying a potential modulator of an aromatic prenyltransferase polypeptide.
    
    


As employed herein, “modulators” refers to compound(s) which, either directly (by binding to a prenyltransferase) or indirectly (as a precursor for a compound which binds to a prenyltransferase, or an inducer which promotes production of a compound which binds to a prenyltransferase from a precursor) induce the activity of prenyltransferase, or to repress the activity of prenyltransferase. Exemplary modulators contemplated in the practice of the present invention include flavonoids, isoflavonoids, and the like.
In accordance with yet another aspect of the present invention, there are provided alternate methods of identifying potential modulator(s) of the activity of aromatic prenyltransferase(s), said methods comprising:

    
    
        defining the active site of said aromatic prenyltransferase based on a plurality of atomic coordinates of said aromatic prenyltransferase,
        contacting a potential compound that fits the active site of (a) with the aromatic prenyltransferase in the presence of a substrate, and
        determining the ability of said compound to modulate the activity of said aromatic prenyltransferase with respect to said substrate.
    
    


In accordance with still another aspect of the present invention, there are provided additional methods of identifying potential modulator(s) of the activity of aromatic prenyltransferase(s), said methods comprising:

    
    
        contacting a potential compound that fits an active site based on a plurality of atomic coordinates of said aromatic prenyltransferase; and
        determining the ability of said compound to modulate the activity of said aromatic prenyltransferase.
    
    


In accordance with a further aspect of the present invention, there are provided methods of screening for compounds that modulate the activity of aromatic prenyltransferase(s), said methods comprising:

    
    
        determining the points of interaction between an aromatic prenyltransferase, and substrate or substrate mimic therefor;
        selecting compound(s) having similar interaction with said aromatic prenyltransferase; and
        testing the selected compound for the ability to modulate the activity of an aromatic prenyltransferase.
    
    


As employed herein, “modulating” refers to the ability of a modulator for a prenyltransferase to either directly or indirectly induce prenyltransferase activity, or to repress prenyltransferase activity. Exemplary processes contemplated for modulation according to the invention include cholesterol metabolism, regulation of lipid homeostasis, stimulation of bile transport and absorption, regulation of the expression of genes involved in the excretion and transportation of bile acids (including intestinal bile acid-binding protein (IBABP)), bile salt export pump (BSEP) and canalicular multi-specific organic anion transporter (cMOAT), and the like.
In accordance with still another aspect of the present invention, there are provided alternate methods of screening for compounds that modulate the activity of aromatic prenyltransferase(s), said methods comprising:

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


In accordance with yet another aspect of the present invention, there are provided additional methods of screening for compounds that modulate the activity of aromatic prenyltransferase(s), said methods comprising:

    
    
        testing a compound for the ability to modulate the activity of an aromatic prenyltransferase,
        wherein said compound has been selected as having points of interaction with said aromatic prenyltransferase, and
        wherein similar points of interaction have been determined between said aromatic prenyltransferase and a substrate or substrate mimic therefor.
    
    


In accordance with yet another aspect of the present invention, there are provided methods for prenylating aromatic substrates, said methods comprising:

    
    
        contacting an aromatic substrate with an aromatic prenyltransferase as described herein, under prenylating conditions.
    
    


In accordance with still another aspect of the present invention, there are provided methods of identifying proteins having a beta/alpha barrel structure, said methods comprising:

    
    
        comparing a three-dimensional representation of an aromatic prenyltransferase as described herein with a three-dimensional representation of a putative protein having a beta/alpha barrel structure, wherein similarities between the two representations are predictive of aromatic prenyltransferase proteins having a beta/alpha barrel structure.
    
    


In accordance with a further aspect of the present invention, there are provided methods for controlling the degree of prenylation promoted by an aromatic prenyltransferase, said methods comprising:

    
    
        altering one or more active site residues of said aromatic prenyltransferase so as to change the dimensions of the active site sufficiently to control the degree of prenylation promoted by said aromatic prenyltransferase.
    
    


As used herein, “degree of prenylation” refers to the number of isoprenoid units added to a substrate. This embraces prenylation at multiple sites, as well as introduction of one or more isoprenoid units at a single site.
In accordance with a still further aspect of the present invention, there are provided methods for modifying the degree of prenylation promoted by an aromatic prenyltransferase, said methods comprising:

    
    
        modifying one or more active site residues of said aromatic prenyltransferase so as to change the dimensions of the active site sufficiently to modify the degree of prenylation promoted by said aromatic prenyltransferase.
    
    


In accordance with yet another aspect of the present invention, there are provided methods for controlling the substrate specificity of an aromatic prenyltransferase, said methods comprising:

    
    
        altering one or more active site residues of said aromatic prenyltransferase so as to change the dimensions of the active site sufficiently to control the selectivity of said aromatic prenyltransferase with respect to aromatic substrates which are prenylated by said aromatic prenyltransferase.
    
    


As used herein, “substrate specificity” refers to the selectivity with which an enzyme recognizes a substrate. A selective prenyltransferase will recognize only a single, or a limited number of substrates, whereas a non-selective (promiscuous) prenyltransferase will recognize a plurality of substrates.
In accordance with a further aspect of the present invention, there are provided methods for modifying the substrate specificity of an aromatic prenyltransferase, said methods comprising:

    
    
        modifying one or more active site residues of said aromatic prenyltransferase so as to change the dimensions of the active site sufficiently to modify the selectivity of said aromatic prenyltransferase with respect to aromatic substrates which are prenylated by said aromatic prenyltransferase.
    
    


In accordance with still another aspect of the present invention, there are provided methods for controlling the donor specificity of an aromatic prenyltransferase, said methods comprising:

    
    
        altering one or more active site residues of said aromatic prenyltransferase so as to change the dimensions of the active site sufficiently to control the selectivity of said aromatic prenyltransferase with respect to prenyl donors which are employed to prenylate an aromatic substrate.
    
    


As used herein, “donor specificity” refers to the selectivity with which an enzyme recognizes a prenyl donor. A selective prenyltransferase will recognize only a single, or a limited number of prenyl donors, whereas a non-selective (promiscuous) prenyltransferase will recognize a plurality of prenyl donors. Exemplary prenyl donors include dimethylallyl diphosphate (DMAPP, C5), isopentenyl diphosphate (IPP, C5), geranyl diphosphate (GPP, C10), farnesyl diphosphate (FPP, C15), and the like.
In accordance with a further aspect of the present invention, there are provided methods for modifying the donor specificity of an aromatic prenyltransferase, said methods comprising:

    
    
        modifying one or more active site residues of said aromatic prenyltransferase so as to change the dimensions of the active site sufficiently to modify the selectivity of said aromatic prenyltransferase with respect to prenyl donors employed to prenylate an aromatic substrate.
    
    


In accordance with still another aspect of the present invention, there are provided computer programs on a computer readable medium, said computer programs comprising instructions to cause a computer to define an aromatic prenyltransferase or fragment thereof based on a plurality of atomic coordinates of the aromatic prenyltransferase.
According to another aspect of the present invention, there is provided a computer for determining at least a portion of the structure coordinates corresponding to X-ray diffraction data obtained from an aromatic prenyl transferase molecule or molecular complex or a homologue of said aromatic prenyl transferase molecule or molecular complex, said computer comprising:

    
    
        (i) a computer-readable data storage medium comprising a data storage material encoded with machine-readable data, wherein said data comprises at least a portion of the structure coordinates of Appendix 1;
        (ii) a computer-readable data storage medium comprising a data storage material encoded with computer-readable data, wherein said data comprises X-ray diffraction data obtained from said aromatic prenyl transferase molecule or molecular complex or a homologue of said aromatic prenyl transferase molecule or molecular complex;
        (iii) a working memory for storing instructions for processing said computer-readable data of (i) and (ii);
        (iv) a central-processing unit coupled to said working memory and to said computer-readable data storage medium of (i) and (ii) for performing a Fourier transform of the machine readable data of (i) and for processing said computer-readable data of (ii) into structure coordinates; and
        (v) a display coupled to said central-processing unit for displaying said structure coordinates of said molecule or molecular complex.
    
    


The term “computer” as used herein can be composed of a central processing unit (for example, the Pentium III from Intel Corporation, or similar processor from Sun, Motorola, Compaq, AMD or International Business Machines, and the like), a working memory which may be random-access memory or core memory, mass storage memory (for example, one or more floppy disk drives, compact disk drives or magnetic tape containing data recorded thereon), at least one display terminal, at least one keyboard and accompanying input and output devices and connections therefor. The computer typically includes a mechanism for processing, accessing and manipulating input data. A skilled artisan can readily appreciate that any one of the currently available computer systems are suitable. It should also be noted that the computer can be linked to other computer systems in a network or wide area network to provide centralized access to the information contained within the computer.
Contemplated input devices for entering machine readable data include, for example, telephone modem lines, cable modems, CD-ROMs, a keyboard or disk drives. The computer may advantageously include or be programmed with appropriate software for reading the data from the data storage component or input device, for example computational programs for use in rational drug design that are described in detail below. Contemplated output devices include conventional systems known in the art, for example, display terminals, printers, or disk drives for further storage of output.
Embodiments of the invention include systems (e.g., internet 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. 8. As used herein, “a computer system” refers to the hardware components, software components, and data storage components used to analyze the coordinates and sequences such as those set forth in Appendix 1. 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 other suppliers such as 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(s) 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, a modem capable of connection to a remote data storage system (e.g., via the internet), and the like. 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, and the like, 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 of Appendix 1, (such as search tools, compare tools, and modeling tools etc.) may reside in main memory 115 during execution (Appendix 1, discloses residues 3-303 of SEQ ID NO: 2).
Computer programs are widely available that are capable of carrying out the activities necessary to model structures and substrates using the crystal structure information provided herein. Examples include, but are not limited to, the computer programs listed below:

    
    
        CATALYST DATABASETM—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 an 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 this 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.
The following examples are provided to further illustrate aspects of the invention. These examples are non-limiting and should not be construed as limiting any aspect of the invention.
EXAMPLES
All solvents and reagents were obtained from the Aldrich Chemical Company (Milwaukee, Wis.) unless otherwise indicated.
Example 1
Cloning of ORF2
A cosmid pCLC7 (see Takagi et al., J Bacteriol 182:41534157 (2000)), which contains the mevalonate pathway gene cluster and the flanking regions cloned from CL190, was sequenced. The DNA sequence of Orf2 was determined by standard techniques, and is set forth as SEQ ID NO:1. The amino acid sequence of Orf2 was deduced from the DNA sequence and is set forth as SEQ ID NO:2.
This sequencing revealed 3 new complete orfs, orf1, orf2, and orf3 and a partial orf4 in a 9.0 kb-BamHI-BamHI DNA fragment which contains mevalonate kinase and diphosphomevalonate decarboxylase (see pCL3301 in FIG. 6). To deduce function of each orf, a database search was done. The results are summarized in Table 2.











TABLE 2









Most homologous proteins and



ORFs
Amino acids
their accession numbers









ORF1
319 aa
S. avermitilis RNA polymerase ECF-





subfamily σ factor, AP005050



ORF2
307 aa
S. coelicolor A3(2) protein,





AL391041



ORF3
410 aa
S. antibioticus type III





polyketide synthase, AB084489



ORF4
177 aa
S. erythraeus protein, AY078067










ORF2 also showed sequence similarity to the previously described 4-hydroxyphenylpyruvate:dimethylallytransferase, cloQ (accession number AF329398) and novQ (accession number AF 170880), from Steptomyces roseochromogenes and Streptomyces spheroides NCIMB 11891, respectively (Pojer et al., supra). ORF3 is most likely to encode type III polyketide synthase which produces THN. These data confirm that ORF2 encodes geranyltransferase which catalyzes geranyl transfer to THN or a THN derivative produced by the action of ORF3.
Example 2
Cloning, expression and purification of the ORF2 of SEQ ID NO:1 gene
The orf2 gene of SEQ ID NO:1 from Streptomyces sp. strain CL190 (GenBank accession number AB187169) was cloned by PCR amplification of total genomic DNA from CL190 using oligonucleotides designed for ligation into the E. coli expression vector pQE30 (QIAGEN), to generate the expression plasmid pQEORF2. PCR amplification of pQEORF2, using oligonucleotides designed for ligation into the E. coli expression vector pHlS8 (Jez et al., Biochemistry 39(5): 890-902 (2000)) was carried out using the forward primer sequence:
5′- GGG GGG GGA TCC TCC GAA GCC GCT GAT GTC G-3′ (SEQ ID NO:3; BamHI site underlined), and the reverse primer sequence:


    
    
        5′-GGG GGG GAA TTC TCA GTC CTC CAG CGA GTC G-3′ (SEQ ID NO:4; EcoRI site underlined) to generate the expression vector pHIS8ORF2. Constructs of pHIS8ORF2 were transformed into E. coli BL21 (DE3) from NOVOGEN. Recombinant Orf2 protein of SEQ ID NO:2 was obtained and purified using a standard protocol described before Jez et al., supra. Selenomethionine (Se-Met)-subsituted protein was obtained from E. coli grown in M9 minimal medium using the methionine pathway inhibition approach (Doublir, 1997), and purified as described for the native protein.
    
    


Example 3 
Crystallization of Orf2 of SEQ ID NO:2
were obtained by the vapor diffusion method at 40C. 2 μl hanging drops containing a 1:1 mixture of a 15 mg/ml protein with crystallization buffer (28% [w/v] PEG 4000, 0.3 M magnesium nitrate, 2 mM DL-dithiothreitol (DTT), 0.1 M PIPES pH 8.5) equilibrated over a 500 μl reservoir of the same solution produced small diffracting crystals overnight. Larger crystals were obtained by the macro-seeding technique in the same conditions. Crystals were stabilized by soaking briefly in a cryoprotectant solution (30% (w/v) PEG 4000, 15% (v/v) glycerol, 0.3 M magnesium nitrate, 2 mM DTT, 0.1 M TAPS, pH 8.5), and flash frozen in liquid nitrogen prior to data collection. Orf2 of SEQ ID NO:2 crystals belong to the P21212 space group with average unit cell dimensions of a=71, b=92 Å, c=48 Å, α=β=γ=90°., and contain one monomer per asymmetric unit and a solvent content of 45%. Se-Met substituted crystals were obtained as described (Doublie, Methods Enzymol 276:523-30 (1997)). Various complexes were obtained by soaking wild type Orf2 of SEQ ID NO:2 crystals in stabilization solution containing 5 mM GPP, 10 mM GSPP and 40 mM 1,6-DHN, and 10 mM GSPP and 10 mM flaviolin (GPP and GSPP were purchased from Echelon Biosciences Inc.).
Example 4
Structure Determination and Refinement
A multi-wavelength anomalous dispersion (MAD) data set was collected at the selenium edge on a Se-Met incorporated protein crystal at the Brookhaven National Laboratory (BNL) on beam line X8C. Data were processed with HKL2000 (Otwinowski and Minor, Methods Enzymol 307-326 (1997)), and reduced to a unique set of indexed intensities to a resolution of 1.6 Å. Single wavelength data sets were collected in house, at Brookhaven National Laboratory (BNL), the European Synchrotron Facility (ESRF), and at the Stanford Synchrotron Radiation Laboratory (SSRL) on the various complexes (Table 1). Phasing, density modification and automatic model building were carried out with the program suite Solve/Resolve (Terwilliger, Acta Crystallogr D Biol Crystallogr 58(Pt 11): 1937-40 (2002); Terwilliger and Berendzen, Acta Crystallogr D Biol Crystallogr 55(Pt 4):849-61 (1999)) providing a high quality initial electron density map, using 7 identified Se sites. Additional rounds of building and refinement were carried out with the programs O (Jones, The O Manual, 1993, Upsalla, Sweden) and CNS (Brünger, Acta Crystallogr D Biol Crystallogr 54(Pt 5):905-21 (1998)), respectively. This first model was used to solve the various complexes by molecular replacement with AMoRe (Navaza, Acta Crystallogr D Biol Crystallogr 57(Pt 10):1367-72 (2001)).
Example 5
Detection of prenyltransferase activity of ORF2 ofSEQ ID NO:2
The basal reaction buffer employed contained 50 mM HEPES (pH 7.5), 5 mM MgCl2, 5 mM DTT (as needed), 5 mM prenyl acceptor (DHN1, DHN2 and DHN3; see FIG. 2B), and optionally 5 mM FPP or GPP, in a final volume of 20 μl. The reaction (except for control) was initiated by adding 20 μg of ORF2 protein of SEQ ID NO:2 to the basal assay mixture. After incubation at room temperature for 4 hrs., the reaction mixture was dried using SPEEDVAC and the dried residue was spotted on a silica gel TLC plate. The plate was developed with chloroform:methanol (15-30:1). Reaction products were detected at 254 nm UV.
With prenyl acceptors DHN1 and DHN2, and either prenyl donor, FPP or GPP, prenyltransferase activity was observed. With prenyl acceptor DHN3, prenyltransferase activity was observed with GPP.
Additional studies were carried out with ORF2 and (a) 1,6-DHN (2), (b) 2,7-DHN (3), (c) daidzein (7,4′-dihydroxyisoflavonone, 5), (d) genistein (5,7,4′-trihydroxyisoflavone, 8), (e) naringenin (5,7,4′-trihydroxyflavonone, 9), (f) olivetol (12), and (g) resveratrol (3,4′,5-trihydroxystilbene, 13). These prenyl acceptors gave the following reaction products:
(a) 5-geranyl-1,6-DHN and 2-geranyl-1,6-DHN;
(b) 1-geranyl-2,7-DHN and 1,6-digeranyl-2,7-DHN;
(c) 7-O-geranyl-daidzein;
(d) 7-O-geranyl-genistein;
(e) 6-geranyl-naringenin and 7-O-geranyl-naringenin;
(f) 2-geranyl-olivetol and 4-geranyl-olivetol, and
(g) 4-geranyl-resveratrol.
Example 6
Mg2+ Dependent Prenyltransferase Activity of ORF2
The basal reaction buffer employed contained 50 mM HEPES (pH 7.5), 5 mM DTT (as needed), 5 mM prenyl acceptor, DHN2, and 5 mM GPP, in a final volume of 20 μl. The magnesium-containing reaction mixture contained 5 mM MgCl2. The reaction was initiated by adding 20 μg of ORF2 protein to the basal assay mixture. After incubation at room temperature for 4 hrs., the reaction mixture was dried using SpeedVac and the dried residue was spotted on a silica gel TLC plate. The plate was developed with chloroform:methanol (15-30:1). Reaction products were detected at 254 nm UV.
In the presence of magnesium in the reaction mixture, prenylated products were readily observed, while in the absence of magnesium in the reaction mixture, no prenylated products were observed.
Example 7
Promiscuous Activity of ORF2 with Different Flavonoids and Other Compounds
The reaction buffer employed contained 50 mM HEPES (pH 7.5), 5 mM DTT (as needed), 5 mM MgCl2, 0.1 mM of each prenyl acceptor, 0.1 mM GPP, and 0.01 mM [14C]GPP in a final volume of 20 μl. The reaction was initiated by adding 20 μg of ORF2 protein to the assay mixture. After incubation at room temperature for 4 hrs., the reaction mixture was dried using SpeedVac and the dried residue was spotted on a silica gel TLC plate. The plate was developed with chloroform:methanol (15-30:1). Reaction products were detected with a phosphoimager. The compounds tested were daidzein, fisetin, formononetin, genistein, naringenin, 4-HPP and DHN2.
With each of the prenyl acceptors tested, prenylated products were readily observed.
Example 8
Knock Out ORF2 Mutant in Streptomyces sp. Strain CL190
To gain insight into the function of ORF2, an ORF2 knock out mutant was constructed by frame-shift mutation into orf2. Thus, the 9.0 kb-BamHI-BamHI DNA fragment containing orf2 was cloned into BamHI site of pUC118 (Takara, Kyoto, Japan), a vector of E. coli. The resulting plasmid, pCL3301, was digested with EcoRI and then a 3.5-kb EcoRI-EcoRI DNA fragment was cloned into EcoRI site of pUC118 to give pCL3301E3. In addition, a 2.0-kb BamHI-EcoRI DNA fragment in pCL3301 was cloned into BamHI-EcoRI site of pBluescript (Toyobo, Tsuruga, Japan) to give pCL3301BE2. pCL3301E3 was digested with BglII, the recognition site of which is in the targeted orf2, and then blunt-ended with T4 DNA polymerase (Takara). Next, this blunt-ended DNA fragment was self-ligated to give pCL3301E3Bg, which contains orf2 having a frame shift mutation. A 3.5-kb EcoRI-EcoRI DNA fragment cut from pCL3301E3Bg was cloned into the EcoRI site of pCL3301BE2 to give pBluedORF2. Finally, a 5.0-kb XbaI-KpnI fragment, both the recognition sites of which are in the vector pBluescript, was ligated into the same sites of pSE101 (see Dairi et al., Biosci Biotechnol Biochem 59:1835-1841 (1995)), a Streptomyces-E. coli shuttle vector, to give pSEdORF2.
Streptomyces sp. strain CL190 was transformed with pSEdORF2 (as described by Kieser, et al., in “Practical Streptomyces Genetics”, eds. The John Innes Foundation, Norwich (2000). General considerations about gene cloning in Streptomyces. pp. 211-228) and a desired transformant was selected on R2YE plates containing 20 μg thiostrepton/ml. Next, the transformant was cultivated in SK2 liquid medium containing 20 μg thiostrepton/ml at 30° C. for 3 days. As described by Kieser et al., supra, protoplast was prepared from the transformant mycelium and regenerated on R2YE medium without thiostrepton. Each regenerated colony was simultaneously inoculated on Bennet plates with and without thiostrepton and a thiostrepton sensitive colony was selected to obtain the ORF2 knocking out mutant, Streptomyces sp. strain CL190 dORF2-8. It was confirmed by PCR that the mutant actually had frame-shift mutation in orf2 (FIG. 7).
The constructed mutant and CL190 were cultivated as reported by Shin-ya, et al., in J. Antibiot. (Tokyo) 43, 444-447 (1990). Myceria were harvested by centrifuge and then naphterpin produced by CL190 was extracted from the CL190 mycerium by the same method previously reported (Shin-ya et al., supra). The mycerium of the mutant was also extracted by the same method. Both the extracts were analyzed on silica gel-thin layer chromatography (TLC) as described. As a result, naphterpin was detected in the extract from CL190, but not in the extract from the mutant (FIG. 7). This result unequivocally indicates that ORF2 is essential for the naphterpin biosynthesis.
Example 9
Database Searches
Data base searches for sequence and structural homologues were performed with PSI-BLAST and VAST (accessible via the internet on the world wide web at the URL “ncbi.nlm.nih.gov”), SSM and DALI (accessible via the internet on the world wide web at the URL “ebi.ac.uk/msd-srv/ssm”), CE (accessible via the internet on the world wide web at the URL “cl.sdsc.edu/ce.html”) and DEJAVU (accessible via the internet on the world wide web at the URL “portray.bmc.uu.se/”) respectively, through the Protein Data Bank (accessible via the internet on the world wide web at the URL “rcsb.org/pdb/”), the Structural Classification of Proteins (SCOP, accessible via the internet on the world wide web at the URL “scop.mrc-lmb.cam.ac.uk/scop”), and the CATH Protein structure classification (accessible via the internet on the world wide web at the URL “biochem.ucl.ac.uk/bsm/cath”).
Example 10
Modeling
Models of CloQ/NovQ and HypSc were performed with the Modeller-4 package (Sali et al., Proteins 23(3):318-26 (1995)) using Orf2 as a structural template (see FIG. 6). For each sequence, five different models were calculated and evaluated. The multiple sequence alignment was then hand modified based on the superposition of the Orf2 structure with the different models. Modeller was then re-run and the iteratively generated models visually inspected and adjusted if necessary. Model quality was assessed with PROCHECK (see Laskowski, et al., J Appl Cryst 26:283-291 (1993)). Side chains presenting potentially significant variation between the differenct active sites are displayed and labeled. Conserved residues in the different models include Asp 110, Lys 119, Asn 173, Tyr 175, Tyr 216, and Arg 228, of which only Asp 110 and Arg 228 are displayed for clarity.
Example 11
Comparative Modeling of CloQ/NovQ and hypSc
While a significant degree of active site residue correspondence is consistent with the validity of the homology models, small but critical differences in key active site residues provide reasons for the shorter prenyl chain length specificity of CloQ/NovQ and for differences in aromatic substrate selectivity (Pojer et al., supra). In addition, the homology model of HypSc is consistent with the hypothesis for prenyl chain length specificity in this predicted protein. Tyr 121, involved in GPP binding in Orf2, is replaced in all other sequences by a Trp (115 in CloQ/NovQ, and 117 in HypSc): the modeled ring orientation is identical to Tyr 121 while the increased bulkiness may better sequester the shorter C5 prenyl chain of DMAPP. Ser 64 and Gly 286, replaced by Arg (59 in CloQ/NovQ, 61 in HypSc) and Glu (274 in CloQ/NovQ/HypSC), respectively, appear poised to form an internal salt-bridge precisely over the location of the second C5 isoprene unit of the GPP molecule experimentally positioned in the Orf2 active site. Identical changes observed in the HypSc prenyltransferase model predict that this enzyme will also use DMAPP as a prenyl donor. Notably, in Orf2, the geranyl chain of the GPP molecule ends next to the barrel opening, thus providing a probable reason for Orf2's ability to accommodate the C15 prenyl chain of the FPP unit.
Moreover, structural alignment of Orf2 with the CloQ/NovQ and HypSc models reveals the molecular determinants for Orf2's requirement for divalent cations. Asp 62, directly involved in Orf2's diphosphate binding via a coordinated Mg2+ ion, is conservatively replaced in HypSc by Asn 59 but changes to a Ser 57 in CloQ/NovQ. In a complementary manner, Ser 51 in Orf2 is replaced by a positively charged Lys 47 in CloQ/NovQ and a positively charged Arg 47 in HypSc that, with little rotamer rearrangement, can be positioned over the Mg2+ ion observed in Orf2. Furthermore, these basic side chains are ideally positioned for electrostatic binding to the negatively charged α-phosphate of the GPP molecule. However, Asp 110, involved indirectly in binding Mg2+ via a water molecule, is conserved in all the sequences examined (FIG. 1B); thus providing an explanation as to why CloQ and NovQ are active in the absence of Mg2+ but display maximum activity in the presence of 2.5 mM Mg2+ (Pojer et al, supra). Regarding CloQ's specificity for 4-HPP, Orf2's Gln 161 and Ser 177 are replaced by Arg, 153 and 169 in CloQ, and are positioned to possibly bind the negatively charged tail of the 4-HPP substrate.
This analysis allows one to predict that the HypSc enzyme is a prenyltransferase, accepting only DMAPP as a substrate, while not requiring Mg2+ for its activity; this homology modeling based hypothesis has been confirmed by the cloning, protein expression and enzymatic assays of HypSc (see FIG. 7).







TABLE 3







Crystallographic data, phasing, and refinement statistics










SeMet-Orf2

















λ1 (inf,
λ 2 (peak,
λ 3
Wt +

Wt + GSPP +
wt + GSPP +



Data Set
max f′′)
min f′)
(remote)
TAPS
Wt + GPP
1,6-DHN
Flaviolin


















Beam line
BNL-X8C
BNL-X8C
BNL-X8C
BNL-X6A
Salk Inst.
ESRF-BM30A
SSRL-9.1



Wavelength (Å)
0.9793
0.97915
0.9641
0.934
1.54178
0.9797


Space Group
P21212
P21212
P21212
P21212
P21212
P21212
P21212


Unit cell a, b, c (Å)
71.3, 91.2,
71.4, 91.2,
71.3, 91.1,
71.3, 91.2,
74.6, 91.9,
71.3, 90.2,
73.6, 91.6,



48.3
48.4
48.3
48.3
48.8
47.5
48.6


Resolution (Å)
  50-1.55
  50-1.55
  50-1.50
  50-1.45
  99-2.25
  99-1.95
  50-2.02


last shell (Å)
1.61-1.55
1.57-1.52
1.55-1.50
1.42-1.40
2.29-2.25
2.00-1.95
2.09-2.02


Observations


Overalla
156510
162052
170494
139582
70572
115225
110252


Uniquea
86828
91336
95274
49390
16155
22960
21790


Redundancya,b
1.8 (1.9)
1.8 (1.5)
1.8 (1.5)
2.84 (1.94)
4.4 (4.2)
5.0 (5.1)
4.5 (5.0)


Completenessa,b(%)
98.2 (98.2)
97.5 (91.1)
  98 (94.8)
78.3 (50.4)
  98 (99.9)
99.5 (99.6)
99.1 (99.5)


I/σ Ib
15.3 (2.2) 
14.7 (1.7) 
14.3 (1.7) 
37.34 (38.9) 
13.5 (2.5) 
32.3 (35.5)
31.9 (38.0)


Rsymb,c(%)
 7.4 (49.9)
 6.6 (48.3)
 7.2 (55.7)
 9.7 (51.5)
 8.6 (54.8)
 8.6 (51.2)
 9.0 (57.2)


No. of Se sites

7


FOMd


centric
0.57


acentric
0.55


Rcryste/Rfreef(%)

21.42 (24.22)


23.0 (25.8)
22.25 (25.1) 
24.1 (27.1)
23.0 (26.8)


Missing residues

5


6
5
6
6


Protein atoms

2322


2332
2338
2332
2332


Water molecules

254


429
205
346
320


Ions boundg

0


0
3
2
3


Substrate and/or




15
19
31
32


binding agent


atomsh
0


R.m.s.d. bond

0.005


0.005
0.006
0.006
0.007


length (Å)


R.m.s.d. bond

1.2


1.2
1.2
1.2
1.1


angles (°)


average B-


factor(Å2)


protein

13.6


16.2
28.2
43.5
29.4


water

22.8


27.7
35.3
42.5
42.6


substrate and/or

0


20.5
67.4
52.5
41.0


binding agent





aFor the SeMet data sets, F+and F− were considered non-equivalent when calculating the number of unique reflections and completeness.


bNumber in parenthesis is for highest resolution shell.


cRsym = Σh|Ih − <Ih>|/Σh(Ih), where <Ih> is the average intensity over symmetry equivalent reflections.


dFOM is the Figure of Merit


eRcryst = Σ| | Fobs − Fcalc| |/Σ|Fobs|, where summation is over the data used for refinement.


fRfree factor is Rcryst calculated using 5% of data (test set) excluded from refinement.


gIon bounds refers to Mg2+, (NO3)2− ions.


hSubstrate and/or binding agent atoms refers to TAPS, GPP, GSPP and 1,6-DHN, GSPP and flaviolin molecules.






Thus, Table 3 summarizes the structural features accompanying prenyl chain length determination, aromatic substrate selectivity and the mechanism of prenyl group transfer, as determined by obtaining X-ray crystal structures of four Orf2 substrate/substrate analogue complexes, namely Orf2 complexed with a TAPS buffer molecule, a binary Orf2 complex containing GPP and Mg2+, a ternary Orf2 complex with a non-hydrolyzable GPP analogue, GSPP, Mg2+ and 1,6-DHN, and a ternary Orf2 complex with GSPP, Mg2+ and flaviolin.
Example 12
Detection of Prenyltransferase Activity of hypSc
The assay described in Example 5 was repeated with hypSc and (a) 1,6-DHN (2), (b) 2,7-DHN (3), (c) daidzein (7,4′-dihydroxyisoflavonone, 5), (d) genistein (5,7,4′-trihydroxyisoflavone, 8), (e) naringenin (5,7,4′-trihydroxyflavonone, 9), (f) olivetol (12), and (g) resveratrol (3,4′,5-trihydroxystilbene, 13). These prenyl acceptors gave the following reaction products:
(a) 5-dimethylallyl-1,6-DHN;
(b) 1-dimethylallyl-2,7-DHN;
(c) no reaction products detected;
(d) no reaction products detected;
(e) 6-dimethylallyl-naringenin;
(f) 2-dimethylallyl-olivetol and 4-dimethylallyl-olivetol, and
(g) 4-dimethylallyl-resveratrol.
Example 13
Biosynthesis of Hybrid Isoprenoids from Marine Actinomycetes
With the PTases Orf2 and HypSc in hand, additional actinomycete PTases with different substrate specificities can be identified. To this end, a group of marine actinomycetes that produce assorted hybrid isoprenoid natural products was compiled (see FIG. 1B, Table 4).







TABLE 4







Hybrid isoprenoid-producing actinomycetes















arom.


Strain
natural product
attachment
isoprene
substrate





S. sp. CL190
naphterpin
C
GPP
hydroxy-






naphthalene


CNB632
marinone +
C
FPP
hydroxy-



analogs


naphthalene


CNH099
marinone
C
FPP
hydroxy-






naphthalene



neomarinone
C
FPP
hydroxy-






naphthalene



lavanducyanin
N
GPP
phenazine


CNQ525
Q525.518
C × 2
DMAPP/
hydroxy-





GPP
naphthalene


CNQ509
Q509.364
O
GPP
phenazine



Q509.366
C
FPP
nitropyrrole


S. purpeofuscus
aestivophoenins
N and C
DMAPP
phenazine









Strain CNH099 produces three isoprenoid chemotypes, namely the farnesylated naphterpin analog marinone, the rearranged derivative neomarinone and the phenazine lavanducyanin. Feeding experiments with labeled precursors delineated the biosynthetic course for these metabolites. The biosynthesis of the naphthoquinone core common amongst the marinones must proceed through a symmetrical pentaketide intermediate such as THN to satisfy the observed labeling patterns. Flaviolin, a known auto-oxidation product of THN, either directly or methylated at C10 via S-adenosyl methionine may serve as an intermediate in neomarinone biosynthesis. FPP, derived from the MEP pathway, provides the sesquiterpenoid side chain. Prenylation may occur directly via C-prenylation though attachment of C3 of FPP or indirectly via O-prenylation of the C5 or C7-hydroxy groups of flaviolin followed by Claisen rearrangement to yield the same furan intermediate. Proton assisted cyclization of the linear diene following Wagner-Meerwein rearrangements yields neomarinone.
A preliminary search for the respective biosynthetic gene clusters allows the generation of a cosmid library in the E. coli-Streptomyces shuttle cosmid pOJ446, the development of a genetics system in this strain for homologous recombination involving the E. coli to CNH099 conjugal transfer of pKC1139-based temperature-sensitive plasmids, and the sequence analysis of genes encoding THN and phenazine biosynthesis. Additionally a pOJ446 cosmid library of the aestivophoenin producer Streptomyces purpeofuscus has been prepared. This information is useful for the identification and cloning of novel aromatic PTases.
Example 14
X-Ray Crystallographic Structures of Orf2 Complexed to Geranylated Products
The reaction products of Orf2 incubated with GPP and 1,6-DHN and naringenin, respectively, have been identified as trans-5-geranyl 1,6-DHN/trans-2-geranyl 1,6-DHN and 6-geranyl naringenin/7-O-geranyl naringenin, respectively (see FIG. 2B). Large scale production of these compounds can be carried out in vitro using 500 uL reaction volumes in the assay buffer described herein and incorporating 20-50 mM GPP and 20-50 mM 1,6-DHN or (2S)-naringenin. Incubations can be carried out overnight and a sample of the resultant solution analyzed by HPLC-MS to assess the product yield and extent of reaction. Multiple reactions can be combined (approximately 5-10 individual reactions), extracted two times with equal volumes of ethyl acetate each time, the combined organic extracts dried down, then dissolved in a minimal amount of methanol followed by injection on an HPLC and purified on a preparative reverse phase column. Purified products can then be characterized. Purified trans-5-geranyl 1,6-DHN, trans-2-geranyl 1,6-DHN, 6-geranyl naringenin, and 7-O-geranyl naringenin can be dissolved in 100% ethanol or methanol to near saturation (approximately 100-200 mM). Each of the four Orf2 product complexes can be prepared employing co-crystallization and soaking strategies. To ensure the maximal occupancy of the product in Orf2 crystals, both co-crystallization and soaking approaches employ a grid whereby the concentrations of the respective products is varied between 5 and 25 mM.
Example 15
Creating an Orf2 Mutant Capable of Efficient Use of DMAPP and Elucidate Its Three Dimensional Structure in the Presence of DMASPP and 1,6-DHN
In order to further define prenyl diphosphate chain length selectivity, molecular determinants of aromatic substrate recognition and divalent cation dependence, homology modeling of CloQ, NovQ and HypSc sequences were carried out using the three dimensional coordinates of Orf2 as a structural template (FIG. 6). While the degree of active site residue correspondence is consistent with the homology models discussed above, small differences in key active site residues may explain the shorter prenyl chain length specificity of CloQ/NovQ and the differences in aromatic substrate selectivity. In addition, the homology model of a newly identified PTase from Streptomyces coelicolor, HypSc, lead to the biochemical characterization of HypSc as a DMAPP-specific, Mg2+-independent PTase. Prenyl chain length dependence in Orf2 can be evaluated in a variety of ways. One approach involves the generation of a limited set of site directed mutants based upon the initial homology models of CloQ/NovQ and HypSc shown in FIG. 6. An alternate approach involves the generation of several 1024 member mutant libraries of all possible amino acid permutations derived from the comparative analysis of Orf2 with either HypSc or CloQ/NovQ and centered around the geranyl binding site mapped previously.
The first set of Orf2 mutants can be constructed using a traditional QuickChange protocol. Specifically, a Trp residue (residue 115 in CloQ/NovQ and residue 117 in HypSc) replaces Tyr 121 in Orf2. The increased bulkiness of the indole ring in HypSc/CloQ/NovQ compared to the phenolic ring in Orf2 may better accommodate the shorter C5 prenyl chain of DMAPP. In addition, in HypSc/CloQ/NovQ, Arg and Glu residues replace Ser 64 and Gly 286, respectively in Orf2 (residues 59 and 274 in CloQ/NovQ and residues 61 and 274 in HypSc). This apparent salt bridge in the DMAPP-specific PTases sits poised over the location of the second C5 isoprene unit of the GPP molecule. Notably, this change in the homology model of HypSc lead to the biochemical characterization of this newly discovered S. coelicolor enzyme as a DMAPP specific PTase. This rather directed approach towards enzyme engineering minimizes the potential influence of neighboring residues towards prenyl chain length determination. If this mutagenic strategy fails to significantly alter Orf2's prenyl chain length specificity, a larger library of mutant Orf2s can be prepared employing the SCOPE approach described in Example 17.
Comparative homology modeling used to initially select residues for further functional examination is performed with the package Modeller-4 using Orf2 as a structural template. As new sequences are identified, additional models can be constructed. For each sequence, five different models are calculated and evaluated. The multiple sequence alignment is then modified by hand based on the superposition of the Orf2 structure with the individual models. Modeller-4 is then re-run and the iteratively generated models visually inspected and adjusted again if necessary. Model quality is assessed with PROCHECK.
Example 16
Development of a Quantitative PTase Kinetic Assay
To determine the steady state kinetic parameters for PTases, a radiometric TLC assay can be employed. The typical reaction buffer contemplated for use consists of 50 mM HEPES (pH 7.5), 0.1-10 mM aromatic acceptor, 0.1-5 mM [14C]-DMAPP, [14C]-GPP or [14C]-FPP (New England Nuclear), 5 mM MgCl2 in a final volume of 20 μl. The reaction is initiated by adding 10 ng-5 μg of PTase to the assay mixture. Enzyme concentration ranges can be selected to achieve the optimal PTase concentration obeying Michaelis-Menten kinetics. Incubations can be carried out and 4-6 time points collected in triplicate over an initial time range of 1-120 minutes. Reactions can be quenched with ethyl acetate. Extracts can be evaporated to dryness, re-dissolved in methanol, and applied to Whatman LK6D silica TLC plates. The TLC plate can be developed with a chloroform/methanol (20:1) solvent mixture. Aromatic reaction products can be detected at 254 nm or by autoradiography using imaging plates. Products can be quantified by scraping sections of the TLC plate into Ecolume scintillation fluid, detecting [14C]-radioactivity with a scintillation counter, and converting the corrected cpm into nmoles of product using the final specific radioactivity of [14C]-DMAPP, [14C]-GPP or [14C]-FPP. Kinetic constants can be determined from initial velocity measurements, in which product formation is linear over the time periods monitored (up to two hours for low activity PTases or mutants thereof). Given the fact that two substrates are employed, KM values for the prenyl donor are established using saturating concentrations of aromatic acceptor (typically 50 mM) and KM values for the aromatic acceptors are established using saturating concentrations of prenyl donors, typically 50 mM. In order to reach 50 mM prenyl donor, the radioactive sample is diluted to 50 mM using cold DMAPP, GPP or FPP and corrections for dilution applied as appropriate.
Example 17
A Rapid UPLC-MS-Based Qualitative Assay to Monitor Prenyl Group Transfer
An efficient analysis technique is desirable to serve as a qualitative (or semi-quantitative) assay for prenylation reactions. Specifically, biosynthetic transformations to be monitored include the prenylation (via IPP, GPP and FPP) of both natural and unnatural substrates for wild-type as well as mutant PTase. Efficiency for such an assay is defined in terms of speed, resolution and sensitivity. The assay must accommodate large numbers of samples (high-throughput) for evaluating the several 1024 mutant libraries and must provide analysis on low volume (sub milliliter) reaction volumes (given the number of reactions and the associated costs for reagents). In addition to screening these enzymes and enzyme libraries against natural substrates with a selection of isoprenoid diphosphates, screening may also be desirable with respect to various unnatural substrates designed to probe the structure-to-reactivity relationships governing regio-specific prenylation of chemical building blocks. Finally, given the established promiscuity of Orf2 and its ability to generate multiple products, the assay must also resolve and identify multiple prenylated species per reaction.
Given these requirements, Ultra Performance Liquid Chromatography coupled with ESCi Mass Spectroscopy (UPLC-MS) has been identified as a suitable technique to satisfy the above described assay needs. Briefly, UPLC is a recent advancement in separations technology. The new 1.7 μm particle technology coupled with operating pressures approaching 15,000 psi results in gains of 1.7× in resolution, 3× in speed and 1.7× in sensitivity versus standard HPLC using 5 μm particles when column lengths are normalized. However, the greatest benefit of this technology is realized when normalizing resolution (L/dp); here the gains are 1× in resolution, 9× in speed and 3× in sensitivity versus traditional HPLC 5 μm particles. These significant gains in speed and sensitivity are very beneficial for achieving a qualitative assay.
An additional benefit of this platform stems from the 500% reduction in time for methods development. Preferably, the experiments are carried out using micro-well plates (96 or 384-well format) where the PTases, isoprenoid diphosphate, and aromatic substrate are sequentially added from stock solutions and mixed. Libraries of mutant enzymes are conveniently purified in parallel using small scale (5-10 ml) cultures and an automated Qiagen robot for the parallel purification of histidine-tagged proteins. Following the optimal reaction time, the reactions are quenched and loaded into the UPLC sample organizer/manager for assay. The target operating parameters include LC run times <5 min. (run times as short as 1 min. may be attainable) and direct injection of the quenched reaction mixtures to eliminate sample loss issues. If direct injection is not feasible for high-throughput, an extraction step using a less polar organic solvent can be included, and then sample can be taken directly from the top layer of the reaction well, even while covered to address any evaporation and subsequent sample concentration issues.
Finally, product detection can be achieved via diode array UV detection triggering MS acquisition. Because injection volumes as low as 0.1 μl are possible, and retention times are so short, total volumes of <5 ml/sample are run directly through the ESCi MS assuring all peaks are detected. With the parent ion information, the identification of prenylated products will be facile. The Mass Lynx data management system permits the automation of data analysis, quickly identifying peaks of interest by predefining product mass tables.
Example 18
Structure Elucidation
Large scale in vitro reactions and whole cell fermentations can be directly analyzed on a Waters 600 HPLC or a Agilent 1100 HPLC equipped with photodiode array detection (PDA), auto-sampling and fraction collection. Separations are achieved using a YMC ODS-AQ 4.6×150 mm reversed-phase column with a linear solvent gradient of 0.15% TFA in water to methanol over 30 min at a flow rate of 0.5 ml/min. Alternatively, the samples are first extracted with ethyl acetate, dried over MgSO4, filtered, dried, and redissolved in methanol for analysis. When possible, chromatographic peaks are identified by co-injection with authentic standards. Automated screening will be carried out on a Waters Acquity UPLC equipped with PDA detection and an in-line MicroMass ZQ ESCi (combination APCI-ESI) Mass Spectrometer for low resolution mass analysis. Isolation of pure constituents are carried out with pre-fractionated samples a 20×250 mm YMC pack ODS-A HPLC column that can operate at a flow rate of up to 10 ml/min.
Structures of pure metabolites can be elucidated by 1D and 2D-NMR spectroscopy on Bruker DRX-300 and DRX-600 spectrometers, or on a Varian Unity Inova 500 Spectrometer. Proton and carbon assignments can be obtained from COSY, HSQC, HMBC, and nOe spectral data. Homonuclear 1H connectivities can be determined by the phase-sensitive, double-quantum filtered COSY experiment. One-bond heteronuclear 1H-13C connectivities can be determined by gradient-enhanced proton-detected HSQC experiments. Two- and three-bond 1H-13C connectivities can be determined by gradient-enhanced proton-detected HMBC experiments. Homonuclear 1H nOe's can be obtained by difference nOe experiments and by two-dimensional ROESY experiments to generate relative stereochemistry while the absolute stereochemistry of new compounds can often be achieved through the modified Mosher analytical method or single crystal X-ray analysis. When appropriate, biosynthetic intermediates labeled with stable isotopes (such as sodium [1,2-13C2]acetate or [U-13C6]glucose) can be administered to the cultures to aid in analog identification through the 13C INADEQUATE or related experiment. High-resolution mass determination can be performed by TOF-ESI (TSRI Mass Spectroscopy Laboratory) or FAB. Additional characterization techniques include Polarimetry (Perkin-Elmer 341 Polarimeter) and Fourier-Transform Infrared Spectroscopy (Nicolet 4700 FT-IR).
While the invention has been described in detail with reference to certain preferred embodiments thereof, it will be understood that modifications and variations are within the spirit and scope of that which is described and claimed.







APPENDIX 1





Atomic coordinates of aromatic prenyltransferase residues 3-302 of SEQ ID NO: 2.
















PROGRAM:
CNS 1.1


AUTHORS:
BRUNGER, ADAMS, CLORE, DELANO, GROS, GROSSE-KUNSTLEVE, JIANG, KUSZEWSKI,



NILGES, PANNU, READ, RICE, SIMONSON, WARREN







DATA USED IN REFINEMENT.










RESOLUTION RANGE HIGH (ANGSTROMS):
1.95



RESOLUTION RANGE LOW (ANGSTROMS):
29.74



DATA CUTOFF (SIGMA(F)):
0.0



DATA CUTOFF HIGH (ABS(F)):
1386159.12



DATA CUTOFF LOW (ABS(F)):
0.000000



COMPLETENESS (WORKING + TEST) (%):
99.4



NUMBER OF REFLECTIONS:
22923







FIT TO DATA USED IN REFINEMENT.










CROSS-VALIDATION METHOD:
THROUGHOUT



FREE R VALUE TEST SET SELECTION:
RANDOM



R VALUE (WORKING SET):
0.241



FREE R VALUE:
0.271



FREE R VALUE TEST SET SIZE (%):
5.0



FREE R VALUE TEST SET COUNT:
1154



ESTIMATED ERROR OF FREE R VALUE:
0.008







FIT IN THE HIGHEST RESOLUTION BIN.










TOTAL NUMBER OF BINS USED:
6



BIN RESOLUTION RANGE HIGH (A):
1.95



BIN RESOLUTION RANGE LOW (A):
2.07



BIN COMPLETENESS (WORKING + TEST) (%):
99.7



REFLECTIONS IN BIN (WORKING SET):
3569



BIN R VALUE (WORKING SET):
0.352



BIN FREE R VALUE:
0.386



BIN FREE R VALUE TEST SET SIZE (%):
4.9



BIN FREE R VALUE TEST SET COUNT:
184



ESTIMATED ERROR OF BIN FREE R VALUE:
0.028







NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.










PROTEIN ATOMS:
0



NUCLEIC ACID ATOMS:
0



HETEROGEN ATOMS:
0



SOLVENT ATOMS:
0







B VALUES.










FROM WILSON PLOT (A**2):
25.5



MEAN B VALUE (OVERALL, A**2):
43.5



OVERALL ANISOTROPIC B VALUE.



B11 (A**2):
−3.35



B22 (A**2):
−13.96



B33 (A**2):
17.30



B12 (A**2):
0.00



B13 (A**2):
0.00



B23 (A**2):
0.00







BULK SOLVENT MODELING.










METHOD USED:
FLAT MODEL



KSOL:
0.355809



BSOL:
42.5443 (A**2)







ESTIMATED COORDINATE ERROR.










ESD FROM LUZZATI PLOT (A):
0.30



ESD FROM SIGMAA (A):
0.35



LOW RESOLUTION CUTOFF (A):
5.00







CROSS-VALIDATED ESTIMATED COORDINATE ERROR.










ESD FROM C-V LUZZATI PLOT (A):
0.35



ESD FROM C-V SIGMAA (A):
0.36







RMS DEVIATIONS FROM IDEAL VALUES.










BOND LENGTHS (A):
0.006



BOND ANGLES (DEGREES):
1.2



DIHEDRAL ANGLES (DEGREES):
24.4



IMPROPER ANGLES (DEGREES):
0.89








ISOTROPIC THERMAL MODEL:
RESTRAINED


ISOTROPIC THERMAL FACTOR RESTRAINTS.
RMS SIGMA










MAIN-CHAIN BOND (A**2):
NULL; NULL



MAIN-CHAIN ANGLE (A**2):
NULL; NULL



SIDE-CHAIN BOND (A**2):
NULL; NULL



SIDE-CHAIN ANGLE (A**2):
NULL; NULL








NCS MODEL:
NONE


NCS RESTRAINTS.
RMS SIGMA/WEIGHT










GROUP 1 POSITIONAL (A):
NULL; NULL



GROUP 1 B-FACTOR (A*2):
NULL; NULL








PARAMETER FILE 1:
CNS_TOPPAR/protein_rep.param


PARAMETER FILE 2:
CNS_TOPPAR/dna-rna_rep.param


PARAMETER FILE 3:
CNS_TOPPAR/water_rep.param


PARAMETER FILE 4:
CNS_TOPPAR/ion.param


PARAMETER FILE 5:
CNSPAR:gspp_dhn2_no3.param


TOPOLOGY FILE 1:
CNS_TOPPAR/protein.top


TOPOLOGY FILE 2:
CNS_TOPPAR/dna-rna.top


TOPOLOGY FILE 3:
CNS_TOPPAR/water.top


TOPOLOGY FILE 4:
CNS_TOPPAR/ion.top


TOPOLOGY FILE 5:
CNSPAR:gspp_dhn2_no3.top







OTHER REFINEMENT REMARKS: NULL























SEQRES
1
A
514
GLU
ALA
ALA
ASP
VAL
GLU
ARG
VAL
TYR
ALA
ALA
MET
GLU


SEQRES
2
A
514
GLU
ALA
ALA
GLY
LEU
LEU
GLY
VAL
ALA
CYS
ALA
ARG
ASP


SEQRES
3
A
514
LYS
ILE
TYR
PRO
LEU
LEU
SER
THR
PHE
GLN
ASP
THR
LEU


SEQRES
4
A
514
VAL
GLU
GLY
GLY
SER
VAL
VAL
VAL
PHE
SER
MET
ALA
SER


SEQRES
5
A
514
GLY
ARG
HIS
SER
THR
GLU
LEU
ASP
PHE
SER
ILE
SER
VAL


SEQRES
6
A
514
PRO
THR
SER
HIS
GLY
ASP
PRO
TYR
ALA
THR
VAL
VAL
GLU


SEQRES
7
A
514
LYS
GLY
LEU
PHE
PRO
ALA
THR
GLY
HIS
PRO
VAL
ASP
ASP


SEQRES
8
A
514
LEU
LEU
ALA
ASP
THR
GLN
LYS
HIS
LEU
PRO
VAL
SER
MET


SEQRES
9
A
514
PHE
ALA
ILE
ASP
GLY
GLU
VAL
THR
GLY
GLY
PHE
LYS
LYS


SEQRES
10
A
514
THR
TYR
ALA
PHE
PHE
PRO
THR
ASP
ASN
MET
PRO
GLY
VAL


SEQRES
11
A
514
ALA
GLU
LEU
SER
ALA
ILE
PRO
SER
MET
PRO
PRO
ALA
VAL


SEQRES
12
A
514
ALA
GLU
ASN
ALA
GLU
LEU
PHE
ALA
ARG
TYR
GLY
LEU
ASP


SEQRES
13
A
514
LYS
VAL
GLN
MET
THR
SER
MET
ASP
TYR
LYS
LYS
ARG
GLN


SEQRES
14
A
514
VAL
ASN
LEU
TYR
PHE
SER
GLU
LEU
SER
ALA
GLN
THR
LEU


SEQRES
15
A
514
GLU
ALA
GLU
SER
VAL
LEU
ALA
LEU
VAL
ARG
GLU
LEU
GLY


SEQRES
16
A
514
LEU
HIS
VAL
PRO
ASN
GLU
LEU
GLY
LEU
LYS
PHE
CYS
LYS


SEQRES
17
A
514
ARG
SER
PHE
SER
VAL
TYR
PRO
THR
LEU
ASN
TRP
GLU
THR


SEQRES
18
A
514
GLY
LYS
ILE
ASP
ARG
LEU
CYS
PHE
ALA
VAL
ILE
SER
ASN


SEQRES
19
A
514
ASP
PRO
THR
LEU
VAL
PRO
SER
SER
ASP
GLU
GLY
ASP
ILE


SEQRES
20
A
514
GLU
LYS
PHE
HIS
ASN
TYR
ALA
THR
LYS
ALA
PRO
TYR
ALA


SEQRES
21
A
514
TYR
VAL
GLY
GLU
LYS
ARG
THR
LEU
VAL
TYR
GLY
LEU
THR


SEQRES
22
A
514
LEU
SER
PRO
LYS
GLU
GLU
TYR
TYR
LYS
LEU
GLY
ALA
TYR


SEQRES
23
A
514
TYR
HIS
ILE
THR
ASP
VAL
GLN
ARG
GLY
LEU
LEU
LYS
ALA


SEQRES
24
A
514
PHE
ASP
MG2
GSP
DH2
NO3
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
25
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
26
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
27
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
28
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
29
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
30
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
31
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
32
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
33
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
34
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
35
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
36
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
37
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
38
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
39
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP
TIP


SEQRES
40
A
514
TIP
TIP
TIP
TIP
TIP
TIP
TIP







CRYST1   71.319  90.243  47.513  90.00  90.00  90.00  P 21 21 2  4












ORIGX1
1.000000
0.000000
0.000000
0.00000



ORIGX2
0.000000
1.000000
0.000000
0.00000


ORIGX3
0.000000
0.000000
1.000000
0.00000


SCALE1
0.014022
0.000000
0.000000
0.00000


SCALE2
0.000000
0.011081
0.000000
0.00000


SCALE3
0.000000
0.000000
0.021047
0.00000


















ATOM
1
CB
GLU
A
3
34.602
9.738
21.871
1.00
67.30
A


ATOM
2
CG
GLU
A
3
34.388
9.619
23.371
1.00
67.06
A


ATOM
3
CD
GLU
A
3
35.148
8.453
23.975
1.00
66.81
A


ATOM
4
OE1
GLU
A
3
36.396
8.490
23.969
1.00
66.45
A


ATOM
5
OE2
GLU
A
3
34.497
7.497
24.449
1.00
66.55
A


ATOM
6
C
GLU
A
3
32.569
8.406
21.275
1.00
68.45
A


ATOM
7
O
GLU
A
3
31.776
9.074
20.608
1.00
68.75
A


ATOM
8
N
GLU
A
3
34.407
8.713
19.628
1.00
67.62
A


ATOM
9
CA
GLU
A
3
34.078
8.542
21.070
1.00
67.93
A


ATOM
10
N
ALA
A
4
32.177
7.535
22.198
1.00
68.54
A


ATOM
11
CA
ALA
A
4
30.768
7.318
22.493
1.00
68.73
A


ATOM
12
CB
ALA
A
4
30.598
6.047
23.317
1.00
68.94
A


ATOM
13
C
ALA
A
4
30.212
8.517
23.257
1.00
69.02
A


ATOM
14
O
ALA
A
4
29.070
8.927
23.045
1.00
69.08
A


ATOM
15
N
ALA
A
5
31.036
9.076
24.139
1.00
68.87
A


ATOM
16
CA
ALA
A
5
30.649
10.222
24.953
1.00
67.96
A


ATOM
17
CB
ALA
A
5
31.752
10.542
25.956
1.00
67.47
A


ATOM
18
C
ALA
A
5
30.332
11.458
24.117
1.00
67.32
A


ATOM
19
O
ALA
A
5
29.330
12.131
24.356
1.00
67.46
A


ATOM
20
N
ASP
A
6
31.186
11.762
23.144
1.00
66.37
A


ATOM
21
CA
ASP
A
6
30.967
12.927
22.291
1.00
65.49
A


ATOM
22
CB
ASP
A
6
32.130
13.117
21.310
1.00
65.78
A


ATOM
23
CG
ASP
A
6
33.432
13.479
22.002
1.00
65.79
A


ATOM
24
OD1
ASP
A
6
33.412
14.365
22.887
1.00
65.52
A


ATOM
25
OD2
ASP
A
6
34.477
12.887
21.648
1.00
65.81
A


ATOM
26
C
ASP
A
6
29.667
12.802
21.506
1.00
64.46
A


ATOM
27
O
ASP
A
6
28.836
13.710
21.519
1.00
65.11
A


ATOM
28
N
VAL
A
7
29.493
11.675
20.824
1.00
63.25
A


ATOM
29
CA
VAL
A
7
28.292
11.442
20.031
1.00
61.69
A


ATOM
30
CB
VAL
A
7
28.331
10.054
19.351
1.00
60.95
A


ATOM
31
CG1
VAL
A
7
27.042
9.815
18.575
1.00
60.38
A


ATOM
32
CG2
VAL
A
7
29.530
9.968
18.423
1.00
60.39
A


ATOM
33
C
VAL
A
7
27.023
11.547
20.871
1.00
61.64
A


ATOM
34
O
VAL
A
7
26.055
12.185
20.458
1.00
61.60
A


ATOM
35
N
GLU
A
8
27.028
10.919
22.045
1.00
61.47
A


ATOM
36
CA
GLU
A
8
25.871
10.953
22.936
1.00
61.38
A


ATOM
37
CB
GLU
A
8
26.031
9.939
24.076
1.00
63.51
A


ATOM
38
CG
GLU
A
8
25.959
8.480
23.639
1.00
67.51
A


ATOM
39
CD
GLU
A
8
26.050
7.504
24.805
1.00
70.62
A


ATOM
40
OE1
GLU
A
8
25.181
7.560
25.706
1.00
71.88
A


ATOM
41
OE2
GLU
A
8
26.990
6.678
24.819
1.00
72.29
A


ATOM
42
C
GLU
A
8
25.662
12.345
23.523
1.00
59.87
A


ATOM
43
O
GLU
A
8
24.531
12.748
23.787
1.00
59.59
A


ATOM
44
N
ARG
A
9
26.755
13.072
23.729
1.00
58.71
A


ATOM
45
CA
ARG
A
9
26.681
14.418
24.285
1.00
57.97
A


ATOM
46
CB
ARG
A
9
28.077
14.907
24.679
1.00
57.94
A


ATOM
47
CG
ARG
A
9
28.108
16.329
25.214
1.00
58.66
A


ATOM
48
CD
ARG
A
9
29.481
16.687
25.761
1.00
59.63
A


ATOM
49
NE
ARG
A
9
30.534
16.601
24.752
1.00
60.75
A


ATOM
50
CZ
ARG
A
9
30.662
17.435
23.724
1.00
60.72
A


ATOM
51
NH1
ARG
A
9
29.798
18.430
23.561
1.00
59.60
A


ATOM
52
NH2
ARG
A
9
31.659
17.277
22.861
1.00
60.10
A


ATOM
53
C
ARG
A
9
26.054
15.381
23.281
1.00
57.24
A


ATOM
54
O
ARG
A
9
25.192
16.184
23.634
1.00
56.91
A


ATOM
55
N
VAL
A
10
26.488
15.298
22.029
1.00
56.17
A


ATOM
56
CA
VAL
A
10
25.948
16.163
20.989
1.00
54.31
A


ATOM
57
CB
VAL
A
10
26.769
16.052
19.688
1.00
54.27
A


ATOM
58
CG1
VAL
A
10
26.041
16.741
18.542
1.00
54.08
A


ATOM
59
CG2
VAL
A
10
28.130
16.691
19.891
1.00
53.91
A


ATOM
60
C
VAL
A
10
24.488
15.828
20.700
1.00
53.68
A


ATOM
61
O
VAL
A
10
23.661
16.725
20.541
1.00
53.84
A


ATOM
62
N
TYR
A
11
24.163
14.542
20.636
1.00
52.98
A


ATOM
63
CA
TYR
A
11
22.786
14.147
20.370
1.00
52.80
A


ATOM
64
CB
TYR
A
11
22.683
12.631
20.169
1.00
52.32
A


ATOM
65
CG
TYR
A
11
21.290
12.169
19.791
1.00
51.40
A


ATOM
66
CD1
TYR
A
11
20.788
12.376
18.506
1.00
51.32
A


ATOM
67
CE1
TYR
A
11
19.488
11.996
18.169
1.00
51.46
A


ATOM
68
CD2
TYR
A
11
20.457
11.567
20.732
1.00
50.53
A


ATOM
69
CE2
TYR
A
11
19.158
11.186
20.408
1.00
50.55
A


ATOM
70
CZ
TYR
A
11
18.678
11.403
19.128
1.00
51.02
A


ATOM
71
OH
TYR
A
11
17.387
11.042
18.814
1.00
51.32
A


ATOM
72
C
TYR
A
11
21.882
14.570
21.528
1.00
53.23
A


ATOM
73
O
TYR
A
11
20.705
14.869
21.332
1.00
52.82
A


ATOM
74
N
ALA
A
12
22.440
14.592
22.735
1.00
53.71
A


ATOM
75
CA
ALA
A
12
21.681
14.979
23.923
1.00
53.92
A


ATOM
76
CB
ALA
A
12
22.515
14.745
25.179
1.00
54.54
A


ATOM
77
C
ALA
A
12
21.297
16.447
23.822
1.00
53.28
A


ATOM
78
O
ALA
A
12
20.151
16.826
24.073
1.00
53.07
A


ATOM
79
N
ALA
A
13
22.273
17.267
23.454
1.00
53.23
A


ATOM
80
CA
ALA
A
13
22.062
18.697
23.305
1.00
52.67
A


ATOM
81
CB
ALA
A
13
23.382
19.385
23.012
1.00
52.66
A


ATOM
82
C
ALA
A
13
21.083
18.947
22.170
1.00
52.84
A


ATOM
83
O
ALA
A
13
20.266
19.865
22.239
1.00
53.13
A


ATOM
84
N
MET
A
14
21.164
18.115
21.134
1.00
52.38
A


ATOM
85
CA
MET
A
14
20.295
18.248
19.973
1.00
53.03
A


ATOM
86
CB
MET
A
14
20.715
17.257
18.882
1.00
51.89
A


ATOM
87
CG
MET
A
14
20.349
17.708
17.474
1.00
51.33
A


ATOM
88
SD
MET
A
14
20.752
16.499
16.197
1.00
51.22
A


ATOM
89
CE
MET
A
14
22.491
16.786
15.974
1.00
49.08
A


ATOM
90
C
MET
A
14
18.824
18.042
20.333
1.00
53.97
A


ATOM
91
O
MET
A
14
17.975
18.844
19.960
1.00
54.01
A


ATOM
92
N
GLU
A
15
18.517
16.967
21.053
1.00
55.62
A


ATOM
93
CA
GLU
A
15
17.135
16.710
21.450
1.00
57.23
A


ATOM
94
CB
GLU
A
15
17.037
15.420
22.257
1.00
58.42
A


ATOM
95
CG
GLU
A
15
17.273
14.154
21.472
1.00
59.79
A


ATOM
96
CD
GLU
A
15
17.036
12.922
22.317
1.00
60.46
A


ATOM
97
OE1
GLU
A
15
17.775
12.731
23.307
1.00
60.40
A


ATOM
98
OE2
GLU
A
15
16.106
12.152
21.995
1.00
61.48
A


ATOM
99
C
GLU
A
15
16.621
17.862
22.307
1.00
57.68
A


ATOM
100
O
GLU
A
15
15.477
18.298
22.171
1.00
57.72
A


ATOM
101
N
GLU
A
16
17.482
18.340
23.201
1.00
58.40
A


ATOM
102
CA
GLU
A
16
17.151
19.442
24.094
1.00
59.28
A


ATOM
103
CB
GLU
A
16
18.354
19.746
24.993
1.00
60.79
A


ATOM
104
CG
GLU
A
16
18.104
20.769
26.090
1.00
63.28
A


ATOM
105
CD
GLU
A
16
19.295
20.904
27.027
1.00
65.30
A


ATOM
106
OE1
GLU
A
16
19.675
19.886
27.644
1.00
66.30
A


ATOM
107
OE2
GLU
A
16
19.853
22.019
27.143
1.00
66.14
A


ATOM
108
C
GLU
A
16
16.780
20.673
23.268
1.00
58.94
A


ATOM
109
O
GLU
A
16
15.718
21.272
23.461
1.00
58.58
A


ATOM
110
N
ALA
A
17
17.661
21.041
22.342
1.00
58.27
A


ATOM
111
CA
ALA
A
17
17.422
22.188
21.480
1.00
56.94
A


ATOM
112
CB
ALA
A
17
18.559
22.336
20.484
1.00
57.55
A


ATOM
113
C
ALA
A
17
16.105
21.985
20.748
1.00
56.33
A


ATOM
114
O
ALA
A
17
15.275
22.888
20.676
1.00
56.73
A


ATOM
115
N
ALA
A
18
15.917
20.787
20.208
1.00
55.47
A


ATOM
116
CA
ALA
A
18
14.697
20.466
19.486
1.00
54.97
A


ATOM
117
CB
ALA
A
18
14.796
19.072
18.885
1.00
54.31
A


ATOM
118
C
ALA
A
18
13.511
20.545
20.439
1.00
55.41
A


ATOM
119
O
ALA
A
18
12.408
20.920
20.041
1.00
54.89
A


ATOM
120
N
GLY
A
19
13.749
20.186
21.698
1.00
55.81
A


ATOM
121
CA
GLY
A
19
12.693
20.225
22.690
1.00
56.15
A


ATOM
122
C
GLY
A
19
12.109
21.617
22.812
1.00
56.43
A


ATOM
123
O
GLY
A
19
10.896
21.800
22.726
1.00
57.40
A


ATOM
124
N
LEU
A
20
12.976
22.603
23.011
1.00
56.49
A


ATOM
125
CA
LEU
A
20
12.538
23.985
23.139
1.00
57.06
A


ATOM
126
CB
LEU
A
20
13.747
24.925
23.087
1.00
57.40
A


ATOM
127
CG
LEU
A
20
14.770
24.808
24.223
1.00
56.87
A


ATOM
128
CD1
LEU
A
20
15.963
25.704
23.942
1.00
57.07
A


ATOM
129
CD2
LEU
A
20
14.116
25.196
25.538
1.00
57.91
A


ATOM
130
C
LEU
A
20
11.546
24.355
22.038
1.00
57.57
A


ATOM
131
O
LEU
A
20
10.574
25.069
22.281
1.00
58.18
A


ATOM
132
N
LEU
A
21
11.786
23.853
20.829
1.00
58.02
A


ATOM
133
CA
LEU
A
21
10.916
24.145
19.694
1.00
57.66
A


ATOM
134
CB
LEU
A
21
11.729
24.134
18.399
1.00
58.07
A


ATOM
135
CG
LEU
A
21
12.849
25.173
18.313
1.00
58.73
A


ATOM
136
CD1
LEU
A
21
13.711
24.888
17.097
1.00
59.17
A


ATOM
137
CD2
LEU
A
21
12.252
26.572
18.245
1.00
59.02
A


ATOM
138
C
LEU
A
21
9.749
23.172
19.565
1.00
57.77
A


ATOM
139
O
LEU
A
21
8.904
23.320
18.681
1.00
57.50
A


ATOM
140
N
GLY
A
22
9.702
22.177
20.443
1.00
57.54
A


ATOM
141
CA
GLY
A
22
8.628
21.206
20.386
1.00
56.71
A


ATOM
142
C
GLY
A
22
8.716
20.353
19.136
1.00
56.49
A


ATOM
143
O
GLY
A
22
7.701
19.951
18.564
1.00
55.99
A


ATOM
144
N
VAL
A
23
9.943
20.081
18.707
1.00
56.46
A


ATOM
145
CA
VAL
A
23
10.177
19.267
17.525
1.00
56.41
A


ATOM
146
CB
VAL
A
23
11.229
19.921
16.603
1.00
56.91
A


ATOM
147
CG1
VAL
A
23
11.383
19.104
15.325
1.00
57.54
A


ATOM
148
CG2
VAL
A
23
10.819
21.353
16.279
1.00
56.70
A


ATOM
149
C
VAL
A
23
10.682
17.897
17.967
1.00
56.54
A


ATOM
150
O
VAL
A
23
11.404
17.785
18.958
1.00
56.02
A


ATOM
151
N
ALA
A
24
10.300
16.859
17.231
1.00
56.63
A


ATOM
152
CA
ALA
A
24
10.708
15.498
17.557
1.00
56.80
A


ATOM
153
CB
ALA
A
24
9.568
14.535
17.267
1.00
55.99
A


ATOM
154
C
ALA
A
24
11.949
15.079
16.780
1.00
57.61
A


ATOM
155
O
ALA
A
24
12.082
15.386
15.593
1.00
58.66
A


ATOM
156
N
CYS
A
25
12.857
14.380
17.453
1.00
57.65
A


ATOM
157
CA
CYS
A
25
14.081
13.911
16.817
1.00
58.14
A


ATOM
158
CB
CYS
A
25
15.257
14.002
17.783
1.00
57.35
A


ATOM
159
SG
CYS
A
25
15.899
15.659
17.971
1.00
61.13
A


ATOM
160
C
CYS
A
25
13.949
12.480
16.323
1.00
57.86
A


ATOM
161
O
CYS
A
25
13.127
11.714
16.824
1.00
58.16
A


ATOM
162
N
ALA
A
26
14.768
12.131
15.336
1.00
57.41
A


ATOM
163
CA
ALA
A
26
14.765
10.793
14.759
1.00
57.03
A


ATOM
164
CB
ALA
A
26
14.318
10.854
13.301
1.00
58.04
A


ATOM
165
C
ALA
A
26
16.166
10.208
14.854
1.00
56.71
A


ATOM
166
O
ALA
A
26
17.007
10.454
13.994
1.00
55.93
A


ATOM
167
N
ARG
A
27
16.404
9.431
15.905
1.00
57.35
A


ATOM
168
CA
ARG
A
27
17.703
8.811
16.139
1.00
57.63
A


ATOM
169
CB
ARG
A
27
17.622
7.860
17.332
1.00
58.97
A


ATOM
170
CG
ARG
A
27
18.978
7.432
17.852
1.00
60.53
A


ATOM
171
CD
ARG
A
27
19.022
7.413
19.371
1.00
61.60
A


ATOM
172
NE
ARG
A
27
20.387
7.220
19.851
1.00
62.55
A


ATOM
173
CZ
ARG
A
27
21.098
6.119
19.629
1.00
62.39
A


ATOM
174
NH1
ARG
A
27
20.571
5.121
18.934
1.00
63.03
A


ATOM
175
NH2
ARG
A
27
22.330
6.013
20.106
1.00
63.44
A


ATOM
176
C
ARG
A
27
18.215
8.045
14.926
1.00
57.62
A


ATOM
177
O
ARG
A
27
19.418
8.009
14.667
1.00
57.59
A


ATOM
178
N
ASP
A
28
17.293
7.427
14.193
1.00
57.67
A


ATOM
179
CA
ASP
A
28
17.634
6.653
13.004
1.00
58.11
A


ATOM
180
CB
ASP
A
28
16.369
6.065
12.381
1.00
60.19
A


ATOM
181
CG
ASP
A
28
15.645
5.131
13.316
1.00
62.69
A


ATOM
182
OD1
ASP
A
28
16.187
4.040
13.602
1.00
65.01
A


ATOM
183
OD2
ASP
A
28
14.535
5.492
13.769
1.00
63.34
A


ATOM
184
C
ASP
A
28
18.342
7.495
11.953
1.00
57.11
A


ATOM
185
O
ASP
A
28
19.248
7.019
11.269
1.00
56.40
A


ATOM
186
N
LYS
A
29
17.922
8.750
11.830
1.00
56.47
A


ATOM
187
CA
LYS
A
29
18.494
9.654
10.843
1.00
55.04
A


ATOM
188
CB
LYS
A
29
17.375
10.451
10.172
1.00
55.10
A


ATOM
189
CG
LYS
A
29
16.244
9.581
9.662
1.00
56.13
A


ATOM
190
CD
LYS
A
29
15.225
10.386
8.886
1.00
57.94
A


ATOM
191
CE
LYS
A
29
14.082
9.498
8.434
1.00
58.81
A


ATOM
192
NZ
LYS
A
29
14.581
8.276
7.733
1.00
60.48
A


ATOM
193
C
LYS
A
29
19.531
10.614
11.411
1.00
54.01
A


ATOM
194
O
LYS
A
29
20.078
11.439
10.684
1.00
54.19
A


ATOM
195
N
ILE
A
30
19.809
10.507
12.704
1.00
52.40
A


ATOM
196
CA
ILE
A
30
20.782
11.392
13.323
1.00
51.39
A


ATOM
197
CB
ILE
A
30
20.170
12.124
14.533
1.00
51.07
A


ATOM
198
CG2
ILE
A
30
21.168
13.122
15.096
1.00
51.54
A


ATOM
199
CG1
ILE
A
30
18.887
12.841
14.115
1.00
49.68
A


ATOM
200
CD1
ILE
A
30
19.077
13.829
12.993
1.00
49.27
A


ATOM
201
C
ILE
A
30
22.048
10.676
13.781
1.00
51.27
A


ATOM
202
O
ILE
A
30
23.159
11.111
13.473
1.00
50.40
A


ATOM
203
N
TYR
A
31
21.878
9.573
14.507
1.00
51.61
A


ATOM
204
CA
TYR
A
31
23.018
8.827
15.030
1.00
52.03
A


ATOM
205
CB
TYR
A
31
22.544
7.673
15.918
1.00
54.81
A


ATOM
206
CG
TYR
A
31
23.178
7.688
17.296
1.00
58.36
A


ATOM
207
CD1
TYR
A
31
22.785
8.624
18.253
1.00
60.03
A


ATOM
208
CE1
TYR
A
31
23.381
8.666
19.514
1.00
61.40
A


ATOM
209
CD2
TYR
A
31
24.189
6.788
17.632
1.00
59.71
A


ATOM
210
CE2
TYR
A
31
24.795
6.821
18.891
1.00
61.71
A


ATOM
211
CZ
TYR
A
31
24.384
7.762
19.826
1.00
62.61
A


ATOM
212
OH
TYR
A
31
24.966
7.797
21.074
1.00
63.90
A


ATOM
213
C
TYR
A
31
23.995
8.295
13.989
1.00
50.42
A


ATOM
214
O
TYR
A
31
25.206
8.452
14.141
1.00
50.96
A


ATOM
215
N
PRO
A
32
23.490
7.651
12.921
1.00
49.99
A


ATOM
216
CD
PRO
A
32
22.091
7.335
12.584
1.00
49.46
A


ATOM
217
CA
PRO
A
32
24.401
7.123
11.898
1.00
49.18
A


ATOM
218
CB
PRO
A
32
23.449
6.566
10.841
1.00
48.74
A


ATOM
219
CG
PRO
A
32
22.256
6.163
11.646
1.00
49.30
A


ATOM
220
C
PRO
A
32
25.297
8.222
11.339
1.00
48.53
A


ATOM
221
O
PRO
A
32
26.458
7.984
10.993
1.00
48.53
A


ATOM
222
N
LEU
A
33
24.750
9.430
11.267
1.00
47.26
A


ATOM
223
CA
LEU
A
33
25.486
10.570
10.745
1.00
47.08
A


ATOM
224
CB
LEU
A
33
24.505
11.679
10.356
1.00
46.08
A


ATOM
225
CG
LEU
A
33
25.021
12.774
9.422
1.00
45.61
A


ATOM
226
CD1
LEU
A
33
25.592
12.152
8.156
1.00
44.68
A


ATOM
227
CD2
LEU
A
33
23.875
13.720
9.080
1.00
45.94
A


ATOM
228
C
LEU
A
33
26.517
11.095
11.745
1.00
46.98
A


ATOM
229
O
LEU
A
33
27.673
11.314
11.392
1.00
46.18
A


ATOM
230
N
LEU
A
34
26.104
11.290
12.993
1.00
47.69
A


ATOM
231
CA
LEU
A
34
27.019
11.780
14.021
1.00
49.32
A


ATOM
232
CB
LEU
A
34
26.276
11.978
15.348
1.00
49.84
A


ATOM
233
CG
LEU
A
34
25.193
13.061
15.415
1.00
50.05
A


ATOM
234
CD1
LEU
A
34
24.545
13.051
16.786
1.00
49.62
A


ATOM
235
CD2
LEU
A
34
25.806
14.420
15.136
1.00
50.80
A


ATOM
236
C
LEU
A
34
28.202
10.825
14.229
1.00
49.93
A


ATOM
237
O
LEU
A
34
29.333
11.262
14.445
1.00
50.30
A


ATOM
238
N
SER
A
35
27.936
9.524
14.167
1.00
50.31
A


ATOM
239
CA
SER
A
35
28.984
8.525
14.344
1.00
50.96
A


ATOM
240
CB
SER
A
35
28.395
7.114
14.318
1.00
51.84
A


ATOM
241
OG
SER
A
35
27.562
6.890
15.442
1.00
54.71
A


ATOM
242
C
SER
A
35
30.022
8.652
13.243
1.00
51.29
A


ATOM
243
O
SER
A
35
31.220
8.738
13.512
1.00
51.44
A


ATOM
244
N
THR
A
36
29.554
8.669
12.000
1.00
50.28
A


ATOM
245
CA
THR
A
36
30.444
8.782
10.855
1.00
49.52
A


ATOM
246
CB
THR
A
36
29.645
8.980
9.559
1.00
49.26
A


ATOM
247
OG1
THR
A
36
28.690
7.923
9.425
1.00
49.50
A


ATOM
248
CG2
THR
A
36
30.568
8.968
8.358
1.00
49.15
A


ATOM
249
C
THR
A
36
31.410
9.950
11.025
1.00
49.71
A


ATOM
250
O
THR
A
36
32.552
9.893
10.571
1.00
50.28
A


ATOM
251
N
PHE
A
37
30.956
11.009
11.689
1.00
49.29
A


ATOM
252
CA
PHE
A
37
31.807
12.174
11.891
1.00
48.84
A


ATOM
253
CB
PHE
A
37
31.146
13.416
11.284
1.00
46.61
A


ATOM
254
CG
PHE
A
37
30.934
13.323
9.799
1.00
43.07
A


ATOM
255
CD1
PHE
A
37
29.729
12.873
9.283
1.00
41.56
A


ATOM
256
CD2
PHE
A
37
31.957
13.656
8.920
1.00
39.81
A


ATOM
257
CE1
PHE
A
37
29.543
12.754
7.908
1.00
42.57
A


ATOM
258
CE2
PHE
A
37
31.785
13.542
7.549
1.00
41.01
A


ATOM
259
CZ
PHE
A
37
30.576
13.090
7.039
1.00
40.00
A


ATOM
260
C
PHE
A
37
32.136
12.423
13.359
1.00
50.82
A


ATOM
261
O
PHE
A
37
32.304
13.568
13.781
1.00
50.54
A


ATOM
262
N
GLN
A
38
32.246
11.343
14.128
1.00
52.93
A


ATOM
263
CA
GLN
A
38
32.543
11.433
15.557
1.00
54.33
A


ATOM
264
CB
GLN
A
38
32.543
10.036
16.180
1.00
55.97
A


ATOM
265
CG
GLN
A
38
33.536
9.088
15.535
1.00
58.11
A


ATOM
266
CD
GLN
A
38
33.584
7.741
16.221
1.00
59.72
A


ATOM
267
OE1
GLN
A
38
32.549
7.124
16.483
1.00
60.66
A


ATOM
268
NE2
GLN
A
38
34.790
7.271
16.510
1.00
60.33
A


ATOM
269
C
GLN
A
38
33.871
12.113
15.870
1.00
53.93
A


ATOM
270
O
GLN
A
38
33.990
12.818
16.868
1.00
53.58
A


ATOM
271
N
ASP
A
39
34.868
11.901
15.022
1.00
53.91
A


ATOM
272
CA
ASP
A
39
36.178
12.495
15.252
1.00
54.34
A


ATOM
273
CB
ASP
A
39
37.199
11.935
14.259
1.00
55.58
A


ATOM
274
CG
ASP
A
39
37.421
10.443
14.431
1.00
57.29
A


ATOM
275
OD1
ASP
A
39
37.553
9.990
15.587
1.00
57.11
A


ATOM
276
OD2
ASP
A
39
37.475
9.724
13.409
1.00
58.78
A


ATOM
277
C
ASP
A
39
36.188
14.019
15.186
1.00
54.75
A


ATOM
278
O
ASP
A
39
37.163
14.653
15.594
1.00
53.65
A


ATOM
279
N
THR
A
40
35.104
14.608
14.686
1.00
54.95
A


ATOM
280
CA
THR
A
40
35.019
16.061
14.569
1.00
55.03
A


ATOM
281
CB
THR
A
40
34.192
16.487
13.336
1.00
53.61
A


ATOM
282
OG1
THR
A
40
32.825
16.109
13.526
1.00
51.16
A


ATOM
283
CG2
THR
A
40
34.720
15.833
12.080
1.00
52.82
A


ATOM
284
C
THR
A
40
34.366
16.707
15.783
1.00
56.50
A


ATOM
285
O
THR
A
40
34.613
17.876
16.082
1.00
57.15
A


ATOM
286
N
LEU
A
41
33.536
15.937
16.478
1.00
57.42
A


ATOM
287
CA
LEU
A
41
32.803
16.432
17.639
1.00
58.80
A


ATOM
288
CB
LEU
A
41
31.702
15.430
18.004
1.00
57.63
A


ATOM
289
CG
LEU
A
41
30.761
15.083
16.844
1.00
56.86
A


ATOM
290
CD1
LEU
A
41
29.770
14.018
17.277
1.00
55.61
A


ATOM
291
CD2
LEU
A
41
30.039
16.341
16.378
1.00
56.19
A


ATOM
292
C
LEU
A
41
33.647
16.751
18.871
1.00
60.36
A


ATOM
293
O
LEU
A
41
33.108
17.030
19.942
1.00
60.30
A


ATOM
294
N
VAL
A
42
34.967
16.725
18.719
1.00
62.05
A


ATOM
295
CA
VAL
A
42
35.859
17.015
19.834
1.00
63.74
A


ATOM
296
CB
VAL
A
42
37.280
16.494
19.562
1.00
64.26
A


ATOM
297
CG1
VAL
A
42
38.190
16.837
20.733
1.00
65.42
A


ATOM
298
CG2
VAL
A
42
37.243
14.990
19.333
1.00
64.88
A


ATOM
299
C
VAL
A
42
35.942
18.510
20.126
1.00
64.12
A


ATOM
300
O
VAL
A
42
36.493
19.277
19.332
1.00
64.30
A


ATOM
301
N
GLU
A
43
35.398
18.910
21.273
1.00
64.21
A


ATOM
302
CA
GLU
A
43
35.403
20.308
21.694
1.00
64.09
A


ATOM
303
CB
GLU
A
43
35.025
20.410
23.175
1.00
65.49
A


ATOM
304
CG
GLU
A
43
33.621
19.924
23.503
1.00
68.00
A


ATOM
305
CD
GLU
A
43
33.338
19.927
24.997
1.00
69.13
A


ATOM
306
OE1
GLU
A
43
34.017
19.180
25.733
1.00
68.77
A


ATOM
307
OE2
GLU
A
43
32.440
20.677
25.436
1.00
70.84
A


ATOM
308
C
GLU
A
43
36.770
20.955
21.473
1.00
63.27
A


ATOM
309
O
GLU
A
43
37.795
20.424
21.901
1.00
63.43
A


ATOM
310
N
GLY
A
44
36.778
22.102
20.802
1.00
61.60
A


ATOM
311
CA
GLY
A
44
38.027
22.792
20.543
1.00
59.61
A


ATOM
312
C
GLY
A
44
38.003
23.516
19.215
1.00
58.39
A


ATOM
313
O
GLY
A
44
37.805
24.730
19.161
1.00
59.02
A


ATOM
314
N
GLY
A
45
38.219
22.770
18.138
1.00
56.07
A


ATOM
315
CA
GLY
A
45
38.201
23.362
16.816
1.00
54.08
A


ATOM
316
C
GLY
A
45
36.946
22.906
16.101
1.00
52.48
A


ATOM
317
O
GLY
A
45
36.914
22.817
14.879
1.00
52.24
A


ATOM
318
N
SER
A
46
35.911
22.613
16.883
1.00
51.11
A


ATOM
319
CA
SER
A
46
34.633
22.152
16.358
1.00
48.42
A


ATOM
320
CB
SER
A
46
34.175
20.912
17.122
1.00
48.98
A


ATOM
321
OG
SER
A
46
32.801
20.655
16.893
1.00
49.29
A


ATOM
322
C
SER
A
46
33.531
23.203
16.423
1.00
47.59
A


ATOM
323
O
SER
A
46
33.421
23.960
17.391
1.00
46.05
A


ATOM
324
N
VAL
A
47
32.711
23.243
15.380
1.00
45.49
A


ATOM
325
CA
VAL
A
47
31.605
24.181
15.326
1.00
42.26
A


ATOM
326
CB
VAL
A
47
31.764
25.186
14.164
1.00
41.88
A


ATOM
327
CG1
VAL
A
47
30.602
26.174
14.167
1.00
41.99
A


ATOM
328
CG2
VAL
A
47
33.079
25.937
14.295
1.00
41.85
A


ATOM
329
C
VAL
A
47
30.323
23.390
15.129
1.00
41.67
A


ATOM
330
O
VAL
A
47
30.199
22.631
14.171
1.00
42.19
A


ATOM
331
N
VAL
A
48
29.388
23.536
16.059
1.00
40.43
A


ATOM
332
CA
VAL
A
48
28.111
22.849
15.962
1.00
39.58
A


ATOM
333
CB
VAL
A
48
27.974
21.711
16.995
1.00
40.32
A


ATOM
334
CG1
VAL
A
48
26.527
21.222
17.041
1.00
39.82
A


ATOM
335
CG2
VAL
A
48
28.889
20.553
16.619
1.00
40.14
A


ATOM
336
C
VAL
A
48
27.012
23.869
16.197
1.00
39.83
A


ATOM
337
O
VAL
A
48
27.061
24.636
17.159
1.00
39.49
A


ATOM
338
N
VAL
A
49
26.023
23.874
15.308
1.00
39.22
A


ATOM
339
CA
VAL
A
49
24.914
24.808
15.393
1.00
38.77
A


ATOM
340
CB
VAL
A
49
24.974
25.838
14.235
1.00
38.65
A


ATOM
341
CG1
VAL
A
49
24.027
26.994
14.514
1.00
39.77
A


ATOM
342
CG2
VAL
A
49
26.394
26.331
14.047
1.00
37.33
A


ATOM
343
C
VAL
A
49
23.571
24.092
15.314
1.00
39.16
A


ATOM
344
O
VAL
A
49
23.427
23.091
14.611
1.00
39.92
A


ATOM
345
N
PHE
A
50
22.596
24.601
16.057
1.00
38.97
A


ATOM
346
CA
PHE
A
50
21.243
24.061
16.038
1.00
39.87
A


ATOM
347
CB
PHE
A
50
20.834
23.575
17.425
1.00
39.82
A


ATOM
348
CG
PHE
A
50
21.720
22.493
17.971
1.00
41.79
A


ATOM
349
CD1
PHE
A
50
22.076
21.400
17.182
1.00
42.66
A


ATOM
350
CD2
PHE
A
50
22.167
22.544
19.288
1.00
41.26
A


ATOM
351
CE1
PHE
A
50
22.864
20.368
17.699
1.00
43.88
A


ATOM
352
CE2
PHE
A
50
22.953
21.520
19.814
1.00
42.58
A


ATOM
353
CZ
PHE
A
50
23.302
20.429
19.019
1.00
41.73
A


ATOM
354
C
PHE
A
50
20.380
25.242
15.598
1.00
39.56
A


ATOM
355
O
PHE
A
50
20.261
26.227
16.320
1.00
38.63
A


ATOM
356
N
SER
A
51
19.772
25.139
14.419
1.00
39.59
A


ATOM
357
CA
SER
A
51
19.002
26.256
13.892
1.00
38.98
A


ATOM
358
CB
SER
A
51
19.631
26.692
12.571
1.00
39.41
A


ATOM
359
OG
SER
A
51
21.022
26.893
12.739
1.00
40.57
A


ATOM
360
C
SER
A
51
17.500
26.086
13.706
1.00
39.41
A


ATOM
361
O
SER
A
51
17.018
25.037
13.283
1.00
38.55
A


ATOM
362
N
MET
A
52
16.778
27.163
14.006
1.00
39.56
A


ATOM
363
CA
MET
A
52
15.326
27.203
13.904
1.00
41.09
A


ATOM
364
CB
MET
A
52
14.721
27.612
15.248
1.00
42.98
A


ATOM
365
CG
MET
A
52
14.952
29.080
15.553
1.00
44.79
A


ATOM
366
SD
MET
A
52
14.137
29.694
17.032
1.00
53.36
A


ATOM
367
CE
MET
A
52
12.461
29.878
16.420
1.00
50.19
A


ATOM
368
C
MET
A
52
14.948
28.254
12.867
1.00
39.86
A


ATOM
369
O
MET
A
52
15.736
29.144
12.577
1.00
37.96
A


ATOM
370
N
ALA
A
53
13.735
28.155
12.329
1.00
41.96
A


ATOM
371
CA
ALA
A
53
13.240
29.115
11.344
1.00
43.31
A


ATOM
372
CB
ALA
A
53
13.443
28.583
9.930
1.00
43.60
A


ATOM
373
C
ALA
A
53
11.757
29.373
11.605
1.00
44.82
A


ATOM
374
O
ALA
A
53
11.035
28.486
12.058
1.00
45.88
A


ATOM
375
N
SER
A
54
11.309
30.588
11.314
1.00
45.31
A


ATOM
376
CA
SER
A
54
9.920
30.975
11.536
1.00
46.31
A


ATOM
377
CB
SER
A
54
9.824
32.490
11.704
1.00
44.93
A


ATOM
378
OG
SER
A
54
9.947
33.126
10.439
1.00
42.54
A


ATOM
379
C
SER
A
54
8.992
30.569
10.402
1.00
47.54
A


ATOM
380
O
SER
A
54
9.417
30.002
9.398
1.00
47.54
A


ATOM
381
N
GLY
A
55
7.713
30.889
10.583
1.00
50.46
A


ATOM
382
CA
GLY
A
55
6.701
30.596
9.585
1.00
52.05
A


ATOM
383
C
GLY
A
55
6.526
29.136
9.219
1.00
52.73
A


ATOM
384
O
GLY
A
55
6.502
28.256
10.081
1.00
52.54
A


ATOM
385
N
ARG
A
56
6.399
28.887
7.922
1.00
53.50
A


ATOM
386
CA
ARG
A
56
6.206
27.539
7.408
1.00
54.78
A


ATOM
387
CB
ARG
A
56
5.885
27.590
5.911
1.00
55.54
A


ATOM
388
CG
ARG
A
56
7.112
27.702
5.021
1.00
55.10
A


ATOM
389
CD
ARG
A
56
6.731
27.742
3.551
1.00
56.42
A


ATOM
390
NE
ARG
A
56
7.892
27.536
2.690
1.00
58.46
A


ATOM
391
CZ
ARG
A
56
7.858
27.590
1.363
1.00
59.31
A


ATOM
392
NH1
ARG
A
56
6.718
27.849
0.737
1.00
59.88
A


ATOM
393
NH2
ARG
A
56
8.961
27.375
0.658
1.00
59.39
A


ATOM
394
C
ARG
A
56
7.438
26.666
7.626
1.00
54.74
A


ATOM
395
O
ARG
A
56
7.404
25.464
7.359
1.00
54.18
A


ATOM
396
N
HIS
A
57
8.522
27.270
8.105
1.00
54.51
A


ATOM
397
CA
HIS
A
57
9.759
26.525
8.335
1.00
54.29
A


ATOM
398
CB
HIS
A
57
10.964
27.302
7.779
1.00
55.16
A


ATOM
399
CG
HIS
A
57
10.719
27.936
6.444
1.00
56.03
A


ATOM
400
CD2
HIS
A
57
11.050
27.537
5.193
1.00
56.33
A


ATOM
401
ND1
HIS
A
57
10.032
29.122
6.298
1.00
56.66
A


ATOM
402
CE1
HIS
A
57
9.948
29.426
5.014
1.00
56.21
A


ATOM
403
NE2
HIS
A
57
10.557
28.480
4.322
1.00
56.40
A


ATOM
404
C
HIS
A
57
9.986
26.266
9.822
1.00
53.88
A


ATOM
405
O
HIS
A
57
11.051
25.791
10.220
1.00
53.33
A


ATOM
406
N
SER
A
58
8.980
26.559
10.639
1.00
53.09
A


ATOM
407
CA
SER
A
58
9.108
26.402
12.085
1.00
52.87
A


ATOM
408
CB
SER
A
58
8.133
27.346
12.794
1.00
53.36
A


ATOM
409
OG
SER
A
58
6.792
27.058
12.437
1.00
54.09
A


ATOM
410
C
SER
A
58
8.967
25.009
12.689
1.00
51.52
A


ATOM
411
O
SER
A
58
9.114
24.859
13.901
1.00
52.42
A


ATOM
412
N
THR
A
59
8.696
23.995
11.874
1.00
49.99
A


ATOM
413
CA
THR
A
59
8.540
22.644
12.408
1.00
49.61
A


ATOM
414
CB
THR
A
59
7.321
21.926
11.805
1.00
50.26
A


ATOM
415
OG1
THR
A
59
7.565
21.660
10.418
1.00
52.05
A


ATOM
416
CG2
THR
A
59
6.070
22.778
11.955
1.00
49.84
A


ATOM
417
C
THR
A
59
9.750
21.755
12.168
1.00
48.80
A


ATOM
418
O
THR
A
59
9.627
20.534
12.141
1.00
50.12
A


ATOM
419
N
GLU
A
60
10.917
22.359
11.996
1.00
47.87
A


ATOM
420
CA
GLU
A
60
12.127
21.583
11.760
1.00
48.24
A


ATOM
421
CB
GLU
A
60
12.428
21.520
10.260
1.00
49.44
A


ATOM
422
CG
GLU
A
60
11.638
20.460
9.520
1.00
53.29
A


ATOM
423
CD
GLU
A
60
11.520
20.742
8.030
1.00
56.50
A


ATOM
424
OE1
GLU
A
60
12.551
21.045
7.391
1.00
55.69
A


ATOM
425
OE2
GLU
A
60
10.389
20.657
7.497
1.00
59.38
A


ATOM
426
C
GLU
A
60
13.328
22.148
12.498
1.00
46.12
A


ATOM
427
O
GLU
A
60
13.344
23.319
12.886
1.00
46.85
A


ATOM
428
N
LEU
A
61
14.328
21.301
12.708
1.00
43.44
A


ATOM
429
CA
LEU
A
61
15.544
21.737
13.374
1.00
41.98
A


ATOM
430
CB
LEU
A
61
15.603
21.245
14.822
1.00
41.96
A


ATOM
431
CG
LEU
A
61
16.830
21.736
15.603
1.00
40.28
A


ATOM
432
CD1
LEU
A
61
16.748
23.234
15.798
1.00
41.49
A


ATOM
433
CD2
LEU
A
61
16.899
21.043
16.957
1.00
43.82
A


ATOM
434
C
LEU
A
61
16.748
21.216
12.613
1.00
40.23
A


ATOM
435
O
LEU
A
61
16.951
20.005
12.487
1.00
38.93
A


ATOM
436
N
ASP
A
62
17.535
22.153
12.100
1.00
39.30
A


ATOM
437
CA
ASP
A
62
18.736
21.840
11.348
1.00
38.26
A


ATOM
438
CB
ASP
A
62
18.972
22.893
10.263
1.00
37.02
A


ATOM
439
CG
ASP
A
62
18.188
22.617
8.993
1.00
38.78
A


ATOM
440
OD1
ASP
A
62
17.534
21.557
8.915
1.00
38.35
A


ATOM
441
OD2
ASP
A
62
18.234
23.459
8.069
1.00
35.53
A


ATOM
442
C
ASP
A
62
19.948
21.817
12.261
1.00
37.32
A


ATOM
443
O
ASP
A
62
20.004
22.555
13.251
1.00
38.00
A


ATOM
444
N
PHE
A
63
20.905
20.956
11.938
1.00
36.34
A


ATOM
445
CA
PHE
A
63
22.142
20.894
12.692
1.00
36.73
A


ATOM
446
CB
PHE
A
63
22.235
19.624
13.552
1.00
38.45
A


ATOM
447
CG
PHE
A
63
22.102
18.336
12.790
1.00
40.09
A


ATOM
448
CD1
PHE
A
63
20.849
17.814
12.491
1.00
41.21
A


ATOM
449
CD2
PHE
A
63
23.230
17.620
12.413
1.00
38.85
A


ATOM
450
CE1
PHE
A
63
20.722
16.596
11.830
1.00
41.80
A


ATOM
451
CE2
PHE
A
63
23.117
16.405
11.753
1.00
40.48
A


ATOM
452
CZ
PHE
A
63
21.857
15.889
11.460
1.00
41.84
A


ATOM
453
C
PHE
A
63
23.297
20.961
11.709
1.00
36.67
A


ATOM
454
O
PHE
A
63
23.337
20.215
10.724
1.00
37.63
A


ATOM
455
N
SER
A
64
24.217
21.884
11.966
1.00
37.08
A


ATOM
456
CA
SER
A
64
25.389
22.078
11.123
1.00
37.11
A


ATOM
457
CB
SER
A
64
25.533
23.550
10.742
1.00
37.02
A


ATOM
458
OG
SER
A
64
24.347
24.054
10.156
1.00
37.59
A


ATOM
459
C
SER
A
64
26.617
21.641
11.908
1.00
37.78
A


ATOM
460
O
SER
A
64
26.776
22.004
13.070
1.00
38.14
A


ATOM
461
N
ILE
A
65
27.486
20.869
11.269
1.00
37.84
A


ATOM
462
CA
ILE
A
65
28.687
20.375
11.931
1.00
38.20
A


ATOM
463
CB
ILE
A
65
28.517
18.881
12.298
1.00
38.18
A


ATOM
464
CG2
ILE
A
65
29.813
18.319
12.870
1.00
39.41
A


ATOM
465
CG1
ILE
A
65
27.372
18.732
13.306
1.00
37.70
A


ATOM
466
CD1
ILE
A
65
26.947
17.304
13.552
1.00
38.47
A


ATOM
467
C
ILE
A
65
29.918
20.549
11.053
1.00
38.20
A


ATOM
468
O
ILE
A
65
29.936
20.120
9.903
1.00
38.89
A


ATOM
469
N
SER
A
66
30.947
21.184
11.598
1.00
38.03
A


ATOM
470
CA
SER
A
66
32.178
21.396
10.848
1.00
38.52
A


ATOM
471
CB
SER
A
66
33.091
22.367
11.599
1.00
37.05
A


ATOM
472
OG
SER
A
66
33.302
21.907
12.920
1.00
39.76
A


ATOM
473
C
SER
A
66
32.897
20.067
10.652
1.00
38.47
A


ATOM
474
O
SER
A
66
32.773
19.162
11.474
1.00
39.22
A


ATOM
475
N
VAL
A
67
33.635
19.954
9.553
1.00
37.19
A


ATOM
476
CA
VAL
A
67
34.393
18.745
9.242
1.00
36.16
A


ATOM
477
CB
VAL
A
67
33.639
17.821
8.251
1.00
36.78
A


ATOM
478
CG1
VAL
A
67
34.466
16.561
7.977
1.00
34.33
A


ATOM
479
CG2
VAL
A
67
32.277
17.439
8.816
1.00
33.97
A


ATOM
480
C
VAL
A
67
35.715
19.174
8.604
1.00
36.50
A


ATOM
481
O
VAL
A
67
35.739
19.707
7.496
1.00
34.41
A


ATOM
482
N
PRO
A
68
36.834
18.970
9.315
1.00
38.13
A


ATOM
483
CD
PRO
A
68
36.945
18.356
10.650
1.00
38.47
A


ATOM
484
CA
PRO
A
68
38.156
19.346
8.795
1.00
38.92
A


ATOM
485
CB
PRO
A
68
39.104
18.943
9.927
1.00
39.16
A


ATOM
486
CG
PRO
A
68
38.355
17.833
10.630
1.00
38.66
A


ATOM
487
C
PRO
A
68
38.469
18.638
7.482
1.00
38.57
A


ATOM
488
O
PRO
A
68
38.045
17.506
7.274
1.00
39.03
A


ATOM
489
N
THR
A
69
39.200
19.309
6.598
1.00
40.67
A


ATOM
490
CA
THR
A
69
39.542
18.736
5.297
1.00
41.89
A


ATOM
491
CB
THR
A
69
40.363
19.719
4.449
1.00
41.81
A


ATOM
492
OG1
THR
A
69
41.533
20.109
5.174
1.00
45.53
A


ATOM
493
CG2
THR
A
69
39.548
20.949
4.115
1.00
41.08
A


ATOM
494
C
THR
A
69
40.335
17.443
5.420
1.00
42.77
A


ATOM
495
O
THR
A
69
40.345
16.619
4.499
1.00
40.61
A


ATOM
496
N
SER
A
70
40.992
17.270
6.563
1.00
43.04
A


ATOM
497
CA
SER
A
70
41.802
16.083
6.810
1.00
44.22
A


ATOM
498
CB
SER
A
70
42.565
16.231
8.133
1.00
44.13
A


ATOM
499
OG
SER
A
70
41.679
16.254
9.239
1.00
42.16
A


ATOM
500
C
SER
A
70
40.934
14.833
6.854
1.00
44.81
A


ATOM
501
O
SER
A
70
41.387
13.734
6.531
1.00
45.00
A


ATOM
502
N
HIS
A
71
39.679
15.000
7.250
1.00
44.99
A


ATOM
503
CA
HIS
A
71
38.779
13.866
7.328
1.00
45.50
A


ATOM
504
CB
HIS
A
71
37.595
14.215
8.230
1.00
50.25
A


ATOM
505
CG
HIS
A
71
37.970
14.374
9.671
1.00
54.93
A


ATOM
506
CD2
HIS
A
71
39.181
14.404
10.279
1.00
56.60
A


ATOM
507
ND1
HIS
A
71
37.036
14.525
10.674
1.00
56.61
A


ATOM
508
CE1
HIS
A
71
37.656
14.640
11.837
1.00
57.31
A


ATOM
509
NE2
HIS
A
71
38.957
14.570
11.625
1.00
57.50
A


ATOM
510
C
HIS
A
71
38.313
13.397
5.948
1.00
44.27
A


ATOM
511
O
HIS
A
71
37.778
12.299
5.803
1.00
42.77
A


ATOM
512
N
GLY
A
72
38.539
14.226
4.935
1.00
42.93
A


ATOM
513
CA
GLY
A
72
38.151
13.868
3.582
1.00
42.01
A


ATOM
514
C
GLY
A
72
36.825
14.454
3.136
1.00
40.53
A


ATOM
515
O
GLY
A
72
36.046
14.934
3.954
1.00
40.70
A


ATOM
516
N
ASP
A
73
36.581
14.423
1.828
1.00
39.45
A


ATOM
517
CA
ASP
A
73
35.345
14.931
1.238
1.00
39.65
A


ATOM
518
CB
ASP
A
73
35.309
14.568
−0.251
1.00
39.15
A


ATOM
519
CG
ASP
A
73
34.036
15.025
−0.942
1.00
37.87
A


ATOM
520
OD1
ASP
A
73
32.947
14.913
−0.343
1.00
39.76
A


ATOM
521
OD2
ASP
A
73
34.122
15.485
−2.097
1.00
40.00
A


ATOM
522
C
ASP
A
73
34.162
14.280
1.957
1.00
37.48
A


ATOM
523
O
ASP
A
73
33.993
13.066
1.895
1.00
38.41
A


ATOM
524
N
PRO
A
74
33.335
15.079
2.652
1.00
36.55
A


ATOM
525
CD
PRO
A
74
33.529
16.507
2.958
1.00
37.15
A


ATOM
526
CA
PRO
A
74
32.175
14.544
3.378
1.00
34.86
A


ATOM
527
CB
PRO
A
74
31.752
15.714
4.268
1.00
34.11
A


ATOM
528
CG
PRO
A
74
32.170
16.906
3.466
1.00
36.59
A


ATOM
529
C
PRO
A
74
31.027
14.004
2.518
1.00
33.80
A


ATOM
530
O
PRO
A
74
30.274
13.156
2.982
1.00
33.04
A


ATOM
531
N
TYR
A
75
30.867
14.494
1.289
1.00
32.38
A


ATOM
532
CA
TYR
A
75
29.801
13.957
0.447
1.00
32.91
A


ATOM
533
CD
TYR
A
75
29.530
14.819
−0.789
1.00
32.39
A


ATOM
534
CG
TYR
A
75
28.334
14.320
−1.600
1.00
32.65
A


ATOM
535
CD1
TYR
A
75
27.086
14.159
−1.002
1.00
32.34
A


ATOM
536
CE1
TYR
A
75
25.978
13.752
−1.731
1.00
32.78
A


ATOM
537
CD2
TYR
A
75
28.442
14.047
−2.966
1.00
35.34
A


ATOM
538
CE2
TYR
A
75
27.321
13.632
−3.714
1.00
33.58
A


ATOM
539
CZ
TYR
A
75
26.098
13.494
−3.084
1.00
33.86
A


ATOM
540
OH
TYR
A
75
24.972
13.132
−3.797
1.00
33.63
A


ATOM
541
C
TYR
A
75
30.256
12.580
−0.007
1.00
33.77
A


ATOM
542
O
TYR
A
75
29.464
11.641
−0.055
1.00
33.18
A


ATOM
543
N
ALA
A
76
31.540
12.470
−0.346
1.00
34.39
A


ATOM
544
CA
ALA
A
76
32.098
11.187
−0.772
1.00
35.15
A


ATOM
545
CB
ALA
A
76
33.595
11.319
−1.071
1.00
34.26
A


ATOM
546
C
ALA
A
76
31.885
10.199
0.362
1.00
34.76
A


ATOM
547
O
ALA
A
76
31.608
9.023
0.136
1.00
33.20
A


ATOM
548
N
THR
A
77
32.005
10.695
1.588
1.00
34.77
A


ATOM
549
CA
THR
A
77
31.824
9.858
2.767
1.00
36.06
A


ATOM
550
CB
THR
A
77
32.317
10.584
4.034
1.00
36.33
A


ATOM
551
OG1
THR
A
77
33.740
10.719
3.968
1.00
37.57
A


ATOM
552
CG2
THR
A
77
31.941
9.798
5.289
1.00
36.53
A


ATOM
553
C
THR
A
77
30.385
9.390
2.999
1.00
37.01
A


ATOM
554
O
THR
A
77
30.152
8.203
3.263
1.00
38.11
A


ATOM
555
N
VAL
A
78
29.420
10.303
2.911
1.00
36.63
A


ATOM
556
CA
VAL
A
78
28.027
9.923
3.138
1.00
36.66
A


ATOM
557
CB
VAL
A
78
27.087
11.164
3.268
1.00
36.95
A


ATOM
558
CG1
VAL
A
78
27.499
11.998
4.468
1.00
36.05
A


ATOM
559
CG2
VAL
A
78
27.124
12.002
1.999
1.00
36.59
A


ATOM
560
C
VAL
A
78
27.480
9.003
2.053
1.00
35.80
A


ATOM
561
O
VAL
A
78
26.614
8.176
2.325
1.00
35.84
A


ATOM
562
N
VAL
A
79
27.972
9.135
0.826
1.00
35.93
A


ATOM
563
CA
VAL
A
79
27.485
8.269
−0.241
1.00
36.96
A


ATOM
564
CB
VAL
A
79
27.857
8.808
−1.641
1.00
36.08
A


ATOM
565
CG1
VAL
A
79
27.323
7.859
−2.712
1.00
34.68
A


ATOM
566
CG2
VAL
A
79
27.278
10.197
−1.844
1.00
36.45
A


ATOM
567
C
VAL
A
79
28.091
6.872
−0.064
1.00
37.92
A


ATOM
568
O
VAL
A
79
27.384
5.867
−0.111
1.00
38.87
A


ATOM
569
N
GLU
A
80
29.402
6.823
0.155
1.00
38.87
A


ATOM
570
CA
GLU
A
80
30.106
5.562
0.345
1.00
40.08
A


ATOM
571
CB
GLU
A
80
31.589
5.809
0.592
1.00
42.04
A


ATOM
572
CG
GLU
A
80
32.435
5.911
−0.653
1.00
47.22
A


ATOM
573
CD
GLU
A
80
33.913
5.949
−0.316
1.00
50.49
A


ATOM
574
OE1
GLU
A
80
34.379
5.027
0.396
1.00
51.53
A


ATOM
575
OE2
GLU
A
80
34.603
6.895
−0.760
1.00
52.54
A


ATOM
576
C
GLU
A
80
29.579
4.732
1.502
1.00
40.32
A


ATOM
577
O
GLU
A
80
29.569
3.504
1.436
1.00
39.33
A


ATOM
578
N
LYS
A
81
29.158
5.401
2.567
1.00
40.75
A


ATOM
579
CA
LYS
A
81
28.658
4.701
3.743
1.00
42.50
A


ATOM
580
CB
LYS
A
81
28.979
5.515
4.999
1.00
43.97
A


ATOM
581
CG
LYS
A
81
28.806
4.743
6.294
1.00
47.62
A


ATOM
582
CD
LYS
A
81
29.508
5.437
7.448
1.00
48.42
A


ATOM
583
CE
LYS
A
81
29.424
4.611
8.722
1.00
49.73
A


ATOM
584
NZ
LYS
A
81
30.230
5.215
9.816
1.00
50.46
A


ATOM
585
C
LYS
A
81
27.160
4.393
3.659
1.00
42.43
A


ATOM
586
O
LYS
A
81
26.583
3.794
4.569
1.00
43.82
A


ATOM
587
N
GLY
A
82
26.534
4.800
2.561
1.00
41.47
A


ATOM
588
CA
GLY
A
82
25.120
4.530
2.380
1.00
40.57
A


ATOM
589
C
GLY
A
82
24.167
5.412
3.165
1.00
39.41
A


ATOM
590
O
GLY
A
82
23.027
5.017
3.410
1.00
39.15
A


ATOM
591
N
LEU
A
83
24.617
6.606
3.551
1.00
38.67
A


ATOM
592
CA
LEU
A
83
23.776
7.530
4.310
1.00
36.40
A


ATOM
593
CB
LEU
A
83
24.625
8.331
5.296
1.00
37.66
A


ATOM
594
CG
LEU
A
83
25.301
7.524
6.409
1.00
39.03
A


ATOM
595
CD1
LEU
A
83
26.040
8.469
7.328
1.00
39.46
A


ATOM
596
CD2
LEU
A
83
24.260
6.734
7.203
1.00
38.72
A


ATOM
597
C
LEU
A
83
22.999
8.481
3.407
1.00
36.46
A


ATOM
598
O
LEU
A
83
22.058
9.144
3.845
1.00
38.34
A


ATOM
599
N
PHE
A
84
23.397
8.569
2.145
1.00
33.71
A


ATOM
600
CA
PHE
A
84
22.695
9.427
1.206
1.00
34.20
A


ATOM
601
CB
PHE
A
84
23.172
10.885
1.305
1.00
35.21
A


ATOM
602
CG
PHE
A
84
22.307
11.840
0.541
1.00
33.04
A


ATOM
603
CD1
PHE
A
84
21.109
12.290
1.080
1.00
33.86
A


ATOM
604
CD2
PHE
A
84
22.636
12.211
−0.758
1.00
32.80
A


ATOM
605
CE1
PHE
A
84
20.247
13.088
0.335
1.00
33.51
A


ATOM
606
CE2
PHE
A
84
21.780
13.010
−1.513
1.00
32.34
A


ATOM
607
CZ
PHE
A
84
20.583
13.449
−0.970
1.00
33.64
A


ATOM
608
C
PHE
A
84
22.952
8.909
−0.199
1.00
34.14
A


ATOM
609
O
PHE
A
84
24.055
8.456
−0.502
1.00
32.87
A


ATOM
610
N
PRO
A
85
21.936
8.965
−1.075
1.00
35.40
A


ATOM
611
CD
PRO
A
85
20.507
9.187
−0.785
1.00
35.87
A


ATOM
612
CA
PRO
A
85
22.115
8.481
−2.446
1.00
36.99
A


ATOM
613
CB
PRO
A
85
20.743
7.912
−2.782
1.00
36.41
A


ATOM
614
CG
PRO
A
85
19.836
8.885
−2.131
1.00
36.37
A


ATOM
615
C
PRO
A
85
22.542
9.560
−3.432
1.00
38.58
A


ATOM
616
O
PRO
A
85
22.088
10.697
−3.343
1.00
37.08
A


ATOM
617
N
ALA
A
86
23.423
9.204
−4.365
1.00
41.13
A


ATOM
618
CA
ALA
A
86
23.854
10.154
−5.378
1.00
43.66
A


ATOM
619
CB
ALA
A
86
24.889
9.528
−6.295
1.00
43.21
A


ATOM
620
C
ALA
A
86
22.605
10.524
−6.171
1.00
45.42
A


ATOM
621
O
ALA
A
86
21.847
9.649
−6.600
1.00
45.70
A


ATOM
622
N
THR
A
87
22.389
11.821
−6.355
1.00
46.96
A


ATOM
623
CA
THR
A
87
21.222
12.308
−7.076
1.00
47.91
A


ATOM
624
CB
THR
A
87
21.011
13.810
−6.820
1.00
49.25
A


ATOM
625
OG1
THR
A
87
22.142
14.537
−7.318
1.00
50.27
A


ATOM
626
CG2
THR
A
87
20.850
14.084
−5.323
1.00
48.15
A


ATOM
627
C
THR
A
87
21.308
12.094
−8.581
1.00
48.08
A


ATOM
628
O
THR
A
87
20.289
12.120
−9.266
1.00
48.96
A


ATOM
629
N
GLY
A
88
22.515
11.868
−9.088
1.00
47.81
A


ATOM
630
CA
GLY
A
88
22.701
11.692
−10.520
1.00
47.20
A


ATOM
631
C
GLY
A
88
22.574
13.042
−11.212
1.00
47.00
A


ATOM
632
O
GLY
A
88
22.365
13.131
−12.424
1.00
47.22
A


ATOM
633
N
HIS
A
89
22.720
14.099
−10.419
1.00
45.03
A


ATOM
634
CA
HIS
A
89
22.601
15.482
−10.877
1.00
44.02
A


ATOM
635
CB
HIS
A
89
21.499
16.149
−10.033
1.00
45.14
A


ATOM
636
CG
HIS
A
89
21.222
17.577
−10.380
1.00
47.25
A


ATOM
637
CD2
HIS
A
89
20.207
18.143
−11.075
1.00
47.80
A


ATOM
638
ND1
HIS
A
89
22.043
18.612
−9.987
1.00
46.81
A


ATOM
639
CE1
HIS
A
89
21.545
19.755
−10.426
1.00
46.54
A


ATOM
640
NE2
HIS
A
89
20.432
19.499
−11.089
1.00
46.38
A


ATOM
641
C
HIS
A
89
23.962
16.183
−10.701
1.00
42.11
A


ATOM
642
O
HIS
A
89
24.802
15.715
−9.932
1.00
41.05
A


ATOM
643
N
PRO
A
90
24.208
17.292
−11.425
1.00
40.53
A


ATOM
644
CD
PRO
A
90
23.425
17.878
−12.529
1.00
41.84
A


ATOM
645
CA
PRO
A
90
25.492
17.990
−11.285
1.00
39.95
A


ATOM
646
CB
PRO
A
90
25.282
19.254
−12.103
1.00
39.62
A


ATOM
647
CG
PRO
A
90
24.447
18.759
−13.224
1.00
42.14
A


ATOM
648
C
PRO
A
90
25.879
18.295
−9.836
1.00
38.75
A


ATOM
649
O
PRO
A
90
27.062
18.400
−9.508
1.00
36.43
A


ATOM
650
N
VAL
A
91
24.884
18.432
−8.967
1.00
37.40
A


ATOM
651
CA
VAL
A
91
25.174
18.729
−7.573
1.00
35.42
A


ATOM
652
CB
VAL
A
91
23.863
18.890
−6.751
1.00
32.89
A


ATOM
653
CG1
VAL
A
91
23.089
17.585
−6.708
1.00
29.58
A


ATOM
654
CG2
VAL
A
91
24.188
19.390
−5.348
1.00
33.88
A


ATOM
655
C
VAL
A
91
26.070
17.647
−6.973
1.00
36.43
A


ATOM
656
O
VAL
A
91
26.809
17.897
−6.014
1.00
35.56
A


ATOM
657
N
ASP
A
92
26.041
16.453
−7.562
1.00
36.81
A


ATOM
658
CA
ASP
A
92
26.862
15.352
−7.064
1.00
37.47
A


ATOM
659
CB
ASP
A
92
26.513
14.033
−7.764
1.00
39.17
A


ATOM
660
CG
ASP
A
92
25.111
13.547
−7.441
1.00
42.34
A


ATOM
661
OD1
ASP
A
92
24.703
13.605
−6.257
1.00
42.11
A


ATOM
662
OD2
ASP
A
92
24.426
13.094
−8.379
1.00
41.11
A


ATOM
663
C
ASP
A
92
28.358
15.600
−7.241
1.00
37.40
A


ATOM
664
O
ASP
A
92
29.176
15.030
−6.522
1.00
36.00
A


ATOM
665
N
ASP
A
93
28.714
16.443
−8.200
1.00
37.21
A


ATOM
666
CA
ASP
A
93
30.119
16.714
−8.465
1.00
38.02
A


ATOM
667
CB
ASP
A
93
30.378
16.662
−9.974
1.00
39.00
A


ATOM
668
CG
ASP
A
93
29.861
15.385
−10.611
1.00
42.85
A


ATOM
669
OD1
ASP
A
93
30.238
14.293
−10.139
1.00
43.31
A


ATOM
670
OD2
ASP
A
93
29.075
15.476
−11.579
1.00
45.48
A


ATOM
671
C
ASP
A
93
30.625
18.044
−7.932
1.00
37.00
A


ATOM
672
O
ASP
A
93
31.831
18.245
−7.835
1.00
36.56
A


ATOM
673
N
LEU
A
94
29.710
18.938
−7.568
1.00
35.71
A


ATOM
674
CA
LEU
A
94
30.104
20.268
−7.120
1.00
34.14
A


ATOM
675
CB
LEU
A
94
28.867
21.131
−6.853
1.00
34.32
A


ATOM
676
CG
LEU
A
94
29.151
22.629
−6.678
1.00
33.22
A


ATOM
677
CD1
LEU
A
94
30.004
23.160
−7.823
1.00
33.67
A


ATOM
678
CD2
LEU
A
94
27.839
23.368
−6.627
1.00
34.83
A


ATOM
679
C
LEU
A
94
31.054
20.357
−5.931
1.00
33.54
A


ATOM
680
O
LEU
A
94
32.010
21.132
−5.971
1.00
32.88
A


ATOM
681
N
LEU
A
95
30.814
19.592
−4.874
1.00
33.38
A


ATOM
682
CA
LEU
A
95
31.729
19.687
−3.749
1.00
34.21
A


ATOM
683
CB
LEU
A
95
31.309
18.780
−2.589
1.00
31.65
A


ATOM
684
CG
LEU
A
95
32.185
18.988
−1.337
1.00
31.56
A


ATOM
685
CD1
LEU
A
95
32.251
20.480
−0.967
1.00
32.11
A


ATOM
686
CD2
LEU
A
95
31.620
18.187
−0.173
1.00
32.42
A


ATOM
687
C
LEU
A
95
33.111
19.300
−4.249
1.00
34.32
A


ATOM
688
O
LEU
A
95
34.074
20.036
−4.047
1.00
33.01
A


ATOM
689
N
ALA
A
96
33.197
18.158
−4.933
1.00
34.56
A


ATOM
690
CA
ALA
A
96
34.473
17.675
−5.457
1.00
35.45
A


ATOM
691
CB
ALA
A
96
34.284
16.303
−6.104
1.00
37.29
A


ATOM
692
C
ALA
A
96
35.132
18.641
−6.447
1.00
34.95
A


ATOM
693
O
ALA
A
96
36.340
18.899
−6.353
1.00
35.01
A


ATOM
694
N
ASP
A
97
34.358
19.175
−7.392
1.00
34.87
A


ATOM
695
CA
ASP
A
97
34.915
20.110
−8.377
1.00
34.02
A


ATOM
696
CB
ASP
A
97
33.874
20.486
−9.439
1.00
35.17
A


ATOM
697
CG
ASP
A
97
33.636
19.385
−10.457
1.00
36.22
A


ATOM
698
OD1
ASP
A
97
34.319
18.338
−10.405
1.00
37.31
A


ATOM
699
OD2
ASP
A
97
32.752
19.580
−11.318
1.00
35.57
A


ATOM
700
C
ASP
A
97
35.412
21.393
−7.725
1.00
34.14
A


ATOM
701
O
ASP
A
97
36.325
22.052
−8.232
1.00
33.33
A


ATOM
702
N
THR
A
98
34.791
21.757
−6.609
1.00
34.23
A


ATOM
703
CA
THR
A
98
35.155
22.966
−5.885
1.00
34.54
A


ATOM
704
CB
THR
A
98
34.095
23.286
−4.795
1.00
33.81
A


ATOM
705
OG1
THR
A
98
32.868
23.653
−5.433
1.00
32.38
A


ATOM
706
CG2
THR
A
98
34.551
24.430
−3.889
1.00
32.67
A


ATOM
707
C
THR
A
98
36.528
22.809
−5.242
1.00
35.23
A


ATOM
708
O
THR
A
98
37.356
23.722
−5.282
1.00
36.85
A


ATOM
709
N
GLN
A
99
36.764
21.645
−4.652
1.00
36.14
A


ATOM
710
CA
GLN
A
99
38.041
21.361
−4.005
1.00
38.58
A


ATOM
711
CB
GLN
A
99
37.919
20.077
−3.174
1.00
40.12
A


ATOM
712
CG
GLN
A
99
39.212
19.594
−2.530
1.00
44.11
A


ATOM
713
CD
GLN
A
99
38.962
18.628
−1.378
1.00
45.38
A


ATOM
714
OE1
GLN
A
99
38.046
17.800
−1.428
1.00
44.91
A


ATOM
715
NE2
GLN
A
99
39.784
18.723
−0.336
1.00
45.37
A


ATOM
716
C
GLN
A
99
39.121
21.224
−5.079
1.00
39.44
A


ATOM
717
O
GLN
A
99
40.309
21.422
−4.819
1.00
39.54
A


ATOM
718
N
LYS
A
100
38.693
20.899
−6.294
1.00
39.83
A


ATOM
719
CA
LYS
A
100
39.603
20.754
−7.423
1.00
41.72
A


ATOM
720
CB
LYS
A
100
38.913
19.961
−8.541
1.00
43.31
A


ATOM
721
CG
LYS
A
100
39.711
19.870
−9.841
1.00
47.29
A


ATOM
722
CD
LYS
A
100
38.870
19.299
−10.986
1.00
49.04
A


ATOM
723
CE
LYS
A
100
39.606
19.396
−12.327
1.00
50.86
A


ATOM
724
NZ
LYS
A
100
38.770
18.967
−13.495
1.00
52.06
A


ATOM
725
C
LYS
A
100
40.085
22.113
−7.970
1.00
41.64
A


ATOM
726
O
LYS
A
100
41.277
22.294
−8.241
1.00
42.73
A


ATOM
727
N
HIS
A
101
39.163
23.062
−8.126
1.00
40.66
A


ATOM
728
CA
HIS
A
101
39.494
24.378
−8.667
1.00
40.67
A


ATOM
729
CB
HIS
A
101
38.349
24.901
−9.546
1.00
41.87
A


ATOM
730
CG
HIS
A
101
38.153
24.138
−10.817
1.00
41.74
A


ATOM
731
CD2
HIS
A
101
38.455
24.453
−12.100
1.00
42.24
A


ATOM
732
ND1
HIS
A
101
37.587
22.882
−10.854
1.00
43.06
A


ATOM
733
CE1
HIS
A
101
37.547
22.456
−12.104
1.00
42.69
A


ATOM
734
NE2
HIS
A
101
38.069
23.390
−12.880
1.00
42.38
A


ATOM
735
C
HIS
A
101
39.835
25.475
−7.664
1.00
40.29
A


ATOM
736
O
HIS
A
101
40.307
26.537
−8.065
1.00
41.90
A


ATOM
737
N
LEU
A
102
39.598
25.243
−6.376
1.00
39.72
A


ATOM
738
CA
LEU
A
102
39.869
26.279
−5.378
1.00
38.61
A


ATOM
739
CB
LEU
A
102
38.589
27.073
−5.068
1.00
37.62
A


ATOM
740
CG
LEU
A
102
37.875
27.877
−6.156
1.00
37.33
A


ATOM
741
CD1
LEU
A
102
36.523
28.356
−5.621
1.00
37.68
A


ATOM
742
CD2
LEU
A
102
38.736
29.062
−6.586
1.00
38.82
A


ATOM
743
C
LEU
A
102
40.412
25.735
−4.069
1.00
37.71
A


ATOM
744
O
LEU
A
102
40.259
24.558
−3.756
1.00
37.42
A


ATOM
745
N
PRO
A
103
41.055
26.603
−3.279
1.00
38.28
A


ATOM
746
CD
PRO
A
103
41.519
27.947
−3.660
1.00
38.45
A


ATOM
747
CA
PRO
A
103
41.620
26.209
−1.988
1.00
38.75
A


ATOM
748
CB
PRO
A
103
42.559
27.367
−1.647
1.00
39.03
A


ATOM
749
CG
PRO
A
103
42.849
28.006
−2.977
1.00
39.90
A


ATOM
750
C
PRO
A
103
40.501
26.084
−0.962
1.00
38.05
A


ATOM
751
O
PRO
A
103
39.871
27.079
−0.608
1.00
39.88
A


ATOM
752
N
VAL
A
104
40.232
24.867
−0.507
1.00
38.36
A


ATOM
753
CA
VAL
A
104
39.196
24.658
0.492
1.00
38.23
A


ATOM
754
CB
VAL
A
104
38.300
23.457
0.126
1.00
37.32
A


ATOM
755
CG1
VAL
A
104
37.288
23.195
1.226
1.00
35.34
A


ATOM
756
CG2
VAL
A
104
37.582
23.748
−1.190
1.00
37.51
A


ATOM
757
C
VAL
A
104
39.902
24.426
1.819
1.00
39.33
A


ATOM
758
O
VAL
A
104
40.699
23.503
1.963
1.00
39.67
A


ATOM
759
N
SER
A
105
39.604
25.284
2.783
1.00
39.48
A


ATOM
760
CA
SER
A
105
40.228
25.230
4.098
1.00
38.55
A


ATOM
761
CB
SER
A
105
40.410
26.658
4.613
1.00
37.61
A


ATOM
762
OG
SER
A
105
39.157
27.324
4.641
1.00
38.92
A


ATOM
763
C
SER
A
105
39.452
24.421
5.127
1.00
37.52
A


ATOM
764
O
SER
A
105
39.991
24.060
6.176
1.00
36.61
A


ATOM
765
N
MET
A
106
38.188
24.132
4.836
1.00
36.90
A


ATOM
766
CA
MET
A
106
37.368
23.387
5.773
1.00
35.42
A


ATOM
767
CB
MET
A
106
37.114
24.245
7.013
1.00
39.45
A


ATOM
768
CG
MET
A
106
36.289
23.574
8.088
1.00
44.17
A


ATOM
769
SD
MET
A
106
36.685
24.199
9.727
1.00
52.21
A


ATOM
770
CE
MET
A
106
35.296
25.277
10.026
1.00
49.49
A


ATOM
771
C
MET
A
106
36.036
23.000
5.147
1.00
34.14
A


ATOM
772
O
MET
A
106
35.596
23.624
4.187
1.00
31.92
A


ATOM
773
N
PHE
A
107
35.413
21.969
5.706
1.00
31.68
A


ATOM
774
CA
PHE
A
107
34.116
21.479
5.251
1.00
32.11
A


ATOM
775
CB
PHE
A
107
34.186
19.993
4.869
1.00
32.56
A


ATOM
776
CG
PHE
A
107
35.046
19.685
3.674
1.00
32.69
A


ATOM
777
CD1
PHE
A
107
36.107
18.789
3.787
1.00
33.01
A


ATOM
778
CD2
PHE
A
107
34.756
20.223
2.425
1.00
34.08
A


ATOM
779
CE1
PHE
A
107
36.865
18.429
2.675
1.00
33.52
A


ATOM
780
CE2
PHE
A
107
35.514
19.867
1.296
1.00
35.10
A


ATOM
781
CZ
PHE
A
107
36.565
18.969
1.426
1.00
34.67
A


ATOM
782
C
PHE
A
107
33.107
21.581
6.395
1.00
30.99
A


ATOM
783
O
PHE
A
107
33.445
21.943
7.525
1.00
30.91
A


ATOM
784
N
ALA
A
108
31.861
21.248
6.082
1.00
31.37
A


ATOM
785
CA
ALA
A
108
30.798
21.204
7.071
1.00
29.87
A


ATOM
786
CB
ALA
A
108
30.384
22.603
7.494
1.00
29.38
A


ATOM
787
C
ALA
A
108
29.629
20.457
6.441
1.00
30.97
A


ATOM
788
O
ALA
A
108
29.518
20.383
5.211
1.00
29.63
A


ATOM
789
N
ILE
A
109
28.790
19.855
7.274
1.00
31.61
A


ATOM
790
CA
ILE
A
109
27.624
19.156
6.763
1.00
33.76
A


ATOM
791
CB
ILE
A
109
27.718
17.627
6.925
1.00
35.00
A


ATOM
792
CG2
ILE
A
109
28.841
17.086
6.047
1.00
34.55
A


ATOM
793
CG1
ILE
A
109
27.911
17.261
8.398
1.00
35.82
A


ATOM
794
CD1
ILE
A
109
27.901
15.782
8.661
1.00
39.05
A


ATOM
795
C
ILE
A
109
26.427
19.671
7.524
1.00
35.47
A


ATOM
796
O
ILE
A
109
26.553
20.197
8.634
1.00
34.00
A


ATOM
797
N
ASP
A
110
25.262
19.526
6.911
1.00
35.28
A


ATOM
798
CA
ASP
A
110
24.032
20.002
7.499
1.00
36.47
A


ATOM
799
CB
ASP
A
110
23.577
21.231
6.710
1.00
39.55
A


ATOM
800
CG
ASP
A
110
23.876
22.526
7.408
1.00
42.35
A


ATOM
801
OD1
ASP
A
110
24.816
22.555
8.227
1.00
42.34
A


ATOM
802
OD2
ASP
A
110
23.155
23.514
7.144
1.00
45.22
A


ATOM
803
C
ASP
A
110
22.997
18.886
7.419
1.00
34.76
A


ATOM
804
O
ASP
A
110
22.944
18.147
6.437
1.00
31.82
A


ATOM
805
N
GLY
A
111
22.195
18.762
8.470
1.00
33.94
A


ATOM
806
CA
GLY
A
111
21.156
17.755
8.509
1.00
35.79
A


ATOM
807
C
GLY
A
111
19.931
18.330
9.201
1.00
35.96
A


ATOM
808
O
GLY
A
111
19.957
19.465
9.672
1.00
35.30
A


ATOM
809
N
GLU
A
112
18.855
17.553
9.244
1.00
37.97
A


ATOM
810
CA
GLU
A
112
17.613
17.962
9.893
1.00
38.66
A


ATOM
811
CB
GLU
A
112
16.502
18.123
8.850
1.00
39.80
A


ATOM
812
CG
GLU
A
112
15.336
19.019
9.264
1.00
38.95
A


ATOM
813
CD
GLU
A
112
14.378
18.352
10.240
1.00
40.97
A


ATOM
814
OE1
GLU
A
112
13.934
17.222
9.952
1.00
41.18
A


ATOM
815
OE2
GLU
A
112
14.060
18.964
11.285
1.00
40.91
A


ATOM
816
C
GLU
A
112
17.311
16.810
10.845
1.00
39.79
A


ATOM
817
O
GLU
A
112
17.540
15.654
10.505
1.00
40.11
A


ATOM
818
N
VAL
A
113
16.808
17.118
12.034
1.00
41.22
A


ATOM
819
CA
VAL
A
113
16.536
16.082
13.027
1.00
42.17
A


ATOM
820
CB
VAL
A
113
16.051
16.700
14.363
1.00
41.43
A


ATOM
821
CG1
VAL
A
113
17.131
17.595
14.931
1.00
40.29
A


ATOM
822
CG2
VAL
A
113
14.762
17.478
14.153
1.00
39.96
A


ATOM
823
C
VAL
A
113
15.557
14.989
12.616
1.00
43.71
A


ATOM
824
O
VAL
A
113
15.492
13.939
13.251
1.00
45.17
A


ATOM
825
N
THR
A
114
14.802
15.222
11.553
1.00
45.21
A


ATOM
826
CA
THR
A
114
13.843
14.226
11.104
1.00
46.88
A


ATOM
827
CB
THR
A
114
12.410
14.782
11.146
1.00
48.04
A


ATOM
828
CG1
THR
A
114
12.160
15.360
12.433
1.00
51.16
A


ATOM
829
CG2
THR
A
114
11.405
13.675
10.900
1.00
50.04
A


ATOM
830
C
THR
A
114
14.126
13.764
9.681
1.00
46.51
A


ATOM
831
O
THR
A
114
13.642
12.715
9.256
1.00
47.78
A


ATOM
832
N
GLY
A
115
14.910
14.545
8.946
1.00
43.52
A


ATOM
833
CA
GLY
A
115
15.202
14.182
7.572
1.00
43.24
A


ATOM
834
C
GLY
A
115
16.606
13.680
7.324
1.00
41.33
A


ATOM
835
O
GLY
A
115
16.873
13.058
6.294
1.00
42.99
A


ATOM
836
N
GLY
A
116
17.507
13.954
8.259
1.00
40.18
A


ATOM
837
CA
GLY
A
116
18.881
13.509
8.102
1.00
38.86
A


ATOM
838
C
GLY
A
116
19.699
14.416
7.202
1.00
36.69
A


ATOM
839
O
GLY
A
116
19.308
15.543
6.940
1.00
36.36
A


ATOM
840
N
PHE
A
117
20.827
13.905
6.724
1.00
36.25
A


ATOM
841
CA
PHE
A
117
21.749
14.649
5.862
1.00
35.53
A


ATOM
842
CB
PHE
A
117
22.828
13.706
5.325
1.00
34.63
A


ATOM
843
CG
PHE
A
117
23.756
14.350
4.329
1.00
33.58
A


ATOM
844
CD1
PHE
A
117
24.874
15.059
4.755
1.00
32.41
A


ATOM
845
CD2
PHE
A
117
23.502
14.255
2.961
1.00
32.56
A


ATOM
846
CE1
PHE
A
117
25.737
15.666
3.836
1.00
31.07
A


ATOM
847
CE2
PHE
A
117
24.357
14.859
2.032
1.00
31.84
A


ATOM
848
CZ
PHE
A
117
25.479
15.566
2.474
1.00
31.62
A


ATOM
849
C
PHE
A
117
21.081
15.337
4.675
1.00
35.92
A


ATOM
850
O
PHE
A
117
20.229
14.743
4.012
1.00
35.06
A


ATOM
851
N
LYS
A
118
21.491
16.576
4.395
1.00
34.72
A


ATOM
852
CA
LYS
A
118
20.937
17.315
3.265
1.00
34.75
A


ATOM
853
CB
LYS
A
118
19.691
18.098
3.698
1.00
36.47
A


ATOM
854
CG
LYS
A
118
19.918
19.142
4.783
1.00
37.72
A


ATOM
855
CD
LYS
A
118
18.714
20.079
4.867
1.00
40.72
A


ATOM
856
CE
LYS
A
118
18.968
21.253
5.807
1.00
39.89
A


ATOM
857
NZ
LYS
A
118
17.892
22.284
5.712
1.00
39.12
A


ATOM
858
C
LYS
A
118
21.920
18.251
2.546
1.00
32.27
A


ATOM
859
O
LYS
A
118
21.713
18.584
1.380
1.00
29.66
A


ATOM
860
N
LYS
A
119
22.992
18.672
3.218
1.00
31.59
A


ATOM
861
CA
LYS
A
119
23.955
19.551
2.564
1.00
30.61
A


ATOM
862
CB
LYS
A
119
23.400
20.985
2.489
1.00
34.11
A


ATOM
863
CG
LYS
A
119
22.923
21.579
3.783
1.00
34.64
A


ATOM
864
CD
LYS
A
119
22.230
22.956
3.600
1.00
33.28
A


ATOM
865
CE
LYS
A
119
23.155
23.995
2.982
1.00
32.16
A


ATOM
866
NZ
LYS
A
119
22.817
25.404
3.328
1.00
29.97
A


ATOM
867
C
LYS
A
119
25.381
19.578
3.110
1.00
30.58
A


ATOM
868
O
LYS
A
119
25.657
19.174
4.246
1.00
28.47
A


ATOM
869
N
THR
A
120
26.298
20.021
2.258
1.00
31.56
A


ATOM
870
CA
THR
A
120
27.702
20.156
2.637
1.00
30.27
A


ATOM
871
CB
THR
A
120
28.645
19.319
1.749
1.00
29.70
A


ATOM
872
OG1
THR
A
120
28.467
19.710
0.381
1.00
28.77
A


ATOM
873
CG2
THR
A
120
28.370
17.841
1.904
1.00
28.77
A


ATOM
874
C
THR
A
120
28.071
21.605
2.417
1.00
30.38
A


ATOM
875
O
THR
A
120
27.333
22.356
1.760
1.00
29.15
A


ATOM
876
N
TYR
A
121
29.226
21.984
2.952
1.00
28.30
A


ATOM
877
CA
TYR
A
121
29.740
23.332
2.817
1.00
29.10
A


ATOM
878
CB
TYR
A
121
29.632
24.107
4.136
1.00
30.57
A


ATOM
879
CG
TYR
A
121
28.233
24.402
4.611
1.00
31.61
A


ATOM
880
CD1
TYR
A
121
27.450
23.416
5.208
1.00
32.95
A


ATOM
881
CE1
TYR
A
121
26.160
23.692
5.650
1.00
33.00
A


ATOM
882
CD2
TYR
A
121
27.692
25.674
4.468
1.00
33.43
A


ATOM
883
CE2
TYR
A
121
26.406
25.959
4.903
1.00
35.44
A


ATOM
884
CZ
TYR
A
121
25.648
24.966
5.488
1.00
33.78
A


ATOM
885
OH
TYR
A
121
24.369
25.259
5.885
1.00
36.41
A


ATOM
886
C
TYR
A
121
31.215
23.250
2.461
1.00
28.23
A


ATOM
887
O
TYR
A
121
31.916
22.330
2.881
1.00
27.61
A


ATOM
888
N
ALA
A
122
31.675
24.212
1.679
1.00
29.43
A


ATOM
889
CA
ALA
A
122
33.079
24.284
1.324
1.00
30.31
A


ATOM
890
CB
ALA
A
122
33.258
24.126
−0.168
1.00
29.87
A


ATOM
891
C
ALA
A
122
33.512
25.677
1.775
1.00
30.35
A


ATOM
892
O
ALA
A
122
32.971
26.675
1.305
1.00
30.95
A


ATOM
893
N
PHE
A
123
34.456
25.740
2.710
1.00
30.19
A


ATOM
894
CA
PHE
A
123
34.945
27.023
3.212
1.00
31.04
A


ATOM
895
CB
PHE
A
123
35.185
26.982
4.728
1.00
32.77
A


ATOM
896
CG
PHE
A
123
33.937
26.889
5.543
1.00
33.65
A


ATOM
897
CD1
PHE
A
123
33.215
25.702
5.601
1.00
34.51
A


ATOM
898
CD2
PHE
A
123
33.489
27.988
6.273
1.00
36.59
A


ATOM
899
CE1
PHE
A
123
32.054
25.608
6.384
1.00
36.52
A


ATOM
900
CE2
PHE
A
123
32.330
27.908
7.061
1.00
36.10
A


ATOM
901
CZ
PHE
A
123
31.616
26.719
7.115
1.00
34.21
A


ATOM
902
C
PHE
A
123
36.257
27.385
2.547
1.00
32.63
A


ATOM
903
O
PHE
A
123
37.079
26.507
2.265
1.00
34.75
A


ATOM
904
N
PHE
A
124
36.467
28.676
2.312
1.00
32.86
A


ATOM
905
CA
PHE
A
124
37.706
29.123
1.702
1.00
33.91
A


ATOM
906
CB
PHE
A
124
37.410
29.930
0.441
1.00
33.32
A


ATOM
907
CG
PHE
A
124
36.423
29.261
−0.479
1.00
32.63
A


ATOM
908
CD1
PHE
A
124
35.106
29.712
−0.555
1.00
31.58
A


ATOM
909
CD2
PHE
A
124
36.797
28.158
−1.241
1.00
33.35
A


ATOM
910
CE1
PHE
A
124
34.179
29.068
−1.377
1.00
34.49
A


ATOM
911
CE2
PHE
A
124
35.877
27.504
−2.067
1.00
33.34
A


ATOM
912
CZ
PHE
A
124
34.569
27.957
−2.135
1.00
33.17
A


ATOM
913
C
PHE
A
124
38.495
29.959
2.703
1.00
33.67
A


ATOM
914
O
PHE
A
124
37.940
30.463
3.681
1.00
33.97
A


ATOM
915
N
PRO
A
125
39.814
30.079
2.495
1.00
35.21
A


ATOM
916
CD
PRO
A
125
40.642
29.338
1.527
1.00
35.76
A


ATOM
917
CA
PRO
A
125
40.660
30.866
3.397
1.00
34.83
A


ATOM
918
CB
PRO
A
125
42.048
30.718
2.776
1.00
35.56
A


ATOM
919
CG
PRO
A
125
41.995
29.336
2.207
1.00
35.73
A


ATOM
920
C
PRO
A
125
40.192
32.310
3.439
1.00
34.10
A


ATOM
921
O
PRO
A
125
39.948
32.925
2.405
1.00
34.97
A


ATOM
922
N
THR
A
126
40.056
32.846
4.645
1.00
36.61
A


ATOM
923
CA
THR
A
126
39.605
34.217
4.822
1.00
38.10
A


ATOM
924
CB
THR
A
126
39.556
34.572
6.319
1.00
37.99
A


ATOM
925
OG1
THR
A
126
38.725
33.620
6.991
1.00
42.36
A


ATOM
926
CG2
THR
A
126
38.977
35.966
6.535
1.00
41.12
A


ATOM
927
C
THR
A
126
40.492
35.218
4.085
1.00
37.53
A


ATOM
928
O
THR
A
126
40.014
36.258
3.635
1.00
37.06
A


ATOM
929
N
ASP
A
127
41.777
34.897
3.940
1.00
39.58
A


ATOM
930
CA
ASP
A
127
42.701
35.801
3.263
1.00
39.86
A


ATOM
931
CB
ASP
A
127
44.026
35.896
4.034
1.00
42.23
A


ATOM
932
CG
ASP
A
127
44.763
34.567
4.124
1.00
43.17
A


ATOM
933
OD1
ASP
A
127
44.243
33.539
3.641
1.00
45.25
A


ATOM
934
OD2
ASP
A
127
45.878
34.554
4.688
1.00
45.84
A


ATOM
935
C
ASP
A
127
42.973
35.435
1.810
1.00
39.84
A


ATOM
936
O
ASP
A
127
43.943
35.904
1.221
1.00
40.14
A


ATOM
937
N
ASN
A
128
42.109
34.600
1.238
1.00
39.79
A


ATOM
938
CA
ASN
A
128
42.238
34.182
−0.153
1.00
39.11
A


ATOM
939
CB
ASN
A
128
43.332
33.121
−0.305
1.00
40.96
A


ATOM
940
CG
ASN
A
128
43.608
32.772
−1.763
1.00
42.76
A


ATOM
941
OD1
ASN
A
128
44.327
31.816
−2.058
1.00
45.02
A


ATOM
942
ND2
ASN
A
128
43.044
33.548
−2.679
1.00
43.31
A


ATOM
943
C
ASN
A
128
40.910
33.600
−0.611
1.00
38.19
A


ATOM
944
O
ASN
A
128
40.834
32.444
−1.027
1.00
36.34
A


ATOM
945
N
MET
A
129
39.860
34.407
−0.532
1.00
37.03
A


ATOM
946
CA
MET
A
129
38.544
33.948
−0.933
1.00
36.26
A


ATOM
947
CB
MET
A
129
37.462
34.742
−0.205
1.00
35.19
A


ATOM
948
CG
MET
A
129
37.469
34.578
1.300
1.00
36.19
A


ATOM
949
SD
MET
A
129
36.104
35.498
2.022
1.00
35.06
A


ATOM
950
CE
MET
A
129
36.869
37.133
2.125
1.00
37.35
A


ATOM
951
C
MET
A
129
38.375
34.127
−2.431
1.00
35.46
A


ATOM
952
O
MET
A
129
38.953
35.036
−3.024
1.00
35.81
A


ATOM
953
N
PRO
A
130
37.587
33.252
−3.067
1.00
35.15
A


ATOM
954
CD
PRO
A
130
37.024
31.988
−2.558
1.00
33.45
A


ATOM
955
CA
PRO
A
130
37.375
33.374
−4.510
1.00
34.52
A


ATOM
956
CB
PRO
A
130
36.935
31.968
−4.907
1.00
34.77
A


ATOM
957
CG
PRO
A
130
36.155
31.528
−3.713
1.00
33.09
A


ATOM
958
C
PRO
A
130
36.313
34.428
−4.825
1.00
35.76
A


ATOM
959
O
PRO
A
130
35.502
34.785
−3.966
1.00
34.18
A


ATOM
960
N
GLY
A
131
36.343
34.936
−6.055
1.00
36.06
A


ATOM
961
CA
GLY
A
131
35.371
35.921
−6.483
1.00
36.41
A


ATOM
962
C
GLY
A
131
34.313
35.190
−7.282
1.00
37.83
A


ATOM
963
O
GLY
A
131
34.424
33.981
−7.482
1.00
38.41
A


ATOM
964
N
VAL
A
132
33.288
35.900
−7.739
1.00
37.75
A


ATOM
965
CA
VAL
A
132
32.229
35.269
−8.515
1.00
39.80
A


ATOM
966
CB
VAL
A
132
31.106
36.277
−8.848
1.00
39.90
A


ATOM
967
CG1
VAL
A
132
30.089
35.649
−9.792
1.00
39.29
A


ATOM
968
CG2
VAL
A
132
30.420
36.711
−7.569
1.00
39.79
A


ATOM
969
C
VAL
A
132
32.795
34.688
−9.804
1.00
40.83
A


ATOM
970
O
VAL
A
132
32.388
33.615
−10.245
1.00
41.10
A


ATOM
971
N
ALA
A
133
33.747
35.397
−10.398
1.00
42.28
A


ATOM
972
CA
ALA
A
133
34.378
34.949
−11.632
1.00
43.17
A


ATOM
973
CB
ALA
A
133
35.482
35.923
−12.036
1.00
44.08
A


ATOM
974
C
ALA
A
133
34.955
33.548
−11.462
1.00
43.96
A


ATOM
975
O
ALA
A
133
34.636
32.651
−12.235
1.00
43.99
A


ATOM
976
N
GLU
A
134
35.799
33.366
−10.446
1.00
44.05
A


ATOM
977
CA
GLU
A
134
36.421
32.067
−10.173
1.00
44.20
A


ATOM
978
CB
GLU
A
134
37.366
32.159
−8.969
1.00
45.02
A


ATOM
979
CG
GLU
A
134
38.507
33.150
−9.119
1.00
48.35
A


ATOM
980
CD
GLU
A
134
38.168
34.527
−8.579
1.00
50.23
A


ATOM
981
OE1
GLU
A
134
37.190
35.148
−9.057
1.00
51.33
A


ATOM
982
OE2
GLU
A
134
38.889
34.991
−7.670
1.00
52.57
A


ATOM
983
C
GLU
A
134
35.406
30.954
−9.905
1.00
42.84
A


ATOM
984
O
GLU
A
134
35.587
29.821
−10.348
1.00
44.08
A


ATOM
985
N
LEU
A
135
34.343
31.272
−9.175
1.00
41.08
A


ATOM
986
CA
LEU
A
135
33.325
30.276
−8.859
1.00
40.14
A


ATOM
987
CB
LEU
A
135
32.349
30.827
−7.808
1.00
38.00
A


ATOM
988
CG
LEU
A
135
32.858
30.961
−6.363
1.00
35.55
A


ATOM
989
CD1
LEU
A
135
31.813
31.634
−5.501
1.00
37.80
A


ATOM
990
CD2
LEU
A
135
33.180
29.589
−5.807
1.00
32.97
A


ATOM
991
C
LEU
A
135
32.544
29.786
−10.079
1.00
41.44
A


ATOM
992
O
LEU
A
135
32.419
28.583
−10.294
1.00
41.03
A


ATOM
993
N
SER
A
136
32.018
30.710
−10.877
1.00
43.65
A


ATOM
994
CA
SER
A
136
31.234
30.341
−12.064
1.00
45.88
A


ATOM
995
CB
SER
A
136
30.756
31.599
−12.798
1.00
47.35
A


ATOM
996
OG
SER
A
136
31.851
32.390
−13.227
1.00
49.44
A


ATOM
997
C
SER
A
136
31.995
29.448
−13.041
1.00
45.67
A


ATOM
998
O
SER
A
136
31.395
28.799
−13.903
1.00
46.58
A


ATOM
999
N
ALA
A
137
33.316
29.416
−12.892
1.00
45.23
A


ATOM
1000
CA
ALA
A
137
34.185
28.618
−13.746
1.00
44.09
A


ATOM
1001
CB
ALA
A
137
35.618
29.123
−13.627
1.00
45.40
A


ATOM
1002
C
ALA
A
137
34.128
27.127
−13.413
1.00
43.23
A


ATOM
1003
O
ALA
A
137
34.433
26.284
−14.258
1.00
42.56
A


ATOM
1004
N
ILE
A
138
33.758
26.799
−12.179
1.00
41.63
A


ATOM
1005
CA
ILE
A
138
33.664
25.398
−11.775
1.00
40.86
A


ATOM
1006
CB
ILE
A
138
33.260
25.275
−10.280
1.00
39.46
A


ATOM
1007
CG2
ILE
A
138
33.120
23.812
−9.893
1.00
39.81
A


ATOM
1008
CG1
ILE
A
138
34.319
25.955
−9.403
1.00
40.37
A


ATOM
1009
CD1
ILE
A
138
33.990
25.985
−7.921
1.00
38.95
A


ATOM
1010
C
ILE
A
138
32.600
24.771
−12.677
1.00
40.15
A


ATOM
1011
O
ILE
A
138
31.481
25.261
−12.745
1.00
39.48
A


ATOM
1012
N
PRO
A
139
32.949
23.688
−13.391
1.00
42.04
A


ATOM
1013
CD
PRO
A
139
34.277
23.056
−13.319
1.00
42.63
A


ATOM
1014
CA
PRO
A
139
32.080
22.950
−14.325
1.00
42.17
A


ATOM
1015
CB
PRO
A
139
32.913
21.716
−14.668
1.00
43.33
A


ATOM
1016
CG
PRO
A
139
34.308
22.234
−14.591
1.00
44.53
A


ATOM
1017
C
PRO
A
139
30.672
22.568
−13.873
1.00
41.62
A


ATOM
1018
O
PRO
A
139
29.717
22.686
−14.646
1.00
41.11
A


ATOM
1019
N
SER
A
140
30.548
22.103
−12.634
1.00
40.71
A


ATOM
1020
CA
SER
A
140
29.262
21.677
−12.095
1.00
40.02
A


ATOM
1021
CB
SER
A
140
29.458
20.440
−11.207
1.00
38.55
A


ATOM
1022
OG
SER
A
140
30.473
20.655
−10.239
1.00
37.26
A


ATOM
1023
C
SER
A
140
28.510
22.765
−11.329
1.00
40.37
A


ATOM
1024
O
SER
A
140
27.460
22.507
−10.735
1.00
40.00
A


ATOM
1025
N
MET
A
141
29.044
23.981
−11.351
1.00
39.35
A


ATOM
1026
CA
MET
A
141
28.409
25.107
−10.671
1.00
38.79
A


ATOM
1027
CB
MET
A
141
29.383
26.292
−10.608
1.00
38.90
A


ATOM
1028
CG
MET
A
141
28.904
27.492
−9.790
1.00
39.10
A


ATOM
1029
SD
MET
A
141
28.779
27.156
−8.006
1.00
38.89
A


ATOM
1030
CE
MET
A
141
30.511
27.219
−7.524
1.00
39.82
A


ATOM
1031
C
MET
A
141
27.167
25.500
−11.474
1.00
38.43
A


ATOM
1032
O
MET
A
141
27.168
25.418
−12.704
1.00
38.34
A


ATOM
1033
N
PRO
A
142
26.088
25.924
−10.794
1.00
38.99
A


ATOM
1034
CD
PRO
A
142
25.876
26.042
−9.340
1.00
37.35
A


ATOM
1035
CA
PRO
A
142
24.882
26.314
−11.535
1.00
37.96
A


ATOM
1036
CB
PRO
A
142
23.949
26.823
−10.439
1.00
38.94
A


ATOM
1037
CG
PRO
A
142
24.372
26.043
−9.235
1.00
37.01
A


ATOM
1038
C
PRO
A
142
25.210
27.413
−12.545
1.00
39.39
A


ATOM
1039
O
PRO
A
142
25.983
28.329
−12.248
1.00
38.95
A


ATOM
1040
N
PRO
A
143
24.644
27.328
−13.758
1.00
37.81
A


ATOM
1041
CD
PRO
A
143
23.843
26.222
−14.307
1.00
39.42
A


ATOM
1042
CA
PRO
A
143
24.903
28.344
−14.780
1.00
38.06
A


ATOM
1043
CB
PRO
A
143
24.060
27.863
−15.964
1.00
38.66
A


ATOM
1044
CG
PRO
A
143
24.053
26.383
−15.793
1.00
39.51
A


ATOM
1045
C
PRO
A
143
24.439
29.708
−14.266
1.00
38.30
A


ATOM
1046
O
PRO
A
143
24.955
30.755
−14.665
1.00
37.08
A


ATOM
1047
N
ALA
A
144
23.462
29.670
−13.364
1.00
37.27
A


ATOM
1048
CA
ALA
A
144
22.887
30.871
−12.773
1.00
37.43
A


ATOM
1049
CB
ALA
A
144
21.813
30.485
−11.767
1.00
35.79
A


ATOM
1050
C
ALA
A
144
23.910
31.793
−12.115
1.00
36.34
A


ATOM
1051
O
ALA
A
144
23.700
33.002
−12.047
1.00
37.01
A


ATOM
1052
N
VAL
A
145
25.018
31.241
−11.636
1.00
35.52
A


ATOM
1053
CA
VAL
A
145
26.020
32.087
−10.993
1.00
36.56
A


ATOM
1054
CB
VAL
A
145
27.105
31.231
−10.292
1.00
33.87
A


ATOM
1055
CG1
VAL
A
145
28.206
32.118
−9.755
1.00
32.14
A


ATOM
1056
CG2
VAL
A
145
26.475
30.421
−9.164
1.00
33.34
A


ATOM
1057
C
VAL
A
145
26.674
33.047
−12.000
1.00
37.24
A


ATOM
1058
O
VAL
A
145
26.787
34.246
−11.746
1.00
35.40
A


ATOM
1059
N
ALA
A
146
27.093
32.524
−13.146
1.00
38.95
A


ATOM
1060
CA
ALA
A
146
27.713
33.365
−14.165
1.00
40.89
A


ATOM
1061
CB
ALA
A
146
28.233
32.505
−15.323
1.00
41.08
A


ATOM
1062
C
ALA
A
146
26.695
34.374
−14.679
1.00
41.46
A


ATOM
1063
O
ALA
A
146
27.013
35.543
−14.889
1.00
41.33
A


ATOM
1064
N
GLU
A
147
25.464
33.917
−14.866
1.00
42.76
A


ATOM
1065
CA
GLU
A
147
24.405
34.777
−15.367
1.00
44.04
A


ATOM
1066
CB
GLU
A
147
23.161
33.938
−15.657
1.00
46.32
A


ATOM
1067
CG
GLU
A
147
23.296
33.089
−16.924
1.00
51.66
A


ATOM
1068
CD
GLU
A
147
22.729
31.687
−16.774
1.00
53.70
A


ATOM
1069
OE1
GLU
A
147
21.566
31.555
−16.334
1.00
55.88
A


ATOM
1070
OE2
GLU
A
147
23.449
30.717
−17.104
1.00
56.34
A


ATOM
1071
C
GLU
A
147
24.066
35.923
−14.421
1.00
43.26
A


ATOM
1072
O
GLU
A
147
23.458
36.910
−14.833
1.00
41.77
A


ATOM
1073
N
ASN
A
148
24.474
35.797
−13.162
1.00
42.18
A


ATOM
1074
CA
ASN
A
148
24.200
36.827
−12.165
1.00
41.99
A


ATOM
1075
CB
ASN
A
148
23.588
36.193
−10.911
1.00
42.15
A


ATOM
1076
CG
ASN
A
148
22.104
35.918
−11.059
1.00
40.63
A


ATOM
1077
OD1
ASN
A
148
21.301
36.841
−11.145
1.00
42.33
A


ATOM
1078
ND2
ASN
A
148
21.735
34.649
−11.093
1.00
41.13
A


ATOM
1079
C
ASN
A
148
25.435
37.630
−11.774
1.00
41.70
A


ATOM
1080
O
ASN
A
148
25.361
38.492
−10.904
1.00
41.52
A


ATOM
1081
N
ALA
A
149
26.560
37.350
−12.427
1.00
42.05
A


ATOM
1082
CA
ALA
A
149
27.825
38.027
−12.150
1.00
42.82
A


ATOM
1083
CB
ALA
A
149
28.898
37.556
−13.143
1.00
41.85
A


ATOM
1084
C
ALA
A
149
27.743
39.553
−12.169
1.00
43.54
A


ATOM
1085
O
ALA
A
149
28.236
40.217
−11.257
1.00
44.06
A


ATOM
1086
N
GLU
A
150
27.132
40.103
−13.215
1.00
44.98
A


ATOM
1087
CA
GLU
A
150
26.997
41.549
−13.355
1.00
45.33
A


ATOM
1088
CB
GLU
A
150
26.373
41.891
−14.708
1.00
47.44
A


ATOM
1089
CG
GLU
A
150
27.297
41.639
−15.876
1.00
52.19
A


ATOM
1090
CD
GLU
A
150
26.605
41.834
−17.206
1.00
54.21
A


ATOM
1091
OE1
GLU
A
150
25.720
41.018
−17.540
1.00
56.56
A


ATOM
1092
OE2
GLU
A
150
26.936
42.808
−17.912
1.00
54.57
A


ATOM
1093
C
GLU
A
150
26.156
42.145
−12.241
1.00
43.75
A


ATOM
1094
O
GLU
A
150
26.485
43.200
−11.695
1.00
43.51
A


ATOM
1095
N
LEU
A
151
25.063
41.468
−11.915
1.00
42.63
A


ATOM
1096
CA
LEU
A
151
24.181
41.922
−10.853
1.00
41.40
A


ATOM
1097
CB
LEU
A
151
22.963
40.995
−10.777
1.00
42.33
A


ATOM
1098
CG
LEU
A
151
21.804
41.368
−9.850
1.00
42.54
A


ATOM
1099
CD1
LEU
A
151
20.538
40.668
−10.323
1.00
43.25
A


ATOM
1100
CD2
LEU
A
151
22.139
40.989
−8.414
1.00
41.89
A


ATOM
1101
C
LEU
A
151
24.954
41.941
−9.522
1.00
40.27
A


ATOM
1102
O
LEU
A
151
24.973
42.956
−8.822
1.00
41.45
A


ATOM
1103
N
PHE
A
152
25.597
40.826
−9.181
1.00
38.34
A


ATOM
1104
CA
PHE
A
152
26.372
40.745
−7.946
1.00
36.94
A


ATOM
1105
CB
PHE
A
152
27.129
39.417
−7.870
1.00
34.62
A


ATOM
1106
CG
PHE
A
152
26.249
38.217
−7.670
1.00
33.59
A


ATOM
1107
CD1
PHE
A
152
26.625
36.981
−8.183
1.00
32.62
A


ATOM
1108
CD2
PHE
A
152
25.071
38.307
−6.936
1.00
31.47
A


ATOM
1109
CE1
PHE
A
152
25.842
35.844
−7.969
1.00
32.66
A


ATOM
1110
CE2
PHE
A
152
24.281
37.177
−6.714
1.00
31.36
A


ATOM
1111
CZ
PHE
A
152
24.670
35.941
−7.232
1.00
32.67
A


ATOM
1112
C
PHE
A
152
27.379
41.891
−7.889
1.00
38.16
A


ATOM
1113
O
PHE
A
152
27.499
42.575
−6.874
1.00
38.05
A


ATOM
1114
N
ALA
A
153
28.098
42.094
−8.989
1.00
38.70
A


ATOM
1115
CA
ALA
A
153
29.106
43.145
−9.066
1.00
39.88
A


ATOM
1116
CB
ALA
A
153
29.783
43.117
−10.433
1.00
41.69
A


ATOM
1117
C
ALA
A
153
28.523
44.526
−8.805
1.00
40.13
A


ATOM
1118
O
ALA
A
153
29.120
45.342
−8.101
1.00
40.16
A


ATOM
1119
N
ARG
A
154
27.350
44.777
−9.374
1.00
40.05
A


ATOM
1120
CA
ARG
A
154
26.682
46.055
−9.217
1.00
40.63
A


ATOM
1121
CB
ARG
A
154
25.394
46.057
−10.044
1.00
42.51
A


ATOM
1122
CG
ARG
A
154
24.770
47.425
−10.221
1.00
47.49
A


ATOM
1123
CD
ARG
A
154
24.025
47.541
−11.555
1.00
48.14
A


ATOM
1124
NE
ARG
A
154
23.060
46.463
−11.764
1.00
48.78
A


ATOM
1125
CZ
ARG
A
154
23.298
45.368
−12.482
1.00
46.90
A


ATOM
1126
NH1
ARG
A
154
24.473
45.197
−13.069
1.00
48.01
A


ATOM
1127
NH2
ARG
A
154
22.359
44.443
−12.617
1.00
46.15
A


ATOM
1128
C
ARG
A
154
26.387
46.397
−7.750
1.00
40.64
A


ATOM
1129
O
ARG
A
154
26.297
47.574
−7.387
1.00
40.09
A


ATOM
1130
N
TYR
A
155
26.256
45.383
−6.897
1.00
40.35
A


ATOM
1131
CA
TYR
A
155
25.975
45.652
−5.493
1.00
38.19
A


ATOM
1132
CB
TYR
A
155
24.720
44.901
−5.052
1.00
38.67
A


ATOM
1133
CG
TYR
A
155
23.503
45.381
−5.790
1.00
38.24
A


ATOM
1134
CD1
TYR
A
155
23.171
44.855
−7.035
1.00
38.78
A


ATOM
1135
CE1
TYR
A
155
22.099
45.357
−7.766
1.00
39.06
A


ATOM
1136
CD2
TYR
A
155
22.725
46.424
−5.284
1.00
38.96
A


ATOM
1137
CE2
TYR
A
155
21.651
46.935
−6.006
1.00
40.27
A


ATOM
1138
CZ
TYR
A
155
21.346
46.398
−7.249
1.00
40.64
A


ATOM
1139
OH
TYR
A
155
20.301
46.910
−7.983
1.00
42.74
A


ATOM
1140
C
TYR
A
155
27.129
45.367
−4.545
1.00
38.20
A


ATOM
1141
O
TYR
A
155
26.957
45.356
−3.325
1.00
38.12
A


ATOM
1142
N
GLY
A
156
28.311
45.147
−5.105
1.00
37.37
A


ATOM
1143
CA
GLY
A
156
29.471
44.904
−4.270
1.00
37.64
A


ATOM
1144
C
GLY
A
156
29.602
43.510
−3.700
1.00
37.28
A


ATOM
1145
O
GLY
A
156
30.449
43.274
−2.835
1.00
37.51
A


ATOM
1146
N
LEU
A
157
28.765
42.586
−4.157
1.00
36.82
A


ATOM
1147
CA
LEU
A
157
28.846
41.210
−3.682
1.00
36.96
A


ATOM
1148
CB
LEU
A
157
27.531
40.480
−3.947
1.00
35.41
A


ATOM
1149
CG
LEU
A
157
26.314
41.101
−3.245
1.00
35.60
A


ATOM
1150
CD1
LEU
A
157
25.045
40.359
−3.646
1.00
33.80
A


ATOM
1151
CD2
LEU
A
157
26.508
41.048
−1.741
1.00
34.21
A


ATOM
1152
C
LEU
A
157
29.984
40.588
−4.479
1.00
37.62
A


ATOM
1153
O
LEU
A
157
29.814
40.233
−5.641
1.00
38.02
A


ATOM
1154
N
ASP
A
158
31.147
40.466
−3.847
1.00
37.12
A


ATOM
1155
CA
ASP
A
158
32.323
39.942
−4.526
1.00
37.41
A


ATOM
1156
CB
ASP
A
158
33.355
41.083
−4.669
1.00
38.84
A


ATOM
1157
CG
ASP
A
158
34.737
40.598
−5.106
1.00
39.06
A


ATOM
1158
OD1
ASP
A
158
34.823
39.688
−5.952
1.00
40.99
A


ATOM
1159
OD2
ASP
A
158
35.743
41.150
−4.610
1.00
42.10
A


ATOM
1160
C
ASP
A
158
32.948
38.713
−3.871
1.00
37.01
A


ATOM
1161
O
ASP
A
158
32.820
37.600
−4.386
1.00
37.39
A


ATOM
1162
N
LYS
A
159
33.608
38.902
−2.734
1.00
35.40
A


ATOM
1163
CA
LYS
A
159
34.258
37.790
−2.061
1.00
34.29
A


ATOM
1164
CB
LYS
A
159
35.313
38.316
−1.083
1.00
35.64
A


ATOM
1165
CG
LYS
A
159
36.464
39.047
−1.782
1.00
35.48
A


ATOM
1166
CD
LYS
A
159
37.021
38.202
−2.931
1.00
39.05
A


ATOM
1167
CE
LYS
A
159
38.115
38.924
−3.701
1.00
42.30
A


ATOM
1168
NZ
LYS
A
159
37.950
38.741
−5.171
1.00
44.17
A


ATOM
1169
C
LYS
A
159
33.312
36.824
−1.362
1.00
33.08
A


ATOM
1170
O
LYS
A
159
32.411
37.218
−0.615
1.00
32.50
A


ATOM
1171
N
VAL
A
160
33.555
35.546
−1.615
1.00
33.33
A


ATOM
1172
CA
VAL
A
160
32.764
34.453
−1.071
1.00
33.25
A


ATOM
1173
CB
VAL
A
160
32.317
33.542
−2.223
1.00
32.90
A


ATOM
1174
CG1
VAL
A
160
31.591
32.306
−1.698
1.00
33.69
A


ATOM
1175
CG2
VAL
A
160
31.429
34.344
−3.162
1.00
30.05
A


ATOM
1176
C
VAL
A
160
33.574
33.668
−0.045
1.00
33.60
A


ATOM
1177
O
VAL
A
160
34.608
33.086
−0.375
1.00
33.21
A


ATOM
1178
N
GLN
A
161
33.105
33.681
1.204
1.00
33.10
A


ATOM
1179
CA
GLN
A
161
33.760
32.981
2.308
1.00
34.17
A


ATOM
1180
CB
GLN
A
161
33.308
33.587
3.645
1.00
33.46
A


ATOM
1181
CG
GLN
A
161
33.870
32.894
4.882
1.00
38.50
A


ATOM
1182
CD
GLN
A
161
35.384
33.046
5.024
1.00
38.34
A


ATOM
1183
OE1
GLN
A
161
35.883
34.116
5.365
1.00
42.77
A


ATOM
1184
NE2
GLN
A
161
36.115
31.972
4.753
1.00
40.54
A


ATOM
1185
C
GLN
A
161
33.479
31.470
2.297
1.00
33.79
A


ATOM
1186
O
GLN
A
161
34.224
30.684
2.893
1.00
33.36
A


ATOM
1187
N
MET
A
162
32.401
31.065
1.629
1.00
31.86
A


ATOM
1188
CA
MET
A
162
32.056
29.649
1.536
1.00
30.83
A


ATOM
1189
CB
MET
A
162
31.812
29.048
2.925
1.00
34.42
A


ATOM
1190
CG
MET
A
162
30.424
29.337
3.461
1.00
36.50
A


ATOM
1191
SD
MET
A
162
30.433
30.247
5.003
1.00
43.69
A


ATOM
1192
CE
MET
A
162
29.357
29.180
5.978
1.00
40.85
A


ATOM
1193
C
MET
A
162
30.804
29.439
0.694
1.00
28.08
A


ATOM
1194
O
MET
A
162
30.068
30.373
0.416
1.00
29.84
A


ATOM
1195
N
THR
A
163
30.585
28.200
0.278
1.00
27.90
A


ATOM
1196
CA
THR
A
163
29.404
27.862
−0.501
1.00
28.23
A


ATOM
1197
CB
THR
A
163
29.753
27.471
−1.969
1.00
28.07
A


ATOM
1198
OG1
THR
A
163
30.534
26.268
−1.981
1.00
28.96
A


ATOM
1199
CG2
THR
A
163
30.515
28.579
−2.640
1.00
28.00
A


ATOM
1200
C
THR
A
163
28.781
26.657
0.160
1.00
28.16
A


ATOM
1201
O
THR
A
163
29.440
25.960
0.938
1.00
27.88
A


ATOM
1202
N
SER
A
164
27.500
26.426
−0.117
1.00
27.41
A


ATOM
1203
CA
SER
A
164
26.840
25.255
0.412
1.00
28.60
A


ATOM
1204
CB
SER
A
164
25.893
25.594
1.574
1.00
27.04
A


ATOM
1205
OG
SER
A
164
24.747
26.310
1.159
1.00
26.10
A


ATOM
1206
C
SER
A
164
26.078
24.620
−0.735
1.00
28.66
A


ATOM
1207
O
SER
A
164
25.693
25.301
−1.693
1.00
27.23
A


ATOM
1208
N
MET
A
165
25.909
23.306
−0.630
1.00
29.00
A


ATOM
1209
CA
MET
A
165
25.199
22.495
−1.606
1.00
31.67
A


ATOM
1210
CB
MET
A
165
26.157
21.497
−2.263
1.00
31.92
A


ATOM
1211
CG
MET
A
165
26.990
22.068
−3.396
1.00
35.54
A


ATOM
1212
SD
MET
A
165
28.324
23.135
−2.843
1.00
41.52
A


ATOM
1213
CE
MET
A
165
29.438
21.940
−2.325
1.00
31.41
A


ATOM
1214
C
MET
A
165
24.075
21.727
−0.920
1.00
29.72
A


ATOM
1215
O
MET
A
165
24.315
20.961
0.013
1.00
27.76
A


ATOM
1216
N
ASP
A
166
22.844
21.947
−1.375
1.00
30.14
A


ATOM
1217
CA
ASP
A
166
21.698
21.252
−0.814
1.00
29.25
A


ATOM
1218
CB
ASP
A
166
20.522
22.221
−0.637
1.00
31.09
A


ATOM
1219
CG
ASP
A
166
19.320
21.570
0.030
1.00
28.72
A


ATOM
1220
OD1
ASP
A
166
19.106
20.354
−0.163
1.00
28.28
A


ATOM
1221
OD2
ASP
A
166
18.580
22.284
0.733
1.00
30.52
A


ATOM
1222
C
ASP
A
166
21.359
20.167
−1.826
1.00
28.71
A


ATOM
1223
O
ASP
A
166
20.857
20.458
−2.918
1.00
27.49
A


ATOM
1224
N
TYR
A
167
21.650
18.923
−1.457
1.00
29.03
A


ATOM
1225
CA
TYR
A
167
21.433
17.766
−2.319
1.00
30.30
A


ATOM
1226
CB
TYR
A
167
22.224
16.577
−1.777
1.00
30.78
A


ATOM
1227
CG
TYR
A
167
23.710
16.837
−1.744
1.00
30.91
A


ATOM
1228
CD1
TYR
A
167
24.500
16.645
−2.889
1.00
29.35
A


ATOM
1229
CE1
TYR
A
167
25.854
16.960
−2.888
1.00
30.22
A


ATOM
1230
CD2
TYR
A
167
24.320
17.351
−0.597
1.00
28.48
A


ATOM
1231
CE2
TYR
A
167
25.676
17.672
−0.591
1.00
29.07
A


ATOM
1232
CZ
TYR
A
167
26.436
17.475
−1.735
1.00
28.34
A


ATOM
1233
OH
TYR
A
167
27.774
17.779
−1.732
1.00
28.84
A


ATOM
1234
C
TYR
A
167
19.974
17.371
−2.530
1.00
32.95
A


ATOM
1235
O
TYR
A
167
19.651
16.750
−3.531
1.00
31.97
A


ATOM
1236
N
LYS
A
168
19.096
17.730
−1.596
1.00
32.49
A


ATOM
1237
CA
LYS
A
168
17.678
17.408
−1.731
1.00
34.77
A


ATOM
1238
CB
LYS
A
168
17.017
17.311
−0.355
1.00
34.50
A


ATOM
1239
CG
LYS
A
168
17.505
16.127
0.448
1.00
35.07
A


ATOM
1240
CD
LYS
A
168
16.738
15.999
1.750
1.00
38.33
A


ATOM
1241
CE
LYS
A
168
17.052
14.688
2.438
1.00
39.81
A


ATOM
1242
NZ
LYS
A
168
16.297
14.561
3.714
1.00
41.97
A


ATOM
1243
C
LYS
A
168
16.937
18.427
−2.588
1.00
33.66
A


ATOM
1244
O
LYS
A
168
16.064
18.070
−3.368
1.00
35.97
A


ATOM
1245
N
LYS
A
169
17.285
19.696
−2.451
1.00
33.56
A


ATOM
1246
CA
LYS
A
169
16.623
20.720
−3.231
1.00
33.68
A


ATOM
1247
CB
LYS
A
169
16.364
21.946
−2.357
1.00
34.85
A


ATOM
1248
CG
LYS
A
169
15.481
21.625
−1.168
1.00
39.08
A


ATOM
1249
CD
LYS
A
169
15.060
22.866
−0.415
1.00
42.71
A


ATOM
1250
CE
LYS
A
169
14.149
22.503
0.749
1.00
44.29
A


ATOM
1251
NZ
LYS
A
169
13.666
23.718
1.473
1.00
47.87
A


ATOM
1252
C
LYS
A
169
17.424
21.113
−4.459
1.00
32.16
A


ATOM
1253
O
LYS
A
169
16.948
21.880
−5.291
1.00
32.91
A


ATOM
1254
N
ARG
A
170
18.635
20.573
−4.573
1.00
30.70
A


ATOM
1255
CA
ARG
A
170
19.519
20.889
−5.691
1.00
31.30
A


ATOM
1256
CB
ARG
A
170
18.959
20.322
−6.999
1.00
34.37
A


ATOM
1257
CG
ARG
A
170
18.911
18.794
−7.048
1.00
40.86
A


ATOM
1258
CD
ARG
A
170
18.230
18.309
−8.329
1.00
45.92
A


ATOM
1259
NE
ARG
A
170
18.162
16.848
−8.360
1.00
51.57
A


ATOM
1260
CZ
ARG
A
170
18.496
16.048
−7.350
1.00
53.83
A


ATOM
1261
NH1
ARG
A
170
18.930
16.558
−6.207
1.00
55.24
A


ATOM
1262
NH2
ARG
A
170
18.394
14.733
−7.489
1.00
55.41
A


ATOM
1263
C
ARG
A
170
19.684
22.403
−5.793
1.00
30.20
A


ATOM
1264
O
ARG
A
170
19.392
23.012
−6.827
1.00
27.37
A


ATOM
1265
N
GLN
A
171
20.157
23.006
−4.702
1.00
27.02
A


ATOM
1266
CA
GLN
A
171
20.361
24.446
−4.634
1.00
27.26
A


ATOM
1267
CB
GLN
A
171
19.271
25.112
−3.784
1.00
27.34
A


ATOM
1268
CG
GLN
A
171
17.861
24.995
−4.373
1.00
29.77
A


ATOM
1269
CD
GLN
A
171
16.795
25.663
−3.519
1.00
31.84
A


ATOM
1270
OE1
GLN
A
171
16.885
25.688
−2.293
1.00
30.82
A


ATOM
1271
NE2
GLN
A
171
15.762
26.192
−4.171
1.00
33.68
A


ATOM
1272
C
GLN
A
171
21.731
24.751
−4.044
1.00
26.95
A


ATOM
1273
O
GLN
A
171
22.286
23.940
−3.303
1.00
24.99
A


ATOM
1274
N
VAL
A
172
22.264
25.918
−4.397
1.00
26.41
A


ATOM
1275
CA
VAL
A
172
23.573
26.369
−3.936
1.00
29.58
A


ATOM
1276
CB
VAL
A
172
24.593
26.436
−5.114
1.00
27.24
A


ATOM
1277
CG1
VAL
A
172
25.931
26.995
−4.627
1.00
28.16
A


ATOM
1278
CG2
VAL
A
172
24.802
25.061
−5.703
1.00
27.97
A


ATOM
1279
C
VAL
A
172
23.476
27.761
−3.309
1.00
30.14
A


ATOM
1280
O
VAL
A
172
22.788
28.645
−3.836
1.00
32.05
A


ATOM
1281
N
ASN
A
173
24.153
27.941
−2.176
1.00
29.48
A


ATOM
1282
CA
ASN
A
173
24.182
29.228
−1.490
1.00
27.27
A


ATOM
1283
CB
ASN
A
173
23.887
29.074
0.014
1.00
29.16
A


ATOM
1284
CG
ASN
A
173
22.423
28.831
0.308
1.00
28.68
A


ATOM
1285
OD1
ASN
A
173
21.594
28.860
−0.595
1.00
27.97
A


ATOM
1286
ND2
ASN
A
173
22.094
28.599
1.591
1.00
26.44
A


ATOM
1287
C
ASN
A
173
25.588
29.809
−1.639
1.00
27.49
A


ATOM
1288
O
ASN
A
173
26.572
29.080
−1.523
1.00
27.57
A


ATOM
1289
N
LEU
A
174
25.672
31.109
−1.907
1.00
26.38
A


ATOM
1290
CA
LEU
A
174
26.961
31.793
−2.011
1.00
27.88
A


ATOM
1291
CB
LEU
A
174
27.047
32.605
−3.308
1.00
29.77
A


ATOM
1292
CG
LEU
A
174
26.831
31.854
−4.631
1.00
29.34
A


ATOM
1293
CD1
LEU
A
174
27.115
32.800
−5.784
1.00
30.72
A


ATOM
1294
CD2
LEU
A
174
27.750
30.647
−4.718
1.00
29.99
A


ATOM
1295
C
LEU
A
174
27.041
32.735
−0.810
1.00
27.33
A


ATOM
1296
O
LEU
A
174
26.324
33.737
−0.749
1.00
24.91
A


ATOM
1297
N
TYR
A
175
27.906
32.420
0.151
1.00
27.35
A


ATOM
1298
CA
TYR
A
175
28.023
33.266
1.330
1.00
28.53
A


ATOM
1299
CB
TYR
A
175
28.377
32.420
2.565
1.00
26.66
A


ATOM
1300
CG
TYR
A
175
27.343
31.347
2.890
1.00
28.27
A


ATOM
1301
CD1
TYR
A
175
27.339
30.115
2.229
1.00
28.80
A


ATOM
1302
CE1
TYR
A
175
26.352
29.150
2.496
1.00
29.04
A


ATOM
1303
CD2
TYR
A
175
26.340
31.588
3.826
1.00
29.47
A


ATOM
1304
CE2
TYR
A
175
25.355
30.642
4.094
1.00
29.90
A


ATOM
1305
CZ
TYR
A
175
25.360
29.432
3.433
1.00
30.82
A


ATOM
1306
OH
TYR
A
175
24.358
28.522
3.702
1.00
28.60
A


ATOM
1307
C
TYR
A
175
29.043
34.380
1.109
1.00
28.09
A


ATOM
1308
O
TYR
A
175
30.239
34.182
1.298
1.00
31.10
A


ATOM
1309
N
PHE
A
176
28.550
35.551
0.704
1.00
27.31
A


ATOM
1310
CA
PHE
A
176
29.393
36.714
0.444
1.00
28.26
A


ATOM
1311
CB
PHE
A
176
28.622
37.774
−0.341
1.00
27.28
A


ATOM
1312
CG
PHE
A
176
28.354
37.387
−1.772
1.00
27.93
A


ATOM
1313
CD1
PHE
A
176
27.081
36.992
−2.173
1.00
28.47
A


ATOM
1314
CD2
PHE
A
176
29.373
37.435
−2.719
1.00
28.88
A


ATOM
1315
CE1
PHE
A
176
26.822
36.650
−3.511
1.00
29.48
A


ATOM
1316
CE2
PHE
A
176
29.126
37.096
−4.062
1.00
28.83
A


ATOM
1317
CZ
PHE
A
176
27.848
36.704
−4.452
1.00
28.88
A


ATOM
1318
C
PHE
A
176
29.916
37.315
1.744
1.00
29.45
A


ATOM
1319
O
PHE
A
176
29.180
37.418
2.734
1.00
27.17
A


ATOM
1320
N
SER
A
177
31.184
37.726
1.722
1.00
30.00
A


ATOM
1321
CA
SER
A
177
31.841
38.281
2.910
1.00
31.18
A


ATOM
1322
CB
SER
A
177
32.876
37.270
3.419
1.00
30.63
A


ATOM
1323
OG
SER
A
177
33.179
37.472
4.786
1.00
31.80
A


ATOM
1324
C
SER
A
177
32.516
39.631
2.637
1.00
32.16
A


ATOM
1325
O
SER
A
177
32.466
40.149
1.513
1.00
31.15
A


ATOM
1326
N
GLU
A
178
33.161
40.188
3.663
1.00
32.64
A


ATOM
1327
CA
GLU
A
178
33.822
41.485
3.542
1.00
33.04
A


ATOM
1328
CB
GLU
A
178
35.134
41.352
2.743
1.00
35.78
A


ATOM
1329
CG
GLU
A
178
36.116
40.349
3.361
1.00
36.71
A


ATOM
1330
CD
GLU
A
178
37.519
40.402
2.759
1.00
37.67
A


ATOM
1331
OE1
GLU
A
178
37.654
40.565
1.521
1.00
36.25
A


ATOM
1332
OE2
GLU
A
178
38.490
40.259
3.535
1.00
37.28
A


ATOM
1333
C
GLU
A
178
32.866
42.458
2.856
1.00
33.20
A


ATOM
1334
O
GLU
A
178
33.208
43.106
1.860
1.00
35.37
A


ATOM
1335
N
LEU
A
179
31.653
42.548
3.395
1.00
32.06
A


ATOM
1336
CA
LEU
A
179
30.623
43.422
2.843
1.00
32.29
A


ATOM
1337
CB
LEU
A
179
29.261
43.035
3.417
1.00
31.59
A


ATOM
1338
CG
LEU
A
179
28.898
41.550
3.303
1.00
32.74
A


ATOM
1339
CD1
LEU
A
179
27.623
41.288
4.080
1.00
34.49
A


ATOM
1340
CD2
LEU
A
179
28.736
41.160
1.826
1.00
32.26
A


ATOM
1341
C
LEU
A
179
30.889
44.900
3.128
1.00
33.85
A


ATOM
1342
O
LEU
A
179
31.072
45.295
4.282
1.00
33.79
A


ATOM
1343
N
SER
A
180
30.905
45.717
2.078
1.00
34.71
A


ATOM
1344
CA
SER
A
180
31.132
47.149
2.240
1.00
36.87
A


ATOM
1345
CB
SER
A
180
31.390
47.821
0.890
1.00
38.51
A


ATOM
1346
OG
SER
A
180
30.177
48.062
0.205
1.00
41.84
A


ATOM
1347
C
SER
A
180
29.888
47.770
2.864
1.00
38.21
A


ATOM
1348
O
SER
A
180
28.784
47.236
2.723
1.00
35.22
A


ATOM
1349
N
ALA
A
181
30.073
48.898
3.544
1.00
37.72
A


ATOM
1350
CA
ALA
A
181
28.969
49.597
4.188
1.00
39.03
A


ATOM
1351
CB
ALA
A
181
29.487
50.855
4.886
1.00
37.87
A


ATOM
1352
C
ALA
A
181
27.901
49.968
3.166
1.00
39.40
A


ATOM
1353
O
ALA
A
181
26.701
49.909
3.449
1.00
39.97
A


ATOM
1354
N
GLN
A
182
28.342
50.337
1.971
1.00
38.70
A


ATOM
1355
CA
GLN
A
182
27.419
50.733
0.918
1.00
39.60
A


ATOM
1356
CB
GLN
A
182
28.192
51.136
−0.336
1.00
40.99
A


ATOM
1357
CG
GLN
A
182
27.303
51.450
−1.522
1.00
45.59
A


ATOM
1358
CD
GLN
A
182
27.915
52.474
−2.455
1.00
46.88
A


ATOM
1359
OE1
GLN
A
182
29.124
52.485
−2.683
1.00
51.04
A


ATOM
1360
NE2
GLN
A
182
27.075
53.336
−3.013
1.00
49.22
A


ATOM
1361
C
GLN
A
182
26.430
49.634
0.580
1.00
39.40
A


ATOM
1362
O
GLN
A
182
25.218
49.869
0.548
1.00
40.56
A


ATOM
1363
N
THR
A
183
26.951
48.435
0.330
1.00
38.08
A


ATOM
1364
CA
THR
A
183
26.125
47.284
−0.006
1.00
36.74
A


ATOM
1365
CB
THR
A
183
26.980
45.990
−0.097
1.00
36.40
A


ATOM
1366
OG1
THR
A
183
27.950
46.122
−1.146
1.00
36.47
A


ATOM
1367
CG2
THR
A
183
26.098
44.789
−0.378
1.00
34.06
A


ATOM
1368
C
THR
A
183
25.003
47.045
1.005
1.00
36.17
A


ATOM
1369
O
THR
A
183
23.900
46.661
0.627
1.00
35.97
A


ATOM
1370
N
LEU
A
184
25.282
47.278
2.284
1.00
35.23
A


ATOM
1371
CA
LEU
A
184
24.287
47.051
3.327
1.00
35.08
A


ATOM
1372
CB
LEU
A
184
24.969
46.532
4.598
1.00
34.37
A


ATOM
1373
CG
LEU
A
184
25.642
45.161
4.475
1.00
32.38
A


ATOM
1374
CD1
LEU
A
184
26.367
44.840
5.767
1.00
34.20
A


ATOM
1375
CD2
LEU
A
184
24.609
44.089
4.153
1.00
31.86
A


ATOM
1376
C
LEU
A
184
23.397
48.241
3.681
1.00
37.71
A


ATOM
1377
O
LEU
A
184
22.552
48.128
4.574
1.00
36.83
A


ATOM
1378
N
GLU
A
185
23.577
49.375
3.007
1.00
37.02
A


ATOM
1379
CA
GLU
A
185
22.726
50.525
3.296
1.00
39.60
A


ATOM
1380
CB
GLU
A
185
23.325
51.831
2.771
1.00
42.09
A


ATOM
1381
CG
GLU
A
185
24.105
52.608
3.819
1.00
47.09
A


ATOM
1382
CD
GLU
A
185
23.375
52.699
5.158
1.00
49.21
A


ATOM
1383
OE1
GLU
A
185
22.140
52.931
5.168
1.00
50.57
A


ATOM
1384
OE2
GLU
A
185
24.040
52.546
6.206
1.00
48.86
A


ATOM
1385
C
GLU
A
185
21.369
50.296
2.660
1.00
37.48
A


ATOM
1386
O
GLU
A
185
21.261
49.592
1.656
1.00
35.55
A


ATOM
1387
N
ALA
A
186
20.342
50.898
3.257
1.00
37.32
A


ATOM
1388
CA
ALA
A
186
18.965
50.754
2.795
1.00
35.80
A


ATOM
1389
CB
ALA
A
186
18.073
51.738
3.535
1.00
35.92
A


ATOM
1390
C
ALA
A
186
18.777
50.915
1.290
1.00
36.43
A


ATOM
1391
O
ALA
A
186
18.143
50.080
0.645
1.00
34.72
A


ATOM
1392
N
GLU
A
187
19.316
51.995
0.734
1.00
37.69
A


ATOM
1393
CA
GLU
A
187
19.183
52.254
−0.690
1.00
38.57
A


ATOM
1394
CB
GLU
A
187
19.920
53.549
−1.042
1.00
42.29
A


ATOM
1395
CG
GLU
A
187
20.121
53.794
−2.529
1.00
46.35
A


ATOM
1396
CD
GLU
A
187
20.874
55.089
−2.798
1.00
48.77
A


ATOM
1397
OE1
GLU
A
187
20.265
56.173
−2.642
1.00
51.07
A


ATOM
1398
OE2
GLU
A
187
22.075
55.019
−3.149
1.00
49.51
A


ATOM
1399
C
GLU
A
187
19.697
51.092
−1.536
1.00
37.10
A


ATOM
1400
O
GLU
A
187
19.036
50.672
−2.482
1.00
36.07
A


ATOM
1401
N
SER
A
188
20.874
50.580
−1.195
1.00
36.10
A


ATOM
1402
CA
SER
A
188
21.476
49.461
−1.914
1.00
34.79
A


ATOM
1403
CB
SER
A
188
22.916
49.240
−1.439
1.00
35.13
A


ATOM
1404
OG
SER
A
188
23.412
47.986
−1.888
1.00
42.19
A


ATOM
1405
C
SER
A
188
20.685
48.161
−1.749
1.00
33.67
A


ATOM
1406
O
SER
A
188
20.430
47.450
−2.725
1.00
32.43
A


ATOM
1407
N
VAL
A
189
20.308
47.846
−0.512
1.00
32.53
A


ATOM
1408
CA
VAL
A
189
19.548
46.633
−0.241
1.00
33.25
A


ATOM
1409
CB
VAL
A
189
19.229
46.498
1.269
1.00
34.36
A


ATOM
1410
CG1
VAL
A
189
18.267
45.345
1.500
1.00
32.73
A


ATOM
1411
CG2
VAL
A
189
20.512
46.259
2.052
1.00
35.61
A


ATOM
1412
C
VAL
A
189
18.240
46.610
−1.032
1.00
32.13
A


ATOM
1413
O
VAL
A
189
17.916
45.611
−1.660
1.00
31.84
A


ATOM
1414
N
LEU
A
190
17.495
47.714
−1.005
1.00
32.08
A


ATOM
1415
CA
LEU
A
190
16.220
47.798
−1.718
1.00
32.32
A


ATOM
1416
CB
LEU
A
190
15.539
49.139
−1.426
1.00
33.11
A


ATOM
1417
CG
LEU
A
190
14.876
49.287
−0.052
1.00
32.87
A


ATOM
1418
CD1
LEU
A
190
14.378
50.724
0.120
1.00
33.09
A


ATOM
1419
CD2
LEU
A
190
13.713
48.304
0.071
1.00
31.45
A


ATOM
1420
C
LEU
A
190
16.362
47.618
−3.229
1.00
33.42
A


ATOM
1421
O
LEU
A
190
15.555
46.936
−3.855
1.00
33.09
A


ATOM
1422
N
ALA
A
191
17.379
48.238
−3.816
1.00
33.70
A


ATOM
1423
CA
ALA
A
191
17.600
48.113
−5.251
1.00
34.39
A


ATOM
1424
CB
ALA
A
191
18.766
48.987
−5.681
1.00
36.16
A


ATOM
1425
C
ALA
A
191
17.886
46.656
−5.576
1.00
35.14
A


ATOM
1426
O
ALA
A
191
17.399
46.122
−6.572
1.00
36.27
A


ATOM
1427
N
LEU
A
192
18.674
46.012
−4.718
1.00
36.49
A


ATOM
1428
CA
LEU
A
192
19.023
44.612
−4.902
1.00
34.74
A


ATOM
1429
CB
LEU
A
192
20.024
44.165
−3.830
1.00
34.60
A


ATOM
1430
CG
LEU
A
192
20.375
42.671
−3.754
1.00
34.43
A


ATOM
1431
CD1
LEU
A
192
20.924
42.176
−5.084
1.00
33.47
A


ATOM
1432
CD2
LEU
A
192
21.394
42.451
−2.645
1.00
34.05
A


ATOM
1433
C
LEU
A
192
17.782
43.727
−4.858
1.00
35.29
A


ATOM
1434
O
LEU
A
192
17.570
42.927
−5.767
1.00
33.59
A


ATOM
1435
N
VAL
A
193
16.962
43.865
−3.816
1.00
35.74
A


ATOM
1436
CA
VAL
A
193
15.763
43.039
−3.717
1.00
38.00
A


ATOM
1437
CB
VAL
A
193
14.985
43.274
−2.395
1.00
37.63
A


ATOM
1438
CG1
VAL
A
193
15.920
43.105
−1.201
1.00
39.87
A


ATOM
1439
CG2
VAL
A
193
14.351
44.634
−2.389
1.00
37.77
A


ATOM
1440
C
VAL
A
193
14.837
43.327
−4.887
1.00
38.50
A


ATOM
1441
O
VAL
A
193
14.280
42.413
−5.483
1.00
38.16
A


ATOM
1442
N
ARG
A
194
14.677
44.604
−5.207
1.00
40.37
A


ATOM
1443
CA
ARG
A
194
13.830
45.019
−6.314
1.00
43.28
A


ATOM
1444
CB
ARG
A
194
13.899
46.549
−6.461
1.00
45.33
A


ATOM
1445
CG
ARG
A
194
12.637
47.237
−6.974
1.00
50.72
A


ATOM
1446
CD
ARG
A
194
12.881
48.130
−8.183
1.00
54.41
A


ATOM
1447
NE
ARG
A
194
11.661
48.393
−8.942
1.00
57.37
A


ATOM
1448
CZ
ARG
A
194
10.458
48.505
−8.389
1.00
59.05
A


ATOM
1449
NH1
ARG
A
194
10.317
48.368
−7.079
1.00
61.10
A


ATOM
1450
NH2
ARG
A
194
9.398
48.773
−9.140
1.00
61.23
A


ATOM
1451
C
ARG
A
194
14.283
44.305
−7.588
1.00
41.65
A


ATOM
1452
O
ARG
A
194
13.481
43.683
−8.280
1.00
41.71
A


ATOM
1453
N
GLU
A
195
15.576
44.360
−7.877
1.00
41.34
A


ATOM
1454
CA
GLU
A
195
16.094
43.713
−9.070
1.00
41.44
A


ATOM
1455
CB
GLU
A
195
17.565
44.071
−9.264
1.00
43.69
A


ATOM
1456
CG
GLU
A
195
17.882
44.479
−10.693
1.00
46.31
A


ATOM
1457
CD
GLU
A
195
19.346
44.778
−10.911
1.00
45.36
A


ATOM
1458
OE1
GLU
A
195
19.879
45.680
−10.233
1.00
45.90
A


ATOM
1459
OE2
GLU
A
195
19.960
44.107
−11.765
1.00
46.65
A


ATOM
1460
C
GLU
A
195
15.925
42.189
−9.052
1.00
41.97
A


ATOM
1461
O
GLU
A
195
15.700
41.577
−10.097
1.00
41.48
A


ATOM
1462
N
LEU
A
196
16.032
41.576
−7.874
1.00
40.20
A


ATOM
1463
CA
LEU
A
196
15.878
40.126
−7.762
1.00
38.53
A


ATOM
1464
CB
LEU
A
196
16.686
39.590
−6.570
1.00
36.27
A


ATOM
1465
CG
LEU
A
196
18.212
39.731
−6.649
1.00
37.41
A


ATOM
1466
CD1
LEU
A
196
18.852
39.207
−5.371
1.00
36.85
A


ATOM
1467
CD2
LEU
A
196
18.740
38.969
−7.855
1.00
37.56
A


ATOM
1468
C
LEU
A
196
14.399
39.753
−7.608
1.00
38.13
A


ATOM
1469
O
LEU
A
196
14.047
38.576
−7.509
1.00
38.58
A


ATOM
1470
N
GLY
A
197
13.541
40.768
−7.590
1.00
38.66
A


ATOM
1471
CA
GLY
A
197
12.112
40.537
−7.458
1.00
38.24
A


ATOM
1472
C
GLY
A
197
11.712
39.891
−6.149
1.00
38.25
A


ATOM
1473
O
GLY
A
197
10.833
39.028
−6.121
1.00
38.47
A


ATOM
1474
N
LEU
A
198
12.334
40.318
−5.055
1.00
37.09
A


ATOM
1475
CA
LEU
A
198
12.031
39.733
−3.756
1.00
36.88
A


ATOM
1476
CB
LEU
A
198
13.340
39.324
−3.065
1.00
35.42
A


ATOM
1477
CG
LEU
A
198
14.267
38.410
−3.874
1.00
36.31
A


ATOM
1478
CD1
LEU
A
198
15.612
38.284
−3.170
1.00
37.54
A


ATOM
1479
CD2
LEU
A
198
13.631
37.037
−4.050
1.00
36.41
A


ATOM
1480
C
LEU
A
198
11.211
40.629
−2.827
1.00
36.62
A


ATOM
1481
O
LEU
A
198
11.042
41.829
−3.064
1.00
33.54
A


ATOM
1482
N
HIS
A
199
10.707
40.025
−1.759
1.00
36.58
A


ATOM
1483
CA
HIS
A
199
9.918
40.738
−0.763
1.00
37.83
A


ATOM
1484
CB
HIS
A
199
9.667
39.833
0.440
1.00
39.68
A


ATOM
1485
CG
HIS
A
199
8.795
40.447
1.487
1.00
39.54
A


ATOM
1486
CD2
HIS
A
199
9.094
40.986
2.692
1.00
41.01
A


ATOM
1487
ND1
HIS
A
199
7.429
40.549
1.348
1.00
42.14
A


ATOM
1488
CE1
HIS
A
199
6.921
41.121
2.426
1.00
42.34
A


ATOM
1489
NE2
HIS
A
199
7.911
41.396
3.257
1.00
43.77
A


ATOM
1490
C
HIS
A
199
10.650
41.992
−0.296
1.00
38.39
A


ATOM
1491
O
HIS
A
199
11.845
41.950
0.012
1.00
38.41
A


ATOM
1492
N
VAL
A
200
9.928
43.107
−0.246
1.00
37.60
A


ATOM
1493
CA
VAL
A
200
10.506
44.367
0.188
1.00
37.98
A


ATOM
1494
CB
VAL
A
200
9.689
45.569
−0.348
1.00
38.84
A


ATOM
1495
CG1
VAL
A
200
10.254
46.869
0.201
1.00
39.30
A


ATOM
1496
CG2
VAL
A
200
9.709
45.578
−1.878
1.00
39.44
A


ATOM
1497
C
VAL
A
200
10.537
44.447
1.714
1.00
38.23
A


ATOM
1498
O
VAL
A
200
9.506
44.353
2.369
1.00
37.38
A


ATOM
1499
N
PRO
A
201
11.734
44.604
2.302
1.00
38.40
A


ATOM
1500
CD
PRO
A
201
13.070
44.490
1.687
1.00
36.88
A


ATOM
1501
CA
PRO
A
201
11.835
44.694
3.761
1.00
38.03
A


ATOM
1502
CB
PRO
A
201
13.306
44.377
4.012
1.00
39.05
A


ATOM
1503
CG
PRO
A
201
13.971
44.949
2.792
1.00
37.38
A


ATOM
1504
C
PRO
A
201
11.436
46.091
4.252
1.00
39.00
A


ATOM
1505
O
PRO
A
201
11.552
47.068
3.516
1.00
38.29
A


ATOM
1506
N
ASN
A
202
10.947
46.191
5.483
1.00
38.73
A


ATOM
1507
CA
ASN
A
202
10.564
47.492
6.007
1.00
38.53
A


ATOM
1508
CB
ASN
A
202
9.273
47.393
6.827
1.00
37.55
A


ATOM
1509
CG
ASN
A
202
9.323
46.302
7.861
1.00
36.97
A


ATOM
1510
OD1
ASN
A
202
10.368
46.051
8.463
1.00
34.21
A


ATOM
1511
ND2
ASN
A
202
8.183
45.650
8.090
1.00
35.55
A


ATOM
1512
C
ASN
A
202
11.687
48.072
6.857
1.00
39.15
A


ATOM
1513
O
ASN
A
202
12.798
47.539
6.877
1.00
38.30
A


ATOM
1514
N
GLU
A
203
11.392
49.161
7.561
1.00
38.76
A


ATOM
1515
CA
GLU
A
203
12.386
49.835
8.391
1.00
41.10
A


ATOM
1516
CB
GLU
A
203
11.727
50.944
9.212
1.00
44.04
A


ATOM
1517
CG
GLU
A
203
12.689
52.040
9.638
1.00
49.57
A


ATOM
1518
CD
GLU
A
203
12.048
53.040
10.584
1.00
52.76
A


ATOM
1519
OE1
GLU
A
203
10.873
53.402
10.362
1.00
54.28
A


ATOM
1520
OE2
GLU
A
203
12.722
53.470
11.543
1.00
54.88
A


ATOM
1521
C
GLU
A
203
13.128
48.889
9.328
1.00
39.75
A


ATOM
1522
O
GLU
A
203
14.353
48.941
9.434
1.00
38.21
A


ATOM
1523
N
LEU
A
204
12.383
48.028
10.006
1.00
38.08
A


ATOM
1524
CA
LEU
A
204
12.984
47.081
10.932
1.00
37.43
A


ATOM
1525
CB
LEU
A
204
11.884
46.297
11.649
1.00
39.35
A


ATOM
1526
CG
LEU
A
204
12.342
45.230
12.644
1.00
41.12
A


ATOM
1527
CD1
LEU
A
204
13.153
45.882
13.753
1.00
41.94
A


ATOM
1528
CD2
LEU
A
204
11.131
44.512
13.218
1.00
43.55
A


ATOM
1529
C
LEU
A
204
13.922
46.121
10.195
1.00
37.20
A


ATOM
1530
O
LEU
A
204
15.060
45.897
10.624
1.00
36.41
A


ATOM
1531
N
GLY
A
205
13.438
45.561
9.086
1.00
34.65
A


ATOM
1532
CA
GLY
A
205
14.240
44.634
8.303
1.00
35.07
A


ATOM
1533
C
GLY
A
205
15.512
45.257
7.756
1.00
34.00
A


ATOM
1534
O
GLY
A
205
16.577
44.628
7.753
1.00
33.65
A


ATOM
1535
N
LEU
A
206
15.408
46.500
7.297
1.00
34.66
A


ATOM
1536
CA
LEU
A
206
16.559
47.201
6.734
1.00
35.09
A


ATOM
1537
CB
LEU
A
206
16.095
48.464
6.012
1.00
36.03
A


ATOM
1538
CG
LEU
A
206
15.387
48.179
4.679
1.00
33.49
A


ATOM
1539
CD1
LEU
A
206
14.670
49.426
4.182
1.00
33.05
A


ATOM
1540
CD2
LEU
A
206
16.404
47.701
3.662
1.00
32.40
A


ATOM
1541
C
LEU
A
206
17.616
47.532
7.782
1.00
36.87
A


ATOM
1542
O
LEU
A
206
18.821
47.468
7.505
1.00
36.00
A


ATOM
1543
N
LYS
A
207
17.168
47.878
8.987
1.00
36.83
A


ATOM
1544
CA
LYS
A
207
18.091
48.177
10.074
1.00
37.90
A


ATOM
1545
CB
LYS
A
207
17.320
48.600
11.326
1.00
39.99
A


ATOM
1546
CG
LYS
A
207
18.216
49.007
12.495
1.00
45.04
A


ATOM
1547
CD
LYS
A
207
17.400
49.347
13.740
1.00
50.01
A


ATOM
1548
CE
LYS
A
207
18.268
49.988
14.828
1.00
52.95
A


ATOM
1549
NZ
LYS
A
207
19.403
49.110
15.254
1.00
55.35
A


ATOM
1550
C
LYS
A
207
18.891
46.903
10.362
1.00
37.50
A


ATOM
1551
O
LYS
A
207
20.095
46.951
10.611
1.00
38.54
A


ATOM
1552
N
PHE
A
208
18.208
45.762
10.312
1.00
35.87
A


ATOM
1553
CA
PHE
A
208
18.831
44.468
10.552
1.00
35.98
A


ATOM
1554
CB
PHE
A
208
17.746
43.376
10.613
1.00
34.44
A


ATOM
1555
CG
PHE
A
208
18.288
41.966
10.743
1.00
36.93
A


ATOM
1556
CD1
PHE
A
208
18.732
41.264
9.621
1.00
35.75
A


ATOM
1557
CD2
PHE
A
208
18.328
41.332
11.987
1.00
36.14
A


ATOM
1558
CE1
PHE
A
208
19.201
39.955
9.734
1.00
36.55
A


ATOM
1559
CE2
PHE
A
208
18.798
40.020
12.110
1.00
37.86
A


ATOM
1560
CZ
PHE
A
208
19.234
39.330
10.977
1.00
37.36
A


ATOM
1561
C
PHE
A
208
19.866
44.149
9.464
1.00
35.26
A


ATOM
1562
O
PHE
A
208
20.976
43.700
9.758
1.00
34.23
A


ATOM
1563
N
CYS
A
209
19.508
44.389
8.208
1.00
35.13
A


ATOM
1564
CA
CYS
A
209
20.424
44.115
7.106
1.00
35.37
A


ATOM
1565
CB
CYS
A
209
19.756
44.438
5.762
1.00
33.36
A


ATOM
1566
SG
CYS
A
209
18.357
43.364
5.322
1.00
33.51
A


ATOM
1567
C
CYS
A
209
21.720
44.920
7.244
1.00
35.58
A


ATOM
1568
O
CYS
A
209
22.805
44.416
6.961
1.00
36.94
A


ATOM
1569
N
LYS
A
210
21.592
46.172
7.669
1.00
35.97
A


ATOM
1570
CA
LYS
A
210
22.736
47.067
7.844
1.00
36.87
A


ATOM
1571
CB
LYS
A
210
22.239
48.429
8.341
1.00
37.10
A


ATOM
1572
CG
LYS
A
210
23.296
49.330
8.973
1.00
42.31
A


ATOM
1573
CD
LYS
A
210
22.677
50.663
9.409
1.00
44.14
A


ATOM
1574
CE
LYS
A
210
23.705
51.801
9.307
1.00
47.02
A


ATOM
1575
NZ
LYS
A
210
23.114
53.178
9.361
1.00
48.89
A


ATOM
1576
C
LYS
A
210
23.761
46.497
8.824
1.00
36.21
A


ATOM
1577
O
LYS
A
210
24.949
46.793
8.739
1.00
37.65
A


ATOM
1578
N
ARG
A
211
23.285
45.669
9.743
1.00
35.86
A


ATOM
1579
CA
ARG
A
211
24.123
45.053
10.765
1.00
36.74
A


ATOM
1580
CB
ARG
A
211
23.273
44.863
12.023
1.00
37.38
A


ATOM
1581
CG
ARG
A
211
23.965
44.280
13.223
1.00
40.40
A


ATOM
1582
CD
ARG
A
211
23.039
44.393
14.423
1.00
43.85
A


ATOM
1583
NE
ARG
A
211
23.632
43.875
15.651
1.00
46.63
A


ATOM
1584
CZ
ARG
A
211
23.104
44.050
16.857
1.00
48.65
A


ATOM
1585
NH1
ARG
A
211
21.973
44.734
16.992
1.00
49.90
A


ATOM
1586
NH2
ARG
A
211
23.697
43.535
17.929
1.00
48.82
A


ATOM
1587
C
ARG
A
211
24.735
43.715
10.325
1.00
35.55
A


ATOM
1588
O
ARG
A
211
25.559
43.141
11.038
1.00
35.83
A


ATOM
1589
N
SER
A
212
24.345
43.238
9.144
1.00
33.22
A


ATOM
1590
CA
SER
A
212
24.818
41.954
8.607
1.00
33.87
A


ATOM
1591
CB
SER
A
212
24.125
41.652
7.269
1.00
31.55
A


ATOM
1592
OG
SER
A
212
22.738
41.403
7.448
1.00
33.21
A


ATOM
1593
C
SER
A
212
26.319
41.789
8.401
1.00
32.15
A


ATOM
1594
O
SER
A
212
27.009
42.714
8.009
1.00
35.39
A


ATOM
1595
N
PHE
A
213
26.818
40.585
8.657
1.00
33.61
A


ATOM
1596
CA
PHE
A
213
28.236
40.295
8.447
1.00
34.25
A


ATOM
1597
CB
PHE
A
213
28.862
39.641
9.695
1.00
33.84
A


ATOM
1598
CG
PHE
A
213
28.500
38.192
9.887
1.00
34.55
A


ATOM
1599
CD1
PHE
A
213
29.384
37.186
9.509
1.00
34.18
A


ATOM
1600
CD2
PHE
A
213
27.291
37.834
10.471
1.00
34.89
A


ATOM
1601
CE1
PHE
A
213
29.071
35.846
9.717
1.00
37.02
A


ATOM
1602
CE2
PHE
A
213
26.965
36.501
10.683
1.00
36.59
A


ATOM
1603
CZ
PHE
A
213
27.858
35.500
10.306
1.00
35.85
A


ATOM
1604
C
PHE
A
213
28.349
39.363
7.242
1.00
33.28
A


ATOM
1605
O
PHE
A
213
29.422
39.178
6.685
1.00
32.96
A


ATOM
1606
N
SER
A
214
27.222
38.786
6.838
1.00
34.50
A


ATOM
1607
CA
SER
A
214
27.192
37.874
5.698
1.00
31.86
A


ATOM
1608
CB
SER
A
214
27.375
36.434
6.170
1.00
33.79
A


ATOM
1609
OG
SER
A
214
27.224
35.521
5.093
1.00
37.93
A


ATOM
1610
C
SER
A
214
25.860
37.988
4.974
1.00
31.77
A


ATOM
1611
O
SER
A
214
24.833
38.202
5.610
1.00
28.63
A


ATOM
1612
N
VAL
A
215
25.892
37.856
3.648
1.00
30.90
A


ATOM
1613
CA
VAL
A
215
24.685
37.908
2.818
1.00
29.86
A


ATOM
1614
CB
VAL
A
215
24.618
39.187
1.947
1.00
30.29
A


ATOM
1615
CG1
VAL
A
215
23.443
39.103
0.993
1.00
32.52
A


ATOM
1616
CG2
VAL
A
215
24.460
40.412
2.817
1.00
33.53
A


ATOM
1617
C
VAL
A
215
24.759
36.725
1.866
1.00
28.93
A


ATOM
1618
O
VAL
A
215
25.802
36.513
1.238
1.00
28.44
A


ATOM
1619
N
TYR
A
216
23.679
35.952
1.749
1.00
27.25
A


ATOM
1620
CA
TYR
A
216
23.703
34.817
0.824
1.00
27.51
A


ATOM
1621
CB
TYR
A
216
24.119
33.499
1.522
1.00
26.91
A


ATOM
1622
CG
TYR
A
216
23.361
33.072
2.774
1.00
27.45
A


ATOM
1623
CD1
TYR
A
216
22.508
31.959
2.769
1.00
26.55
A


ATOM
1624
CE1
TYR
A
216
21.913
31.487
3.967
1.00
29.46
A


ATOM
1625
CD2
TYR
A
216
23.593
33.710
3.992
1.00
30.60
A


ATOM
1626
CE2
TYR
A
216
23.016
33.258
5.178
1.00
29.33
A


ATOM
1627
CZ
TYR
A
216
22.186
32.156
5.165
1.00
29.32
A


ATOM
1628
OH
TYR
A
216
21.654
31.734
6.360
1.00
31.96
A


ATOM
1629
C
TYR
A
216
22.477
34.534
−0.019
1.00
27.43
A


ATOM
1630
O
TYR
A
216
21.383
34.337
0.492
1.00
25.60
A


ATOM
1631
N
PRO
A
217
22.658
34.519
−1.349
1.00
28.06
A


ATOM
1632
CD
PRO
A
217
23.839
35.061
−2.054
1.00
28.05
A


ATOM
1633
CA
PRO
A
217
21.576
34.241
−2.293
1.00
27.27
A


ATOM
1634
CB
PRO
A
217
22.008
35.008
−3.534
1.00
28.73
A


ATOM
1635
CG
PRO
A
217
23.512
34.790
−3.521
1.00
27.12
A


ATOM
1636
C
PRO
A
217
21.558
32.736
−2.550
1.00
27.30
A


ATOM
1637
O
PRO
A
217
22.596
32.066
−2.432
1.00
26.63
A


ATOM
1638
N
THR
A
218
20.393
32.204
−2.890
1.00
25.62
A


ATOM
1639
CA
THR
A
218
20.258
30.784
−3.204
1.00
26.97
A


ATOM
1640
CB
THR
A
218
19.073
30.132
−2.452
1.00
27.35
A


ATOM
1641
OG1
THR
A
218
19.274
30.242
−1.035
1.00
28.53
A


ATOM
1642
CG2
THR
A
218
18.962
28.645
−2.818
1.00
26.30
A


ATOM
1643
C
THR
A
218
19.991
30.674
−4.707
1.00
28.41
A


ATOM
1644
O
THR
A
218
19.115
31.364
−5.231
1.00
29.31
A


ATOM
1645
N
LEU
A
219
20.755
29.823
−5.388
1.00
29.45
A


ATOM
1646
CA
LEU
A
219
20.608
29.619
−6.826
1.00
31.35
A


ATOM
1647
CB
LEU
A
219
21.884
30.066
−7.562
1.00
32.22
A


ATOM
1648
CG
LEU
A
219
22.337
31.509
−7.312
1.00
32.16
A


ATOM
1649
CD1
LEU
A
219
23.346
31.516
−6.160
1.00
31.96
A


ATOM
1650
CD2
LEU
A
219
22.961
32.105
−8.584
1.00
29.71
A


ATOM
1651
C
LEU
A
219
20.315
28.149
−7.148
1.00
32.09
A


ATOM
1652
O
LEU
A
219
20.540
27.255
−6.329
1.00
31.23
A


ATOM
1653
N
ASN
A
220
19.812
27.898
−8.349
1.00
33.47
A


ATOM
1654
CA
ASN
A
220
19.498
26.537
−8.767
1.00
34.71
A


ATOM
1655
CB
ASN
A
220
17.990
26.289
−8.648
1.00
36.03
A


ATOM
1656
CG
ASN
A
220
17.186
27.115
−9.629
1.00
39.61
A


ATOM
1657
OD1
ASN
A
220
17.141
26.809
−10.819
1.00
40.74
A


ATOM
1658
ND2
ASN
A
220
16.558
28.177
−9.138
1.00
38.72
A


ATOM
1659
C
ASN
A
220
19.965
26.294
−10.205
1.00
35.49
A


ATOM
1660
O
ASN
A
220
20.354
27.228
−10.906
1.00
33.83
A


ATOM
1661
N
TRP
A
221
19.927
25.033
−10.623
1.00
36.36
A


ATOM
1662
CA
TRP
A
221
20.354
24.615
−11.956
1.00
38.77
A


ATOM
1663
CB
TRP
A
221
20.845
23.162
−11.906
1.00
37.13
A


ATOM
1664
CG
TRP
A
221
22.255
22.982
−11.415
1.00
35.49
A


ATOM
1665
CD2
TRP
A
221
22.679
22.785
−10.056
1.00
34.23
A


ATOM
1666
CE2
TRP
A
221
24.084
22.613
−10.078
1.00
34.61
A


ATOM
1667
CE3
TRP
A
221
22.009
22.733
−8.826
1.00
33.47
A


ATOM
1668
CD1
TRP
A
221
23.388
22.932
−12.180
1.00
36.61
A


ATOM
1669
NE1
TRP
A
221
24.490
22.707
−11.384
1.00
36.62
A


ATOM
1670
CZ2
TRP
A
221
24.832
22.391
−8.916
1.00
31.63
A


ATOM
1671
CZ3
TRP
A
221
22.752
22.511
−7.665
1.00
31.82
A


ATOM
1672
CH2
TRP
A
221
24.153
22.343
−7.722
1.00
33.18
A


ATOM
1673
C
TRP
A
221
19.261
24.736
−13.020
1.00
41.28
A


ATOM
1674
O
TRP
A
221
19.546
24.642
−14.214
1.00
41.24
A


ATOM
1675
N
GLU
A
222
18.017
24.943
−12.594
1.00
42.94
A


ATOM
1676
CA
GLU
A
222
16.905
25.058
−13.534
1.00
46.16
A


ATOM
1677
CB
GLU
A
222
15.587
24.689
−12.848
1.00
46.48
A


ATOM
1678
CG
GLU
A
222
15.530
23.273
−12.303
1.00
49.88
A


ATOM
1679
CD
GLU
A
222
15.994
22.238
−13.307
1.00
52.45
A


ATOM
1680
OE1
GLU
A
222
15.705
22.404
−14.512
1.00
55.71
A


ATOM
1681
OE2
GLU
A
222
16.640
21.251
−12.894
1.00
53.80
A


ATOM
1682
C
GLU
A
222
16.748
26.425
−14.203
1.00
46.68
A


ATOM
1683
O
GLU
A
222
16.463
26.501
−15.398
1.00
47.48
A


ATOM
1684
N
THR
A
223
16.920
27.500
−13.439
1.00
46.30
A


ATOM
1685
CA
THR
A
223
16.777
28.848
−13.985
1.00
46.13
A


ATOM
1686
CB
THR
A
223
15.407
29.462
−13.613
1.00
47.72
A


ATOM
1687
OG1
THR
A
223
15.332
29.646
−12.194
1.00
49.52
A


ATOM
1688
CG2
THR
A
223
14.270
28.546
−14.051
1.00
49.10
A


ATOM
1689
C
THR
A
223
17.867
29.791
−13.477
1.00
46.02
A


ATOM
1690
O
THR
A
223
18.643
29.437
−12.591
1.00
44.84
A


ATOM
1691
N
GLY
A
224
17.915
30.994
−14.044
1.00
44.46
A


ATOM
1692
CA
GLY
A
224
18.906
31.971
−13.626
1.00
43.49
A


ATOM
1693
C
GLY
A
224
18.341
32.872
−12.545
1.00
42.65
A


ATOM
1694
O
GLY
A
224
19.014
33.777
−12.057
1.00
43.27
A


ATOM
1695
N
LYS
A
225
17.093
32.618
−12.167
1.00
41.43
A


ATOM
1696
CA
LYS
A
225
16.426
33.415
−11.150
1.00
41.27
A


ATOM
1697
CB
LYS
A
225
14.908
33.310
−11.320
1.00
43.63
A


ATOM
1698
CG
LYS
A
225
14.111
34.119
−10.308
1.00
47.97
A


ATOM
1699
CD
LYS
A
225
12.604
33.860
−10.427
1.00
52.08
A


ATOM
1700
CE
LYS
A
225
12.243
32.430
−10.017
1.00
53.90
A


ATOM
1701
NZ
LYS
A
225
10.779
32.155
−10.145
1.00
56.20
A


ATOM
1702
C
LYS
A
225
16.816
32.981
−9.734
1.00
38.89
A


ATOM
1703
O
LYS
A
225
16.693
31.812
−9.376
1.00
36.62
A


ATOM
1704
N
ILE
A
226
17.290
33.934
−8.939
1.00
37.52
A


ATOM
1705
CA
ILE
A
226
17.676
33.648
−7.565
1.00
35.91
A


ATOM
1706
CB
ILE
A
226
18.415
34.860
−6.939
1.00
34.66
A


ATOM
1707
CG2
ILE
A
226
18.474
34.725
−5.412
1.00
32.77
A


ATOM
1708
CG1
ILE
A
226
19.831
34.932
−7.526
1.00
35.67
A


ATOM
1709
CD1
ILE
A
226
20.616
36.173
−7.142
1.00
35.94
A


ATOM
1710
C
ILE
A
226
16.419
33.297
−6.775
1.00
35.16
A


ATOM
1711
O
ILE
A
226
15.420
34.010
−6.838
1.00
36.27
A


ATOM
1712
N
ASP
A
227
16.472
32.183
−6.048
1.00
34.90
A


ATOM
1713
CA
ASP
A
227
15.337
31.700
−5.260
1.00
35.04
A


ATOM
1714
CB
ASP
A
227
15.589
30.250
−4.845
1.00
37.48
A


ATOM
1715
CG
ASP
A
227
15.768
29.329
−6.041
1.00
40.26
A


ATOM
1716
OD1
ASP
A
227
14.795
29.161
−6.811
1.00
40.93
A


ATOM
1717
OD2
ASP
A
227
16.881
28.784
−6.213
1.00
40.47
A


ATOM
1718
C
ASP
A
227
15.009
32.536
−4.024
1.00
35.52
A


ATOM
1719
O
ASP
A
227
13.836
32.772
−3.710
1.00
33.67
A


ATOM
1720
N
ARG
A
228
16.045
32.966
−3.311
1.00
33.10
A


ATOM
1721
CA
ARG
A
228
15.859
33.774
−2.117
1.00
31.73
A


ATOM
1722
CB
ARG
A
228
15.297
32.910
−0.974
1.00
31.32
A


ATOM
1723
CG
ARG
A
228
16.153
31.704
−0.621
1.00
29.51
A


ATOM
1724
CD
ARG
A
228
15.468
30.793
0.405
1.00
32.46
A


ATOM
1725
NE
ARG
A
228
16.370
29.723
0.826
1.00
31.71
A


ATOM
1726
CZ
ARG
A
228
16.486
28.550
0.215
1.00
33.70
A


ATOM
1727
NH1
ARG
A
228
15.741
28.266
−0.848
1.00
35.15
A


ATOM
1728
NH2
ARG
A
228
17.382
27.675
0.646
1.00
35.72
A


ATOM
1729
C
ARG
A
228
17.189
34.408
−1.723
1.00
30.83
A


ATOM
1730
O
ARG
A
228
18.239
34.052
−2.264
1.00
32.29
A


ATOM
1731
N
LEU
A
229
17.134
35.353
−0.793
1.00
31.06
A


ATOM
1732
CA
LEU
A
229
18.327
36.066
−0.330
1.00
29.02
A


ATOM
1733
CB
LEU
A
229
18.365
37.451
−0.964
1.00
27.74
A


ATOM
1734
CG
LEU
A
229
19.548
38.344
−0.595
1.00
26.58
A


ATOM
1735
CD1
LEU
A
229
20.822
37.775
−1.210
1.00
24.82
A


ATOM
1736
CD2
LEU
A
229
19.266
39.765
−1.083
1.00
26.37
A


ATOM
1737
C
LEU
A
229
18.293
36.211
1.187
1.00
27.73
A


ATOM
1738
O
LEU
A
229
17.310
36.701
1.743
1.00
28.78
A


ATOM
1739
N
CYS
A
230
19.367
35.808
1.855
1.00
28.33
A


ATOM
1740
CA
CYS
A
230
19.404
35.894
3.313
1.00
29.29
A


ATOM
1741
CB
CYS
A
230
19.510
34.485
3.909
1.00
29.74
A


ATOM
1742
SG
CYS
A
230
19.579
34.453
5.743
1.00
31.41
A


ATOM
1743
C
CYS
A
230
20.523
36.777
3.878
1.00
28.54
A


ATOM
1744
O
CYS
A
230
21.661
36.749
3.406
1.00
28.22
A


ATOM
1745
N
PHE
A
231
20.172
37.574
4.882
1.00
28.82
A


ATOM
1746
CA
PHE
A
231
21.114
38.451
5.564
1.00
29.99
A


ATOM
1747
CB
PHE
A
231
20.539
39.867
5.695
1.00
28.76
A


ATOM
1748
CG
PHE
A
231
20.349
40.570
4.371
1.00
31.79
A


ATOM
1749
CD1
PHE
A
231
19.261
40.265
3.550
1.00
31.62
A


ATOM
1750
CD2
PHE
A
231
21.279
41.515
3.930
1.00
31.11
A


ATOM
1751
CE1
PHE
A
231
19.103
40.891
2.305
1.00
30.59
A


ATOM
1752
CE2
PHE
A
231
21.130
42.147
2.683
1.00
31.97
A


ATOM
1753
CZ
PHE
A
231
20.038
41.830
1.873
1.00
29.20
A


ATOM
1754
C
PHE
A
231
21.339
37.846
6.946
1.00
30.94
A


ATOM
1755
O
PHE
A
231
20.380
37.553
7.660
1.00
30.10
A


ATOM
1756
N
ALA
A
232
22.600
37.645
7.319
1.00
30.85
A


ATOM
1757
CA
ALA
A
232
22.916
37.051
8.617
1.00
31.42
A


ATOM
1758
CB
ALA
A
232
23.814
35.825
8.420
1.00
30.46
A


ATOM
1759
C
ALA
A
232
23.584
38.039
9.580
1.00
30.98
A


ATOM
1760
O
ALA
A
232
24.402
38.857
9.175
1.00
31.61
A


ATOM
1761
N
VAL
A
233
23.227
37.933
10.856
1.00
32.02
A


ATOM
1762
CA
VAL
A
233
23.757
38.794
11.911
1.00
33.05
A


ATOM
1763
CB
VAL
A
233
22.651
39.748
12.449
1.00
34.08
A


ATOM
1764
CG1
VAL
A
233
23.109
40.432
13.753
1.00
35.86
A


ATOM
1765
CG2
VAL
A
233
22.325
40.800
11.396
1.00
31.79
A


ATOM
1766
C
VAL
A
233
24.271
37.931
13.065
1.00
33.07
A


ATOM
1767
O
VAL
A
233
23.580
37.024
13.524
1.00
34.33
A


ATOM
1768
N
ILE
A
234
25.485
38.211
13.526
1.00
34.06
A


ATOM
1769
CA
ILE
A
234
26.073
37.449
14.623
1.00
36.42
A


ATOM
1770
CB
ILE
A
234
27.519
37.004
14.260
1.00
34.49
A


ATOM
1771
CG2
ILE
A
234
28.420
38.209
14.104
1.00
35.27
A


ATOM
1772
CG1
ILE
A
234
28.051
36.032
15.316
1.00
35.08
A


ATOM
1773
CD1
ILE
A
234
29.314
35.277
14.888
1.00
32.27
A


ATOM
1774
C
ILE
A
234
26.053
38.313
15.888
1.00
35.82
A


ATOM
1775
O
ILE
A
234
26.473
39.461
15.863
1.00
36.21
A


ATOM
1776
N
SER
A
235
25.549
37.772
16.989
1.00
38.61
A


ATOM
1777
CA
SER
A
235
25.471
38.561
18.219
1.00
40.77
A


ATOM
1778
CB
SER
A
235
24.385
39.633
18.065
1.00
41.97
A


ATOM
1779
OG
SER
A
235
24.220
40.392
19.248
1.00
43.56
A


ATOM
1780
C
SER
A
235
25.175
37.742
19.467
1.00
42.35
A


ATOM
1781
O
SER
A
235
24.793
36.574
19.384
1.00
41.79
A


ATOM
1782
N
ASN
A
236
25.356
38.372
20.626
1.00
44.38
A


ATOM
1783
CA
ASN
A
236
25.081
37.731
21.906
1.00
45.74
A


ATOM
1784
CB
ASN
A
236
26.115
38.151
22.958
1.00
47.60
A


ATOM
1785
CG
ASN
A
236
27.488
37.569
22.694
1.00
48.75
A


ATOM
1786
OD1
ASN
A
236
27.661
36.350
22.680
1.00
51.03
A


ATOM
1787
ND2
ASN
A
236
28.472
38.436
22.479
1.00
49.64
A


ATOM
1788
C
ASN
A
236
23.692
38.172
22.350
1.00
46.12
A


ATOM
1789
O
ASN
A
236
23.088
37.568
23.236
1.00
47.30
A


ATOM
1790
N
ASP
A
237
23.201
39.230
21.713
1.00
46.49
A


ATOM
1791
CA
ASP
A
237
21.889
39.806
21.993
1.00
46.83
A


ATOM
1792
CB
ASP
A
237
21.746
41.105
21.198
1.00
47.75
A


ATOM
1793
CG
ASP
A
237
20.662
42.013
21.736
1.00
47.79
A


ATOM
1794
OD1
ASP
A
237
19.513
41.560
21.923
1.00
47.65
A


ATOM
1795
OD2
ASP
A
237
20.970
43.198
21.957
1.00
48.75
A


ATOM
1796
C
ASP
A
237
20.786
38.825
21.591
1.00
47.10
A


ATOM
1797
O
ASP
A
237
20.665
38.463
20.422
1.00
47.73
A


ATOM
1798
N
PRO
A
238
19.965
38.383
22.557
1.00
46.57
A


ATOM
1799
CD
PRO
A
238
20.211
38.488
24.009
1.00
47.58
A


ATOM
1800
CA
PRO
A
238
18.878
37.439
22.281
1.00
45.52
A


ATOM
1801
CB
PRO
A
238
18.796
36.641
23.576
1.00
47.17
A


ATOM
1802
CG
PRO
A
238
19.055
37.703
24.605
1.00
47.42
A


ATOM
1803
C
PRO
A
238
17.530
38.049
21.893
1.00
45.24
A


ATOM
1804
O
PRO
A
238
16.509
37.361
21.900
1.00
45.57
A


ATOM
1805
N
THR
A
239
17.521
39.331
21.545
1.00
45.54
A


ATOM
1806
CA
THR
A
239
16.280
40.001
21.153
1.00
45.56
A


ATOM
1807
CB
THR
A
239
15.920
41.117
22.162
1.00
45.90
A


ATOM
1808
OG1
THR
A
239
16.886
42.173
22.067
1.00
46.13
A


ATOM
1809
CG2
THR
A
239
15.921
40.574
23.588
1.00
46.71
A


ATOM
1810
C
THR
A
239
16.372
40.642
19.761
1.00
44.94
A


ATOM
1811
O
THR
A
239
15.657
41.600
19.473
1.00
44.48
A


ATOM
1812
N
LEU
A
240
17.226
40.106
18.892
1.00
45.21
A


ATOM
1813
CA
LEU
A
240
17.412
40.689
17.563
1.00
44.68
A


ATOM
1814
CB
LEU
A
240
18.887
40.606
17.167
1.00
44.71
A


ATOM
1815
CG
LEU
A
240
19.890
41.396
18.008
1.00
45.86
A


ATOM
1816
CD1
LEU
A
240
21.292
41.136
17.488
1.00
46.25
A


ATOM
1817
CD2
LEU
A
240
19.566
42.885
17.952
1.00
45.57
A


ATOM
1818
C
LEU
A
240
16.573
40.190
16.387
1.00
44.93
A


ATOM
1819
O
LEU
A
240
16.801
40.625
15.258
1.00
44.99
A


ATOM
1820
N
VAL
A
241
15.614
39.298
16.617
1.00
44.69
A


ATOM
1821
CA
VAL
A
241
14.809
38.806
15.504
1.00
44.20
A


ATOM
1822
CB
VAL
A
241
13.775
37.744
15.976
1.00
44.62
A


ATOM
1823
CG1
VAL
A
241
12.661
38.393
16.781
1.00
45.74
A


ATOM
1824
CG2
VAL
A
241
13.219
36.998
14.778
1.00
43.96
A


ATOM
1825
C
VAL
A
241
14.114
40.004
14.839
1.00
44.51
A


ATOM
1826
O
VAL
A
241
13.424
40.780
15.498
1.00
44.10
A


ATOM
1827
N
PRO
A
242
14.302
40.175
13.520
1.00
42.66
A


ATOM
1828
CD
PRO
A
242
15.176
39.385
12.635
1.00
41.53
A


ATOM
1829
CA
PRO
A
242
13.696
41.293
12.787
1.00
41.75
A


ATOM
1830
CB
PRO
A
242
14.584
41.403
11.557
1.00
41.67
A


ATOM
1831
CG
PRO
A
242
14.865
39.966
11.257
1.00
41.45
A


ATOM
1832
C
PRO
A
242
12.229
41.111
12.419
1.00
42.06
A


ATOM
1833
O
PRO
A
242
11.849
41.287
11.265
1.00
39.87
A


ATOM
1834
N
SER
A
243
11.409
40.772
13.407
1.00
42.14
A


ATOM
1835
CA
SER
A
243
9.987
40.566
13.181
1.00
43.50
A


ATOM
1836
CB
SER
A
243
9.629
39.096
13.421
1.00
42.47
A


ATOM
1837
OG
SER
A
243
8.237
38.875
13.288
1.00
41.75
A


ATOM
1838
C
SER
A
243
9.170
41.460
14.109
1.00
45.15
A


ATOM
1839
O
SER
A
243
9.611
41.799
15.208
1.00
42.95
A


ATOM
1840
N
SER
A
244
7.981
41.844
13.657
1.00
47.40
A


ATOM
1841
CA
SER
A
244
7.102
42.697
14.450
1.00
49.90
A


ATOM
1842
CB
SER
A
244
6.520
43.813
13.579
1.00
49.67
A


ATOM
1843
OG
SER
A
244
5.769
43.277
12.500
1.00
48.66
A


ATOM
1844
C
SER
A
244
5.971
41.865
15.033
1.00
51.74
A


ATOM
1845
O
SER
A
244
5.122
42.375
15.765
1.00
53.20
A


ATOM
1846
N
ASP
A
245
5.964
40.580
14.693
1.00
53.53
A


ATOM
1847
CA
ASP
A
245
4.946
39.653
15.171
1.00
55.50
A


ATOM
1848
CB
ASP
A
245
4.823
38.470
14.207
1.00
56.42
A


ATOM
1849
CG
ASP
A
245
3.803
37.450
14.665
1.00
57.31
A


ATOM
1850
OD1
ASP
A
245
2.631
37.830
14.862
1.00
59.97
A


ATOM
1851
OD2
ASP
A
245
4.170
36.269
14.827
1.00
57.82
A


ATOM
1852
C
ASP
A
245
5.283
39.149
16.571
1.00
55.95
A


ATOM
1853
O
ASP
A
245
6.250
38.411
16.758
1.00
55.39
A


ATOM
1854
N
GLU
A
246
4.479
39.562
17.549
1.00
57.53
A


ATOM
1855
CA
GLU
A
246
4.673
39.162
18.941
1.00
58.27
A


ATOM
1856
CB
GLU
A
246
3.443
39.537
19.776
1.00
60.44
A


ATOM
1857
CG
GLU
A
246
3.265
41.033
19.999
1.00
63.33
A


ATOM
1858
CD
GLU
A
246
2.015
41.367
20.806
1.00
65.68
A


ATOM
1859
OE1
GLU
A
246
0.893
41.126
20.306
1.00
65.94
A


ATOM
1860
OE2
GLU
A
246
2.156
41.868
21.944
1.00
67.06
A


ATOM
1861
C
GLU
A
246
4.949
37.666
19.072
1.00
57.75
A


ATOM
1862
O
GLU
A
246
5.645
37.236
19.989
1.00
57.64
A


ATOM
1863
N
GLY
A
247
4.400
36.876
18.155
1.00
57.63
A


ATOM
1864
CA
GLY
A
247
4.625
35.440
18.196
1.00
57.31
A


ATOM
1865
C
GLY
A
247
6.097
35.098
18.032
1.00
56.48
A


ATOM
1866
O
GLY
A
247
6.657
34.325
18.814
1.00
56.42
A


ATOM
1867
N
ASP
A
248
6.731
35.677
17.016
1.00
55.47
A


ATOM
1868
CA
ASP
A
248
8.146
35.430
16.760
1.00
54.34
A


ATOM
1869
CB
ASP
A
248
8.576
36.044
15.426
1.00
52.59
A


ATOM
1870
CG
ASP
A
248
7.876
35.427
14.246
1.00
51.48
A


ATOM
1871
OD1
ASP
A
248
7.731
34.188
14.225
1.00
51.37
A


ATOM
1872
OD2
ASP
A
248
7.485
36.180
13.328
1.00
51.45
A


ATOM
1873
C
ASP
A
248
9.018
36.022
17.855
1.00
54.31
A


ATOM
1874
O
ASP
A
248
9.865
35.339
18.428
1.00
54.40
A


ATOM
1875
N
ILE
A
249
8.813
37.306
18.123
1.00
55.01
A


ATOM
1876
CA
ILE
A
249
9.578
38.028
19.131
1.00
55.17
A


ATOM
1877
CB
ILE
A
249
8.906
39.378
19.454
1.00
55.80
A


ATOM
1878
CG2
ILE
A
249
9.831
40.234
20.307
1.00
55.95
A


ATOM
1879
CG1
ILE
A
249
8.575
40.109
18.151
1.00
56.11
A


ATOM
1880
CD1
ILE
A
249
7.749
41.366
18.338
1.00
57.14
A


ATOM
1881
C
ILE
A
249
9.682
37.201
20.404
1.00
55.78
A


ATOM
1882
O
ILE
A
249
10.697
37.234
21.102
1.00
55.95
A


ATOM
1883
N
GLU
A
250
8.626
36.450
20.690
1.00
55.60
A


ATOM
1884
CA
GLU
A
250
8.577
35.611
21.875
1.00
55.90
A


ATOM
1885
CB
GLU
A
250
7.126
35.267
22.214
1.00
57.52
A


ATOM
1886
CG
GLU
A
250
6.987
34.358
23.416
1.00
60.65
A


ATOM
1887
CD
GLU
A
250
7.686
34.919
24.634
1.00
62.45
A


ATOM
1888
OE1
GLU
A
250
7.352
36.054
25.038
1.00
63.26
A


ATOM
1889
OE2
GLU
A
250
8.572
34.229
25.182
1.00
63.88
A


ATOM
1890
C
GLU
A
250
9.368
34.323
21.698
1.00
54.72
A


ATOM
1891
O
GLU
A
250
10.349
34.080
22.405
1.00
53.29
A


ATOM
1892
N
LYS
A
251
8.928
33.506
20.745
1.00
54.03
A


ATOM
1893
CA
LYS
A
251
9.555
32.221
20.461
1.00
53.48
A


ATOM
1894
CB
LYS
A
251
8.880
31.573
19.251
1.00
54.33
A


ATOM
1895
CG
LYS
A
251
9.268
30.122
19.033
1.00
55.38
A


ATOM
1896
CD
LYS
A
251
8.414
29.489
17.953
1.00
57.26
A


ATOM
1897
CE
LYS
A
251
8.646
27.989
17.873
1.00
58.08
A


ATOM
1898
NZ
LYS
A
251
7.790
27.348
16.833
1.00
58.69
A


ATOM
1899
C
LYS
A
251
11.067
32.292
20.235
1.00
52.15
A


ATOM
1900
O
LYS
A
251
11.812
31.441
20.731
1.00
51.69
A


ATOM
1901
N
PHE
A
252
11.523
33.300
19.497
1.00
50.10
A


ATOM
1902
CA
PHE
A
252
12.951
33.434
19.232
1.00
48.89
A


ATOM
1903
CB
PHE
A
252
13.197
34.435
18.099
1.00
45.87
A


ATOM
1904
CG
PHE
A
252
12.915
33.875
16.733
1.00
43.99
A


ATOM
1905
CD1
PHE
A
252
11.616
33.564
16.349
1.00
41.90
A


ATOM
1906
CD2
PHE
A
252
13.957
33.623
15.841
1.00
42.23
A


ATOM
1907
CE1
PHE
A
252
11.356
33.009
15.097
1.00
41.89
A


ATOM
1908
CE2
PHE
A
252
13.708
33.069
14.587
1.00
41.06
A


ATOM
1909
CZ
PHE
A
252
12.409
32.762
14.214
1.00
41.75
A


ATOM
1910
C
PHE
A
252
13.744
33.837
20.466
1.00
48.68
A


ATOM
1911
O
PHE
A
252
14.855
33.355
20.680
1.00
49.06
A


ATOM
1912
N
HIS
A
253
13.181
34.722
21.279
1.00
48.76
A


ATOM
1913
CA
HIS
A
253
13.870
35.145
22.490
1.00
49.30
A


ATOM
1914
CB
HIS
A
253
13.079
36.237
23.215
1.00
49.55
A


ATOM
1915
CG
HIS
A
253
13.576
36.513
24.599
1.00
50.62
A


ATOM
1916
CD2
HIS
A
253
14.564
37.324
25.045
1.00
50.93
A


ATOM
1917
ND1
HIS
A
253
13.084
35.861
25.709
1.00
51.60
A


ATOM
1918
CE1
HIS
A
253
13.749
36.258
26.779
1.00
52.53
A


ATOM
1919
NE2
HIS
A
253
14.653
37.144
26.404
1.00
52.16
A


ATOM
1920
C
HIS
A
253
14.033
33.937
23.403
1.00
48.72
A


ATOM
1921
O
HIS
A
253
15.067
33.765
24.052
1.00
47.39
A


ATOM
1922
N
ASN
A
254
12.995
33.108
23.437
1.00
48.76
A


ATOM
1923
CA
ASN
A
254
12.986
31.900
24.246
1.00
50.80
A


ATOM
1924
CB
ASN
A
254
11.657
31.165
24.060
1.00
52.29
A


ATOM
1925
CG
ASN
A
254
11.531
29.948
24.955
1.00
55.40
A


ATOM
1926
OD1
ASN
A
254
11.461
30.066
26.181
1.00
57.62
A


ATOM
1927
ND2
ASN
A
254
11.503
28.768
24.345
1.00
55.81
A


ATOM
1928
C
ASN
A
254
14.137
30.998
23.817
1.00
50.81
A


ATOM
1929
O
ASN
A
254
15.008
30.646
24.623
1.00
50.82
A


ATOM
1930
N
TYR
A
255
14.138
30.637
22.537
1.00
50.64
A


ATOM
1931
CA
TYR
A
255
15.171
29.773
21.977
1.00
49.68
A


ATOM
1932
CB
TYR
A
255
14.963
29.601
20.471
1.00
50.04
A


ATOM
1933
CG
TYR
A
255
15.905
28.596
19.842
1.00
50.36
A


ATOM
1934
CD1
TYR
A
255
15.670
27.228
19.959
1.00
49.51
A


ATOM
1935
CE1
TYR
A
255
16.550
26.296
19.415
1.00
49.32
A


ATOM
1936
CD2
TYR
A
255
17.047
29.012
19.162
1.00
49.32
A


ATOM
1937
CE2
TYR
A
255
17.936
28.088
18.614
1.00
49.59
A


ATOM
1938
CZ
TYR
A
255
17.680
26.732
18.746
1.00
48.94
A


ATOM
1939
OH
TYR
A
255
18.557
25.811
18.223
1.00
48.35
A


ATOM
1940
C
TYR
A
255
16.545
30.365
22.217
1.00
48.91
A


ATOM
1941
O
TYR
A
255
17.471
29.660
22.607
1.00
49.09
A


ATOM
1942
N
ALA
A
256
16.659
31.670
21.989
1.00
48.96
A


ATOM
1943
CA
ALA
A
256
17.918
32.392
22.145
1.00
49.88
A


ATOM
1944
CB
ALA
A
256
17.756
33.820
21.618
1.00
48.87
A


ATOM
1945
C
ALA
A
256
18.500
32.422
23.564
1.00
50.82
A


ATOM
1946
O
ALA
A
256
19.716
32.536
23.735
1.00
50.22
A


ATOM
1947
N
THR
A
257
17.651
32.328
24.582
1.00
52.17
A


ATOM
1948
CA
THR
A
257
18.152
32.352
25.954
1.00
53.69
A


ATOM
1949
CB
THR
A
257
17.347
33.334
26.837
1.00
52.46
A


ATOM
1950
OG1
THR
A
257
15.945
33.092
26.677
1.00
51.52
A


ATOM
1951
CG2
THR
A
257
17.667
34.771
26.462
1.00
52.76
A


ATOM
1952
C
THR
A
257
18.145
30.977
26.615
1.00
55.15
A


ATOM
1953
O
THR
A
257
18.703
30.804
27.699
1.00
55.75
A


ATOM
1954
N
LYS
A
258
17.528
29.997
25.959
1.00
56.37
A


ATOM
1955
CA
LYS
A
258
17.461
28.648
26.514
1.00
56.73
A


ATOM
1956
CB
LYS
A
258
16.001
28.247
26.728
1.00
58.42
A


ATOM
1957
CG
LYS
A
258
15.285
29.088
27.773
1.00
59.53
A


ATOM
1958
CD
LYS
A
258
13.861
28.613
27.997
1.00
60.71
A


ATOM
1959
CE
LYS
A
258
13.205
29.387
29.127
1.00
61.23
A


ATOM
1960
NZ
LYS
A
258
11.785
28.986
29.313
1.00
62.40
A


ATOM
1961
C
LYS
A
258
18.160
27.590
25.666
1.00
57.04
A


ATOM
1962
O
LYS
A
258
18.072
26.398
25.963
1.00
56.70
A


ATOM
1963
N
ALA
A
259
18.860
28.020
24.621
1.00
55.90
A


ATOM
1964
CA
ALA
A
259
19.557
27.082
23.743
1.00
55.40
A


ATOM
1965
CB
ALA
A
259
19.969
27.781
22.450
1.00
54.73
A


ATOM
1966
C
ALA
A
259
20.780
26.456
24.409
1.00
55.07
A


ATOM
1967
O
ALA
A
259
21.536
27.126
25.110
1.00
54.82
A


ATOM
1968
N
PRO
A
260
20.988
25.150
24.196
1.00
55.21
A


ATOM
1969
CD
PRO
A
260
20.123
24.180
23.501
1.00
54.90
A


ATOM
1970
CA
PRO
A
260
22.139
24.480
24.799
1.00
55.01
A


ATOM
1971
CB
PRO
A
260
21.806
23.003
24.615
1.00
55.58
A


ATOM
1972
CG
PRO
A
260
21.050
22.999
23.328
1.00
55.22
A


ATOM
1973
C
PRO
A
260
23.455
24.860
24.138
1.00
54.81
A


ATOM
1974
O
PRO
A
260
23.525
25.030
22.923
1.00
55.25
A


ATOM
1975
N
TYR
A
261
24.493
25.011
24.952
1.00
54.39
A


ATOM
1976
CA
TYR
A
261
25.818
25.337
24.452
1.00
55.68
A


ATOM
1977
CB
TYR
A
261
25.992
26.853
24.287
1.00
55.85
A


ATOM
1978
CG
TYR
A
261
25.843
27.685
25.542
1.00
55.90
A


ATOM
1979
CD1
TYR
A
261
26.945
28.326
26.108
1.00
55.27
A


ATOM
1980
CE1
TYR
A
261
26.809
29.139
27.232
1.00
55.94
A


ATOM
1981
CD2
TYR
A
261
24.594
27.873
26.136
1.00
55.74
A


ATOM
1982
CE2
TYR
A
261
24.446
28.682
27.261
1.00
56.36
A


ATOM
1983
CZ
TYR
A
261
25.558
29.313
27.803
1.00
56.36
A


ATOM
1984
OH
TYR
A
261
25.416
30.118
28.911
1.00
57.53
A


ATOM
1985
C
TYR
A
261
26.851
24.761
25.408
1.00
57.21
A


ATOM
1986
O
TYR
A
261
26.639
24.738
26.623
1.00
57.77
A


ATOM
1987
N
ALA
A
262
27.958
24.279
24.855
1.00
57.89
A


ATOM
1988
CA
ALA
A
262
29.010
23.668
25.655
1.00
58.96
A


ATOM
1989
CB
ALA
A
262
29.794
22.674
24.800
1.00
58.74
A


ATOM
1990
C
ALA
A
262
29.963
24.668
26.296
1.00
59.61
A


ATOM
1991
O
ALA
A
262
30.337
24.515
27.460
1.00
60.65
A


ATOM
1992
N
TYR
A
263
30.352
25.687
25.539
1.00
59.26
A


ATOM
1993
CA
TYR
A
263
31.276
26.705
26.027
1.00
58.56
A


ATOM
1994
CB
TYR
A
263
31.688
27.627
24.872
1.00
57.62
A


ATOM
1995
CG
TYR
A
263
30.532
28.126
24.031
1.00
55.64
A


ATOM
1996
CD1
TYR
A
263
29.825
27.261
23.193
1.00
55.16
A


ATOM
1997
CE1
TYR
A
263
28.753
27.712
22.431
1.00
53.62
A


ATOM
1998
CD2
TYR
A
263
30.134
29.460
24.082
1.00
55.16
A


ATOM
1999
CE2
TYR
A
263
29.061
29.922
23.323
1.00
53.97
A


ATOM
2000
CZ
TYR
A
263
28.375
29.044
22.503
1.00
53.51
A


ATOM
2001
OH
TYR
A
263
27.296
29.494
21.778
1.00
51.12
A


ATOM
2002
C
TYR
A
263
30.762
27.547
27.202
1.00
58.82
A


ATOM
2003
O
TYR
A
263
30.686
28.773
27.109
1.00
58.03
A


ATOM
2004
N
VAL
A
264
30.419
26.889
28.307
1.00
59.23
A


ATOM
2005
CA
VAL
A
264
29.941
27.593
29.496
1.00
59.65
A


ATOM
2006
CB
VAL
A
264
29.576
26.607
30.634
1.00
60.55
A


ATOM
2007
CG1
VAL
A
264
29.253
27.377
31.911
1.00
61.12
A


ATOM
2008
CG2
VAL
A
264
28.384
25.754
30.220
1.00
60.60
A


ATOM
2009
C
VAL
A
264
31.034
28.539
29.990
1.00
59.17
A


ATOM
2010
O
VAL
A
264
32.211
28.175
30.033
1.00
58.78
A


ATOM
2011
N
GLY
A
265
30.636
29.751
30.359
1.00
58.41
A


ATOM
2012
CA
GLY
A
265
31.591
30.739
30.824
1.00
57.69
A


ATOM
2013
C
GLY
A
265
31.473
31.966
29.945
1.00
57.54
A


ATOM
2014
O
GLY
A
265
31.800
33.086
30.341
1.00
56.90
A


ATOM
2015
N
GLU
A
266
31.009
31.740
28.724
1.00
57.93
A


ATOM
2016
CA
GLU
A
266
30.811
32.820
27.777
1.00
58.46
A


ATOM
2017
CB
GLU
A
266
31.588
32.567
26.490
1.00
58.78
A


ATOM
2018
CG
GLU
A
266
33.066
32.327
26.702
1.00
59.25
A


ATOM
2019
CD
GLU
A
266
33.790
32.041
25.405
1.00
59.38
A


ATOM
2020
OE1
GLU
A
266
34.004
32.986
24.616
1.00
59.35
A


ATOM
2021
OE2
GLU
A
266
34.135
30.865
25.171
1.00
60.57
A


ATOM
2022
C
GLU
A
266
29.326
32.844
27.480
1.00
58.93
A


ATOM
2023
O
GLU
A
266
28.578
31.961
27.913
1.00
59.06
A


ATOM
2024
N
LYS
A
267
28.893
33.855
26.743
1.00
58.87
A


ATOM
2025
CA
LYS
A
267
27.489
33.963
26.403
1.00
58.23
A


ATOM
2026
CB
LYS
A
267
27.080
35.438
26.338
1.00
59.85
A


ATOM
2027
CG
LYS
A
267
27.061
36.127
27.703
1.00
61.48
A


ATOM
2028
CD
LYS
A
267
28.411
36.008
28.406
1.00
62.59
A


ATOM
2029
CE
LYS
A
267
28.296
36.229
29.907
1.00
62.62
A


ATOM
2030
NZ
LYS
A
267
29.556
35.847
30.602
1.00
60.90
A


ATOM
2031
C
LYS
A
267
27.256
33.268
25.073
1.00
56.21
A


ATOM
2032
O
LYS
A
267
28.154
33.213
24.230
1.00
55.89
A


ATOM
2033
N
ARG
A
268
26.057
32.717
24.903
1.00
54.39
A


ATOM
2034
CA
ARG
A
268
25.698
32.023
23.674
1.00
52.55
A


ATOM
2035
CB
ARG
A
268
24.194
31.750
23.605
1.00
51.64
A


ATOM
2036
CG
ARG
A
268
23.639
30.751
24.585
1.00
52.17
A


ATOM
2037
CD
ARG
A
268
22.164
30.533
24.291
1.00
53.30
A


ATOM
2038
NE
ARG
A
268
21.522
29.641
25.251
1.00
55.82
A


ATOM
2039
CZ
ARG
A
268
21.311
29.938
26.530
1.00
56.60
A


ATOM
2040
NH1
ARG
A
268
21.689
31.114
27.014
1.00
57.37
A


ATOM
2041
NH2
ARG
A
268
20.722
29.055
27.325
1.00
57.84
A


ATOM
2042
C
ARG
A
268
26.053
32.847
22.454
1.00
50.70
A


ATOM
2043
O
ARG
A
268
25.856
34.065
22.435
1.00
51.42
A


ATOM
2044
N
THR
A
269
26.585
32.176
21.441
1.00
50.18
A


ATOM
2045
CA
THR
A
269
26.902
32.828
20.183
1.00
47.44
A


ATOM
2046
CB
THR
A
269
28.116
32.200
19.498
1.00
48.13
A


ATOM
2047
OG1
THR
A
269
29.289
32.466
20.277
1.00
48.94
A


ATOM
2048
CG2
THR
A
269
28.297
32.784
18.096
1.00
47.82
A


ATOM
2049
C
THR
A
269
25.656
32.572
19.343
1.00
46.15
A


ATOM
2050
O
THR
A
269
25.227
31.430
19.188
1.00
46.11
A


ATOM
2051
N
LEU
A
270
25.063
33.641
18.830
1.00
44.47
A


ATOM
2052
CA
LEU
A
270
23.853
33.532
18.031
1.00
43.06
A


ATOM
2053
CB
LEU
A
270
22.713
34.265
18.740
1.00
43.60
A


ATOM
2054
CG
LEU
A
270
22.468
33.872
20.200
1.00
44.62
A


ATOM
2055
CD1
LEU
A
270
21.764
35.002
20.927
1.00
44.69
A


ATOM
2056
CD2
LEU
A
270
21.646
32.597
20.261
1.00
44.54
A


ATOM
2057
C
LEU
A
270
24.054
34.133
16.642
1.00
40.92
A


ATOM
2058
O
LEU
A
270
24.858
35.046
16.462
1.00
42.68
A


ATOM
2059
N
VAL
A
271
23.332
33.603
15.663
1.00
39.89
A


ATOM
2060
CA
VAL
A
271
23.394
34.120
14.298
1.00
38.73
A


ATOM
2061
CB
VAL
A
271
24.243
33.236
13.371
1.00
39.03
A


ATOM
2062
CG1
VAL
A
271
24.009
33.628
11.911
1.00
37.91
A


ATOM
2063
CG2
VAL
A
271
25.703
33.419
13.699
1.00
37.99
A


ATOM
2064
C
VAL
A
271
21.972
34.181
13.775
1.00
37.49
A


ATOM
2065
O
VAL
A
271
21.330
33.154
13.568
1.00
36.59
A


ATOM
2066
N
TYR
A
272
21.475
35.398
13.602
1.00
37.97
A


ATOM
2067
CA
TYR
A
272
20.121
35.605
13.113
1.00
36.60
A


ATOM
2068
CB
TYR
A
272
19.528
36.903
13.659
1.00
37.13
A


ATOM
2069
CG
TYR
A
272
19.312
36.932
15.152
1.00
40.37
A


ATOM
2070
CD1
TYR
A
272
20.387
37.056
16.033
1.00
40.17
A


ATOM
2071
CE1
TYR
A
272
20.178
37.136
17.413
1.00
42.29
A


ATOM
2072
CD2
TYR
A
272
18.021
36.879
15.685
1.00
41.07
A


ATOM
2073
CE2
TYR
A
272
17.805
36.956
17.056
1.00
41.54
A


ATOM
2074
CZ
TYR
A
272
18.883
37.086
17.913
1.00
41.60
A


ATOM
2075
OH
TYR
A
272
18.657
37.172
19.266
1.00
43.79
A


ATOM
2076
C
TYR
A
272
20.109
35.686
11.603
1.00
35.69
A


ATOM
2077
O
TYR
A
272
21.119
35.983
10.977
1.00
34.57
A


ATOM
2078
N
GLY
A
273
18.940
35.444
11.031
1.00
34.52
A


ATOM
2079
CA
GLY
A
273
18.815
35.508
9.596
1.00
34.19
A


ATOM
2080
C
GLY
A
273
17.487
36.075
9.159
1.00
32.96
A


ATOM
2081
O
GLY
A
273
16.435
35.786
9.747
1.00
33.22
A


ATOM
2082
N
LEU
A
274
17.548
36.915
8.136
1.00
31.58
A


ATOM
2083
CA
LEU
A
274
16.360
37.508
7.557
1.00
30.07
A


ATOM
2084
CB
LEU
A
274
16.417
39.029
7.606
1.00
30.58
A


ATOM
2085
CG
LEU
A
274
15.329
39.712
6.766
1.00
32.48
A


ATOM
2086
CD1
LEU
A
274
13.961
39.283
7.256
1.00
32.49
A


ATOM
2087
CD2
LEU
A
274
15.470
41.216
6.855
1.00
31.20
A


ATOM
2088
C
LEU
A
274
16.403
37.044
6.119
1.00
29.70
A


ATOM
2089
O
LEU
A
274
17.318
37.399
5.377
1.00
27.86
A


ATOM
2090
N
THR
A
275
15.429
36.231
5.737
1.00
28.56
A


ATOM
2091
CA
THR
A
275
15.384
35.704
4.390
1.00
30.21
A


ATOM
2092
CB
THR
A
275
15.249
34.174
4.397
1.00
30.89
A


ATOM
2093
OG1
THR
A
275
16.243
33.611
5.263
1.00
31.27
A


ATOM
2094
CG2
THR
A
275
15.466
33.617
2.985
1.00
32.12
A


ATOM
2095
C
THR
A
275
14.228
36.296
3.613
1.00
31.45
A


ATOM
2096
O
THR
A
275
13.095
36.367
4.106
1.00
30.04
A


ATOM
2097
N
LEU
A
276
14.534
36.707
2.389
1.00
30.98
A


ATOM
2098
CA
LEU
A
276
13.560
37.306
1.502
1.00
32.05
A


ATOM
2099
CB
LEU
A
276
14.029
38.692
1.063
1.00
32.02
A


ATOM
2100
CG
LEU
A
276
14.463
39.658
2.161
1.00
31.99
A


ATOM
2101
CD1
LEU
A
276
15.012
40.926
1.528
1.00
33.02
A


ATOM
2102
CD2
LEU
A
276
13.287
39.968
3.067
1.00
31.46
A


ATOM
2103
C
LEU
A
276
13.377
36.442
0.269
1.00
32.27
A


ATOM
2104
O
LEU
A
276
14.331
36.202
−0.471
1.00
31.75
A


ATOM
2105
N
SER
A
277
12.151
35.967
0.065
1.00
33.88
A


ATOM
2106
CA
SER
A
277
11.830
35.164
−1.101
1.00
34.99
A


ATOM
2107
CB
SER
A
277
11.166
33.848
−0.689
1.00
35.49
A


ATOM
2108
OG
SER
A
277
9.833
34.061
−0.264
1.00
37.40
A


ATOM
2109
C
SER
A
277
10.873
36.020
−1.931
1.00
36.36
A


ATOM
2110
O
SER
A
277
10.451
37.090
−1.493
1.00
34.72
A


ATOM
2111
N
PRO
A
278
10.514
35.563
−3.139
1.00
39.44
A


ATOM
2112
CD
PRO
A
278
10.974
34.375
−3.875
1.00
39.05
A


ATOM
2113
CA
PRO
A
278
9.606
36.369
−3.957
1.00
40.64
A


ATOM
2114
CB
PRO
A
278
9.433
35.521
−5.214
1.00
40.18
A


ATOM
2115
CG
PRO
A
278
10.752
34.792
−5.306
1.00
40.69
A


ATOM
2116
C
PRO
A
278
8.272
36.695
−3.300
1.00
42.09
A


ATOM
2117
O
PRO
A
278
7.765
37.807
−3.428
1.00
43.87
A


ATOM
2118
N
LYS
A
279
7.715
35.740
−2.570
1.00
44.32
A


ATOM
2119
CA
LYS
A
279
6.414
35.959
−1.964
1.00
45.01
A


ATOM
2120
CB
LYS
A
279
5.533
34.738
−2.224
1.00
46.55
A


ATOM
2121
CG
LYS
A
279
5.493
34.307
−3.693
1.00
48.65
A


ATOM
2122
CD
LYS
A
279
4.868
35.368
−4.594
1.00
49.82
A


ATOM
2123
CE
LYS
A
279
4.878
34.913
−6.054
1.00
50.84
A


ATOM
2124
NZ
LYS
A
279
4.280
35.916
−6.979
1.00
51.34
A


ATOM
2125
C
LYS
A
279
6.380
36.303
−0.483
1.00
44.70
A


ATOM
2126
O
LYS
A
279
5.385
36.834
0.001
1.00
45.30
A


ATOM
2127
N
GLU
A
280
7.457
36.029
0.240
1.00
43.74
A


ATOM
2128
CA
GLU
A
280
7.464
36.308
1.670
1.00
42.41
A


ATOM
2129
CB
GLU
A
280
6.956
35.086
2.439
1.00
45.17
A


ATOM
2130
CG
GLU
A
280
7.607
33.770
1.997
1.00
48.54
A


ATOM
2131
CD
GLU
A
280
7.531
32.665
3.051
1.00
51.12
A


ATOM
2132
OE1
GLU
A
280
6.653
32.737
3.940
1.00
53.02
A


ATOM
2133
OE2
GLU
A
280
8.346
31.717
2.982
1.00
50.28
A


ATOM
2134
C
GLU
A
280
8.816
36.688
2.244
1.00
40.99
A


ATOM
2135
O
GLU
A
280
9.796
36.885
1.527
1.00
37.74
A


ATOM
2136
N
GLU
A
281
8.828
36.773
3.569
1.00
40.26
A


ATOM
2137
CA
GLU
A
281
10.003
37.079
4.363
1.00
39.86
A


ATOM
2138
CB
GLU
A
281
10.008
38.562
4.754
1.00
40.81
A


ATOM
2139
CG
GLU
A
281
10.698
38.866
6.077
1.00
41.06
A


ATOM
2140
CD
GLU
A
281
10.540
40.316
6.517
1.00
43.02
A


ATOM
2141
OE1
GLU
A
281
10.990
41.219
5.784
1.00
42.38
A


ATOM
2142
OE2
GLU
A
281
9.967
40.558
7.603
1.00
42.59
A


ATOM
2143
C
GLU
A
281
9.874
36.224
5.621
1.00
39.25
A


ATOM
2144
O
GLU
A
281
8.813
36.204
6.242
1.00
39.38
A


ATOM
2145
N
TYR
A
282
10.929
35.501
5.984
1.00
36.51
A


ATOM
2146
CA
TYR
A
282
10.895
34.703
7.200
1.00
35.17
A


ATOM
2147
CB
TYR
A
282
10.575
33.231
6.909
1.00
34.96
A


ATOM
2148
CG
TYR
A
282
11.546
32.493
6.022
1.00
35.52
A


ATOM
2149
CD1
TYR
A
282
12.574
31.728
6.569
1.00
35.03
A


ATOM
2150
CE1
TYR
A
282
13.446
31.001
5.758
1.00
36.48
A


ATOM
2151
CD2
TYR
A
282
11.411
32.521
4.631
1.00
34.37
A


ATOM
2152
CE2
TYR
A
282
12.279
31.796
3.809
1.00
34.23
A


ATOM
2153
CZ
TYR
A
282
13.291
31.040
4.380
1.00
34.88
A


ATOM
2154
OH
TYR
A
282
14.143
30.310
3.584
1.00
37.85
A


ATOM
2155
C
TYR
A
282
12.205
34.871
7.956
1.00
34.80
A


ATOM
2156
O
TYR
A
282
13.182
35.398
7.415
1.00
32.60
A


ATOM
2157
N
TYR
A
283
12.216
34.423
9.206
1.00
34.19
A


ATOM
2158
CA
TYR
A
283
13.369
34.598
10.080
1.00
33.80
A


ATOM
2159
CB
TYR
A
283
12.910
35.412
11.288
1.00
35.23
A


ATOM
2160
CG
TYR
A
283
11.860
36.424
10.889
1.00
36.91
A


ATOM
2161
CD1
TYR
A
283
12.193
37.541
10.117
1.00
37.31
A


ATOM
2162
CE1
TYR
A
283
11.206
38.407
9.629
1.00
37.64
A


ATOM
2163
CD2
TYR
A
283
10.512
36.203
11.175
1.00
38.08
A


ATOM
2164
CE2
TYR
A
283
9.521
37.059
10.695
1.00
38.79
A


ATOM
2165
CZ
TYR
A
283
9.875
38.154
9.923
1.00
39.16
A


ATOM
2166
OH
TYR
A
283
8.890
38.986
9.446
1.00
39.66
A


ATOM
2167
C
TYR
A
283
14.041
33.302
10.519
1.00
33.97
A


ATOM
2168
O
TYR
A
283
13.404
32.258
10.627
1.00
34.03
A


ATOM
2169
N
LYS
A
284
15.342
33.375
10.768
1.00
32.82
A


ATOM
2170
CA
LYS
A
284
16.086
32.191
11.170
1.00
33.38
A


ATOM
2171
CB
LYS
A
284
16.893
31.668
9.974
1.00
34.19
A


ATOM
2172
CG
LYS
A
284
15.989
31.260
8.809
1.00
36.80
A


ATOM
2173
CD
LYS
A
284
16.733
30.629
7.633
1.00
39.12
A


ATOM
2174
CE
LYS
A
284
17.536
31.643
6.843
1.00
40.11
A


ATOM
2175
NZ
LYS
A
284
18.047
31.069
5.548
1.00
34.37
A


ATOM
2176
C
LYS
A
284
16.988
32.481
12.362
1.00
32.66
A


ATOM
2177
O
LYS
A
284
17.392
33.624
12.583
1.00
33.84
A


ATOM
2178
N
LEU
A
285
17.283
31.448
13.143
1.00
33.01
A


ATOM
2179
CA
LEU
A
285
18.141
31.621
14.308
1.00
33.94
A


ATOM
2180
CB
LEU
A
285
17.309
32.021
15.531
1.00
34.70
A


ATOM
2181
CG
LEU
A
285
18.063
32.102
16.862
1.00
36.10
A


ATOM
2182
CD1
LEU
A
285
19.214
33.085
16.752
1.00
35.16
A


ATOM
2183
CD2
LEU
A
285
17.097
32.525
17.965
1.00
36.19
A


ATOM
2184
C
LEU
A
285
18.950
30.377
14.633
1.00
33.22
A


ATOM
2185
O
LEU
A
285
18.401
29.311
14.918
1.00
34.61
A


ATOM
2186
N
GLY
A
286
20.265
30.527
14.582
1.00
33.99
A


ATOM
2187
CA
GLY
A
286
21.142
29.422
14.893
1.00
34.28
A


ATOM
2188
C
GLY
A
286
21.824
29.682
16.218
1.00
34.79
A


ATOM
2189
O
GLY
A
286
22.402
30.745
16.418
1.00
35.42
A


ATOM
2190
N
ALA
A
287
21.728
28.726
17.135
1.00
36.63
A


ATOM
2191
CA
ALA
A
287
22.369
28.848
18.439
1.00
37.80
A


ATOM
2192
CB
ALA
A
287
21.384
28.517
19.546
1.00
37.09
A


ATOM
2193
C
ALA
A
287
23.530
27.865
18.454
1.00
38.30
A


ATOM
2194
O
ALA
A
287
23.326
26.648
18.412
1.00
38.88
A


ATOM
2195
N
TYR
A
288
24.746
28.397
18.497
1.00
40.30
A


ATOM
2196
CA
TYR
A
288
25.936
27.569
18.505
1.00
42.62
A


ATOM
2197
CB
TYR
A
288
27.189
28.434
18.343
1.00
43.09
A


ATOM
2198
CG
TYR
A
288
27.362
29.009
16.955
1.00
43.83
A


ATOM
2199
CD1
TYR
A
288
26.405
29.865
16.409
1.00
45.17
A


ATOM
2200
CE1
TYR
A
288
26.545
30.373
15.119
1.00
44.97
A


ATOM
2201
CD2
TYR
A
288
28.469
28.679
16.175
1.00
43.95
A


ATOM
2202
CE2
TYR
A
288
28.621
29.182
14.885
1.00
43.70
A


ATOM
2203
CZ
TYR
A
288
27.654
30.027
14.365
1.00
45.56
A


ATOM
2204
OH
TYR
A
288
27.789
30.525
13.090
1.00
46.87
A


ATOM
2205
C
TYR
A
288
26.059
26.734
19.768
1.00
45.18
A


ATOM
2206
O
TYR
A
288
25.919
27.240
20.882
1.00
44.32
A


ATOM
2207
N
TYR
A
289
26.305
25.443
19.573
1.00
46.25
A


ATOM
2208
CA
TYR
A
289
26.496
24.519
20.674
1.00
47.59
A


ATOM
2209
CB
TYR
A
289
25.997
23.126
20.305
1.00
48.44
A


ATOM
2210
CG
TYR
A
289
26.224
22.105
21.393
1.00
51.30
A


ATOM
2211
CD1
TYR
A
289
25.700
22.294
22.674
1.00
51.82
A


ATOM
2212
CE1
TYR
A
289
25.937
21.378
23.688
1.00
53.60
A


ATOM
2213
CD2
TYR
A
289
26.988
20.965
21.155
1.00
50.81
A


ATOM
2214
CE2
TYR
A
289
27.230
20.041
22.162
1.00
53.45
A


ATOM
2215
CZ
TYR
A
289
26.702
20.255
23.426
1.00
53.78
A


ATOM
2216
OH
TYR
A
289
26.946
19.346
24.428
1.00
55.63
A


ATOM
2217
C
TYR
A
289
27.998
24.496
20.912
1.00
47.53
A


ATOM
2218
O
TYR
A
289
28.458
24.476
22.053
1.00
48.27
A


ATOM
2219
N
HIS
A
290
28.755
24.505
19.819
1.00
46.65
A


ATOM
2220
CA
HIS
A
290
30.214
24.524
19.871
1.00
45.99
A


ATOM
2221
CB
HIS
A
290
30.824
23.263
19.246
1.00
47.09
A


ATOM
2222
CG
HIS
A
290
30.663
22.026
20.070
1.00
47.66
A


ATOM
2223
CD2
HIS
A
290
30.004
21.803
21.231
1.00
48.23
A


ATOM
2224
ND1
HIS
A
290
31.203
20.815
19.697
1.00
48.48
A


ATOM
2225
CE1
HIS
A
290
30.880
19.897
20.592
1.00
48.01
A


ATOM
2226
NE2
HIS
A
290
30.152
20.471
21.532
1.00
47.65
A


ATOM
2227
C
HIS
A
290
30.695
25.715
19.061
1.00
46.29
A


ATOM
2228
O
HIS
A
290
30.152
26.000
17.986
1.00
45.19
A


ATOM
2229
N
ILE
A
291
31.708
26.407
19.573
1.00
44.93
A


ATOM
2230
CA
ILE
A
291
32.282
27.548
18.875
1.00
45.16
A


ATOM
2231
CB
ILE
A
291
31.889
28.907
19.524
1.00
45.11
A


ATOM
2232
CG2
ILE
A
291
30.417
29.188
19.301
1.00
43.33
A


ATOM
2233
CG1
ILE
A
291
32.243
28.899
21.019
1.00
45.59
A


ATOM
2234
CD1
ILE
A
291
32.186
30.259
21.675
1.00
45.30
A


ATOM
2235
C
ILE
A
291
33.798
27.454
18.879
1.00
45.61
A


ATOM
2236
O
ILE
A
291
34.387
26.651
19.602
1.00
47.34
A


ATOM
2237
N
THR
A
292
34.424
28.291
18.065
1.00
47.17
A


ATOM
2238
CA
THR
A
292
35.875
28.341
17.967
1.00
47.28
A


ATOM
2239
CB
THR
A
292
36.371
27.755
16.639
1.00
47.48
A


ATOM
2240
OG1
THR
A
292
35.993
28.617
15.555
1.00
44.68
A


ATOM
2241
CG2
THR
A
292
35.775
26.374
16.426
1.00
46.70
A


ATOM
2242
C
THR
A
292
36.212
29.814
18.001
1.00
47.83
A


ATOM
2243
O
THR
A
292
35.307
30.644
18.064
1.00
48.07
A


ATOM
2244
N
ASP
A
293
37.497
30.149
17.954
1.00
48.59
A


ATOM
2245
CA
ASP
A
293
37.883
31.551
17.982
1.00
48.64
A


ATOM
2246
CB
ASP
A
293
39.408
31.705
17.988
1.00
49.60
A


ATOM
2247
CG
ASP
A
293
40.045
31.150
19.249
1.00
50.84
A


ATOM
2248
OD1
ASP
A
293
39.394
31.201
20.317
1.00
52.13
A


ATOM
2249
OD2
ASP
A
293
41.200
30.673
19.176
1.00
51.40
A


ATOM
2250
C
ASP
A
293
37.299
32.284
16.784
1.00
48.56
A


ATOM
2251
O
ASP
A
293
37.058
33.488
16.851
1.00
49.15
A


ATOM
2252
N
VAL
A
294
37.070
31.561
15.689
1.00
47.81
A


ATOM
2253
CA
VAL
A
294
36.512
32.179
14.487
1.00
46.87
A


ATOM
2254
CB
VAL
A
294
36.140
31.120
13.418
1.00
47.55
A


ATOM
2255
CG1
VAL
A
294
35.346
31.772
12.291
1.00
47.30
A


ATOM
2256
CG2
VAL
A
294
37.409
30.490
12.847
1.00
47.74
A


ATOM
2257
C
VAL
A
294
35.272
32.985
14.843
1.00
45.87
A


ATOM
2258
O
VAL
A
294
35.123
34.130
14.416
1.00
46.94
A


ATOM
2259
N
GLN
A
295
34.389
32.380
15.629
1.00
45.48
A


ATOM
2260
CA
GLN
A
295
33.162
33.035
16.061
1.00
45.44
A


ATOM
2261
CB
GLN
A
295
32.190
32.002
16.647
1.00
45.04
A


ATOM
2262
CG
GLN
A
295
31.550
31.080
15.603
1.00
44.79
A


ATOM
2263
CD
GLN
A
295
32.555
30.174
14.906
1.00
45.62
A


ATOM
2264
OE1
GLN
A
295
32.408
29.860
13.720
1.00
44.71
A


ATOM
2265
NE2
GLN
A
295
33.576
29.741
15.642
1.00
42.80
A


ATOM
2266
C
GLN
A
295
33.472
34.118
17.096
1.00
45.66
A


ATOM
2267
O
GLN
A
295
32.840
35.178
17.109
1.00
44.55
A


ATOM
2268
N
ARG
A
296
34.449
33.850
17.960
1.00
45.23
A


ATOM
2269
CA
ARG
A
296
34.842
34.819
18.982
1.00
45.03
A


ATOM
2270
CB
ARG
A
296
35.964
34.255
19.862
1.00
44.33
A


ATOM
2271
CG
ARG
A
296
35.592
32.952
20.540
1.00
44.26
A


ATOM
2272
CD
ARG
A
296
36.126
32.857
21.970
1.00
45.12
A


ATOM
2273
NE
ARG
A
296
35.665
31.624
22.597
1.00
44.32
A


ATOM
2274
CZ
ARG
A
296
36.124
30.419
22.280
1.00
44.21
A


ATOM
2275
NH1
ARG
A
296
37.069
30.292
21.357
1.00
44.51
A


ATOM
2276
NH2
ARG
A
296
35.613
29.341
22.860
1.00
45.05
A


ATOM
2277
C
ARG
A
296
35.317
36.091
18.297
1.00
44.39
A


ATOM
2278
O
ARG
A
296
35.067
37.202
18.766
1.00
44.29
A


ATOM
2279
N
GLY
A
297
36.004
35.917
17.175
1.00
43.83
A


ATOM
2280
CA
GLY
A
297
36.494
37.061
16.436
1.00
44.22
A


ATOM
2281
C
GLY
A
297
35.372
37.832
15.766
1.00
45.50
A


ATOM
2282
O
GLY
A
297
35.370
39.064
15.771
1.00
46.22
A


ATOM
2283
N
LEU
A
298
34.417
37.108
15.188
1.00
45.75
A


ATOM
2284
CA
LEU
A
298
33.292
37.732
14.498
1.00
46.53
A


ATOM
2285
CB
LEU
A
298
32.402
36.663
13.848
1.00
45.17
A


ATOM
2286
CG
LEU
A
298
33.021
35.931
12.650
1.00
45.06
A


ATOM
2287
CD1
LEU
A
298
32.084
34.824
12.164
1.00
44.48
A


ATOM
2288
CD2
LEU
A
298
33.307
36.932
11.537
1.00
45.31
A


ATOM
2289
C
LEU
A
298
32.462
38.602
15.433
1.00
47.62
A


ATOM
2290
O
LEU
A
298
31.945
39.646
15.028
1.00
47.53
A


ATOM
2291
N
LEU
A
299
32.340
38.171
16.685
1.00
48.72
A


ATOM
2292
CA
LEU
A
299
31.580
38.924
17.667
1.00
50.18
A


ATOM
2293
CB
LEU
A
299
31.390
38.090
18.930
1.00
49.60
A


ATOM
2294
CG
LEU
A
299
30.350
36.975
18.807
1.00
48.08
A


ATOM
2295
CD1
LEU
A
299
30.486
36.019
19.974
1.00
47.37
A


ATOM
2296
CD2
LEU
A
299
28.953
37.573
18.765
1.00
48.29
A


ATOM
2297
C
LEU
A
299
32.240
40.260
18.004
1.00
51.67
A


ATOM
2298
O
LEU
A
299
31.550
41.240
18.282
1.00
51.55
A


ATOM
2299
N
LYS
A
300
33.572
40.303
17.970
1.00
53.35
A


ATOM
2300
CA
LYS
A
300
34.304
41.537
18.263
1.00
53.76
A


ATOM
2301
CB
LYS
A
300
35.794
41.265
18.493
1.00
54.63
A


ATOM
2302
CG
LYS
A
300
36.147
40.439
19.703
1.00
56.78
A


ATOM
2303
CD
LYS
A
300
37.663
40.361
19.835
1.00
58.59
A


ATOM
2304
CE
LYS
A
300
38.091
39.473
20.991
1.00
60.33
A


ATOM
2305
NZ
LYS
A
300
39.558
39.580
21.244
1.00
61.89
A


ATOM
2306
C
LYS
A
300
34.192
42.492
17.086
1.00
53.47
A


ATOM
2307
O
LYS
A
300
34.072
43.705
17.259
1.00
54.79
A


ATOM
2308
N
ALA
A
301
34.241
41.933
15.885
1.00
51.85
A


ATOM
2309
CA
ALA
A
301
34.172
42.729
14.669
1.00
50.07
A


ATOM
2310
CB
ALA
A
301
34.698
41.913
13.495
1.00
50.60
A


ATOM
2311
C
ALA
A
301
32.791
43.272
14.332
1.00
49.04
A


ATOM
2312
O
ALA
A
301
32.672
44.404
13.855
1.00
48.99
A


ATOM
2313
N
PHE
A
302
31.750
42.479
14.583
1.00
47.91
A


ATOM
2314
CA
PHE
A
302
30.391
42.895
14.244
1.00
46.87
A


ATOM
2315
CB
PHE
A
302
29.775
41.920
13.229
1.00
45.69
A


ATOM
2316
CG
PHE
A
302
30.542
41.819
11.943
1.00
41.83
A


ATOM
2317
CD1
PHE
A
302
31.567
40.890
11.801
1.00
41.69
A


ATOM
2318
CD2
PHE
A
302
30.256
42.672
10.881
1.00
42.51
A


ATOM
2319
CE1
PHE
A
302
32.298
40.813
10.620
1.00
40.43
A


ATOM
2320
CE2
PHE
A
302
30.982
42.603
9.693
1.00
40.70
A


ATOM
2321
CZ
PHE
A
302
32.004
41.675
9.562
1.00
41.10
A


ATOM
2322
C
PHE
A
302
29.418
43.061
15.398
1.00
47.58
A


ATOM
2323
O
PHE
A
302
28.303
43.536
15.198
1.00
47.34
A


ATOM
2324
N
ASP
A
303
29.817
42.668
16.600
1.00
48.39
A


ATOM
2325
CA
ASP
A
303
28.919
42.807
17.738
1.00
49.75
A


ATOM
2326
CB
ASP
A
303
28.481
41.423
18.224
1.00
48.37
A


ATOM
2327
CG
ASP
A
303
27.185
41.462
18.998
1.00
47.54
A


ATOM
2328
OD1
ASP
A
303
26.215
42.066
18.497
1.00
47.88
A


ATOM
2329
OD2
ASP
A
303
27.126
40.879
20.100
1.00
49.46
A


ATOM
2330
C
ASP
A
303
29.581
43.589
18.874
1.00
50.31
A


ATOM
2331
O
ASP
A
303
29.661
43.050
19.995
1.00
51.55
A


ATOM
2332
OXT
ASP
A
303
30.012
44.737
18.627
1.00
51.61
A


ATOM
2333
MG+2
MG2
A
1
19.221
25.308
7.555
1.00
43.67
M


ATOM
2334
PA
GSP
A
1
19.534
27.534
3.619
1.00
40.67
G


ATOM
2335
O1A
GSP
A
1
19.609
28.204
2.288
1.00
39.03
G


ATOM
2336
O2A
GSP
A
1
20.362
26.298
3.674
1.00
39.43
G


ATOM
2337
O3A
GSP
A
1
18.121
27.257
4.002
1.00
42.12
G


ATOM
2338
O1B
GSP
A
1
20.124
28.585
4.679
1.00
40.27
G


ATOM
2339
PB
GSP
A
1
20.529
28.366
6.198
1.00
40.45
G


ATOM
2340
O2B
GSP
A
1
20.225
29.576
7.030
1.00
40.45
G


ATOM
2341
O3B
GSP
A
1
19.816
27.203
6.821
1.00
40.17
G


ATOM
2342
S1
GSP
A
1
22.118
28.106
6.368
1.00
46.86
G


ATOM
2343
C1
GSP
A
1
22.785
28.278
7.611
1.00
48.16
G


ATOM
2344
C2
GSP
A
1
23.948
27.320
7.744
1.00
48.30
G


ATOM
2345
C3
GSP
A
1
25.237
27.674
7.899
1.00
48.85
G


ATOM
2346
C10
GSP
A
1
25.862
28.840
7.172
1.00
48.34
G


ATOM
2347
C4
GSP
A
1
26.165
26.878
8.794
1.00
49.42
G


ATOM
2348
C5
GSP
A
1
27.559
27.525
8.868
1.00
51.23
G


ATOM
2349
C6
GSP
A
1
28.644
26.503
9.103
1.00
53.24
G


ATOM
2350
C7
GSP
A
1
29.414
26.374
10.200
1.00
53.60
G


ATOM
2351
C9
GSP
A
1
30.505
27.374
10.507
1.00
54.65
G


ATOM
2352
C8
GSP
A
1
29.296
25.257
11.211
1.00
55.35
G


ATOM
2353
C1
DH2
A
1
26.153
31.881
8.710
1.00
65.18
D


ATOM
2354
C2
DH2
A
1
24.959
31.390
9.301
1.00
65.39
D


ATOM
2355
C3
DH2
A
1
25.028
30.627
10.451
1.00
66.41
D


ATOM
2356
C4
DH2
A
1
26.279
30.324
11.055
1.00
66.26
D


ATOM
2357
C5
DH2
A
1
27.435
30.796
10.488
1.00
66.06
D


ATOM
2358
C6
DH2
A
1
28.585
32.099
8.675
1.00
64.51
D


ATOM
2359
C7
DH2
A
1
27.251
33.138
6.948
1.00
63.33
D


ATOM
2360
C8
DH2
A
1
26.099
32.663
7.525
1.00
63.43
D


ATOM
2361
O1
DH2
A
1
29.857
31.880
9.155
1.00
65.00
D


ATOM
2362
O2
DH2
A
1
23.839
30.179
10.983
1.00
66.99
D


ATOM
2363
C9
DH2
A
1
28.509
32.861
7.519
1.00
64.14
D


ATOM
2364
C10
DH2
A
1
27.409
31.584
9.306
1.00
65.41
D


ATOM
2365
N
NO3
A
1
42.060
34.777
−5.122
1.00
69.01
N


ATOM
2366
O1
NO3
A
1
41.197
34.277
−5.872
1.00
69.03
N


ATOM
2367
O2
NO3
A
1
41.725
35.701
−4.452
1.00
68.04
N


ATOM
2368
O3
NO3
A
1
43.135
34.230
−5.061
1.00
68.81
N


ATOM
2369
OH2
TIP
A
1
30.490
35.181
26.300
1.00
65.42
W


ATOM
2370
OH2
TIP
A
2
36.852
21.356
11.220
1.00
62.71
W


ATOM
2371
OH2
TIP
A
3
13.275
6.277
5.899
1.00
52.03
W


ATOM
2372
OH2
TIP
A
4
39.421
26.056
19.137
1.00
55.68
W


ATOM
2373
OH2
TIP
A
5
29.221
47.052
19.171
1.00
51.85
W


ATOM
2374
OH2
TIP
A
6
25.560
30.322
10.756
1.00
97.83
W


ATOM
2375
OH2
TIP
A
7
23.552
52.697
−2.797
1.00
48.74
W


ATOM
2376
OH2
TIP
A
8
15.783
28.067
4.579
1.00
42.75
W


ATOM
2377
OH2
TIP
A
9
35.855
25.568
−13.555
1.00
79.37
W


ATOM
2378
OH2
TIP
A
10
36.538
42.420
−0.112
1.00
49.25
W


ATOM
2379
OH2
TIP
A
11
23.745
35.145
24.150
1.00
53.80
W


ATOM
2380
OH2
TIP
A
12
19.673
51.064
6.990
1.00
47.69
W


ATOM
2381
OH2
TIP
A
13
27.368
30.522
28.673
1.00
65.03
W


ATOM
2382
OH2
TIP
A
14
32.186
14.701
−3.539
1.00
56.64
W


ATOM
2383
OH2
TIP
A
15
24.460
14.475
−13.361
1.00
59.07
W


ATOM
2384
OH2
TIP
A
16
40.585
17.057
1.471
1.00
59.15
W


ATOM
2385
OH2
TIP
A
17
17.913
13.365
−5.351
1.00
45.07
W


ATOM
2386
OH2
TIP
A
18
15.723
27.642
7.214
1.00
47.54
W


ATOM
2387
OH2
TIP
A
19
22.263
7.047
−6.225
1.00
59.29
W


ATOM
2388
OH2
TIP
A
20
19.969
29.141
−17.580
1.00
67.73
W


ATOM
2389
OH2
TIP
A
21
21.634
48.493
12.171
1.00
47.20
W


ATOM
2390
OH2
TIP
A
22
18.106
22.881
−16.847
1.00
50.83
W


ATOM
2391
OH2
TIP
A
23
29.495
40.536
21.854
1.00
60.58
W


ATOM
2392
OH2
TIP
A
24
36.302
27.268
13.132
1.00
55.92
W


ATOM
2393
OH2
TIP
A
25
29.732
35.121
4.882
1.00
50.52
W


ATOM
2394
OH2
TIP
A
26
45.475
29.946
0.240
1.00
44.34
W


ATOM
2395
OH2
TIP
A
27
15.795
10.968
19.889
1.00
77.54
W


ATOM
2396
OH2
TIP
A
28
14.431
30.709
−8.734
1.00
39.58
W


ATOM
2397
OH2
TIP
A
29
13.313
28.671
−1.901
1.00
44.66
W


ATOM
2398
OH2
TIP
A
30
27.440
52.388
8.029
1.00
60.67
W


ATOM
2399
OH2
TIP
A
31
42.559
22.415
4.064
1.00
53.62
W


ATOM
2400
OH2
TIP
A
32
20.941
43.226
24.809
1.00
55.10
W


ATOM
2401
OH2
TIP
A
33
33.359
44.497
−2.388
1.00
58.45
W


ATOM
2402
OH2
TIP
A
34
31.147
53.165
−0.332
1.00
46.23
W


ATOM
2403
OH2
TIP
A
35
12.356
25.536
1.697
1.00
64.15
W


ATOM
2404
OH2
TIP
A
36
35.138
10.636
12.652
1.00
61.46
W


ATOM
2405
OH2
TIP
A
37
15.020
50.521
−4.580
1.00
40.30
W


ATOM
2406
OH2
TIP
A
38
9.781
27.922
21.238
1.00
68.26
W


ATOM
2407
OH2
TIP
A
39
41.691
22.533
−2.565
1.00
54.13
W


ATOM
2408
OH2
TIP
A
40
20.666
45.859
15.289
1.00
70.16
W


ATOM
2409
OH2
TIP
A
41
34.363
37.353
21.374
1.00
53.19
W


ATOM
2410
OH2
TIP
A
42
26.734
5.871
−6.191
1.00
45.50
W


ATOM
2411
OH2
TIP
A
43
10.365
49.794
12.437
1.00
50.70
W


ATOM
2412
OH2
TIP
A
44
34.543
29.704
9.768
1.00
55.92
W


ATOM
2413
OH2
TIP
A
45
40.376
31.076
6.686
1.00
58.22
W


ATOM
2414
OH2
TIP
A
46
34.553
5.913
23.858
1.00
71.52
W


ATOM
2415
OH2
TIP
A
47
14.200
27.005
5.946
1.00
53.11
W


ATOM
2416
OH2
TIP
A
48
42.976
30.424
−5.522
1.00
44.67
W


ATOM
2417
OH2
TIP
A
49
31.686
41.102
21.600
1.00
49.60
W


ATOM
2418
OH2
TIP
A
50
37.985
29.637
6.203
1.00
60.32
W


ATOM
2419
OH2
TIP
A
51
16.521
36.553
−8.898
1.00
49.77
W


ATOM
2420
OH2
TIP
A
52
31.859
17.997
−13.734
1.00
53.29
W


ATOM
2421
OH2
TIP
A
53
27.801
29.505
30.891
1.00
49.38
W


ATOM
2422
OH2
TIP
A
54
4.648
43.961
2.873
1.00
57.90
W


ATOM
2423
OH2
TIP
A
55
18.093
43.972
14.582
1.00
66.29
W


ATOM
2424
OH2
TIP
A
56
32.762
25.967
29.911
1.00
57.94
W


ATOM
2425
OH2
TIP
A
57
6.942
34.079
7.083
1.00
48.37
W


ATOM
2426
OH2
TIP
A
58
43.391
12.772
6.133
1.00
61.46
W


ATOM
2427
OH2
TIP
A
59
8.265
49.640
−10.664
1.00
59.37
W


ATOM
2428
OH2
TIP
A
60
6.971
18.923
10.395
1.00
62.11
W


ATOM
2429
OH2
TIP
A
61
12.616
28.504
1.451
1.00
57.01
W


ATOM
2430
OH2
TIP
A
62
25.673
48.908
−3.608
1.00
48.26
W


ATOM
2431
OH2
TIP
A
63
11.864
30.819
−2.529
1.00
57.25
W


ATOM
2432
OH2
TIP
A
64
23.756
43.583
−16.811
1.00
53.58
W


ATOM
2433
OH2
TIP
A
65
14.207
19.466
0.982
1.00
52.77
W


ATOM
2434
OH2
TIP
A
66
2.064
35.531
−8.984
1.00
67.76
W


ATOM
2435
OH2
TIP
A
67
27.154
45.880
16.535
1.00
61.40
W


ATOM
2436
OH2
TIP
A
68
29.710
31.367
12.036
1.00
54.10
W


ATOM
2437
OH2
TIP
A
69
37.562
26.637
4.885
1.00
84.16
W


ATOM
2438
OH2
TIP
A
70
16.121
18.617
−9.991
1.00
46.09
W


ATOM
2439
OH2
TIP
A
1
28.748
17.651
−3.973
1.00
43.52
SS


ATOM
2440
OH2
TIP
A
2
40.158
21.998
7.668
1.00
43.52
SS


ATOM
2441
OH2
TIP
A
3
20.062
48.816
5.351
1.00
43.52
SS


ATOM
2442
OH2
TIP
A
4
24.944
5.885
−0.742
1.00
43.52
SS


ATOM
2443
OH2
TIP
A
5
30.688
25.005
−4.289
1.00
43.52
SS


ATOM
2444
OH2
TIP
A
6
19.073
30.215
−9.776
1.00
43.52
SS


ATOM
2445
OH2
TIP
A
7
18.367
30.661
2.655
1.00
43.52
SS


ATOM
2446
OH2
TIP
A
8
19.367
32.272
0.565
1.00
43.52
SS


ATOM
2447
OH2
TIP
A
9
29.275
23.177
−0.801
1.00
43.52
SS


ATOM
2448
OH2
TIP
A
10
30.926
16.038
−5.142
1.00
43.52
SS


ATOM
2449
OH2
TIP
A
11
34.325
28.514
11.449
1.00
43.52
SS


ATOM
2450
OH2
TIP
A
12
21.383
27.492
−13.294
1.00
43.52
SS


ATOM
2451
OH2
TIP
A
13
22.108
28.693
11.655
1.00
43.52
SS


ATOM
2452
OH2
TIP
A
14
18.252
23.094
−9.262
1.00
43.52
SS


ATOM
2453
OH2
TIP
A
15
18.047
20.583
−11.156
1.00
43.52
SS


ATOM
2454
OH2
TIP
A
16
17.057
52.115
−4.118
1.00
43.52
SS


ATOM
2455
OH2
TIP
A
17
22.954
25.044
−0.471
1.00
43.52
SS


ATOM
2456
OH2
TIP
A
18
30.914
51.700
1.620
1.00
43.52
SS


ATOM
2457
OH2
TIP
A
19
18.868
45.761
14.958
1.00
43.52
SS


ATOM
2458
OH2
TIP
A
20
11.192
43.417
6.627
1.00
43.52
SS


ATOM
2459
OH2
TIP
A
21
21.395
33.255
9.085
1.00
43.52
SS


ATOM
2460
OH2
TIP
A
22
31.566
40.059
−1.449
1.00
43.52
SS


ATOM
2461
OH2
TIP
A
23
26.608
40.843
12.469
1.00
43.52
SS


ATOM
2462
OH2
TIP
A
24
10.882
49.731
3.097
1.00
43.52
SS


ATOM
2463
OH2
TIP
A
25
24.112
6.394
−4.916
1.00
43.52
SS


ATOM
2464
OH2
TIP
A
26
20.065
29.530
9.976
1.00
43.52
SS


ATOM
2465
OH2
TIP
A
27
26.915
45.385
9.401
1.00
43.52
SS


ATOM
2466
OH2
TIP
A
28
16.232
36.475
12.662
1.00
43.52
SS


ATOM
2467
OH2
TIP
A
29
17.678
25.766
6.317
1.00
43.52
SS


ATOM
2468
OH2
TIP
A
30
20.570
25.957
0.315
1.00
43.52
SS


ATOM
2469
OH2
TIP
A
31
8.561
24.058
9.246
1.00
43.52
SS


ATOM
2470
OH2
TIP
A
32
20.329
24.123
6.075
1.00
43.52
SS


ATOM
2471
OH2
TIP
A
33
25.927
42.053
15.359
1.00
43.52
SS


ATOM
2472
OH2
TIP
A
34
33.530
38.867
−7.089
1.00
43.52
SS


ATOM
2473
OH2
TIP
A
35
35.591
12.620
10.017
1.00
43.52
SS


ATOM
2474
OH2
TIP
A
36
31.225
41.351
6.010
1.00
43.52
SS


ATOM
2475
OH2
TIP
A
37
22.123
24.472
11.325
1.00
43.52
SS


ATOM
2476
OH2
TIP
A
38
28.087
29.403
−13.302
1.00
43.52
SS


ATOM
2477
OH2
TIP
A
39
34.560
12.999
5.277
1.00
43.52
SS


ATOM
2478
OH2
TIP
A
40
9.465
48.542
10.702
1.00
43.52
SS


ATOM
2479
OH2
TIP
A
41
12.503
38.149
20.364
1.00
43.52
SS


ATOM
2480
OH2
TIP
A
42
7.420
41.639
10.909
1.00
43.52
SS


ATOM
2481
OH2
TIP
A
43
35.680
12.866
7.902
1.00
43.52
SS


ATOM
2482
OH2
TIP
A
44
39.502
27.931
18.775
1.00
43.52
SS


ATOM
2483
OH2
TIP
A
45
21.146
25.021
8.714
1.00
43.52
SS


ATOM
2484
OH2
TIP
A
46
30.337
39.275
−10.268
1.00
43.52
SS


ATOM
2485
OH2
TIP
A
47
18.733
12.572
4.205
1.00
43.52
SS


ATOM
2486
OH2
TIP
A
48
8.456
43.720
10.493
1.00
43.52
SS


ATOM
2487
OH2
TIP
A
49
21.738
21.295
7.389
1.00
43.52
SS


ATOM
2488
OH2
TIP
A
50
20.445
28.671
−16.154
1.00
43.52
SS


ATOM
2489
OH2
TIP
A
51
22.600
12.989
−4.498
1.00
43.52
SS


ATOM
2490
OH2
TIP
A
52
36.376
34.961
13.547
1.00
43.52
SS


ATOM
2491
OH2
TIP
A
53
42.964
28.274
−7.367
1.00
43.52
SS


ATOM
2492
OH2
TIP
A
54
17.026
16.386
5.402
1.00
43.52
SS


ATOM
2493
OH2
TIP
A
55
20.135
54.487
2.295
1.00
43.52
SS


ATOM
2494
OH2
TIP
A
56
30.869
37.274
6.313
1.00
43.52
SS


ATOM
2495
OH2
TIP
A
57
33.853
40.105
6.671
1.00
43.52
SS


ATOM
2496
OH2
TIP
A
58
16.349
43.334
14.737
1.00
43.52
SS


ATOM
2497
OH2
TIP
A
59
18.622
23.529
3.183
1.00
43.52
SS


ATOM
2498
OH2
TIP
A
60
26.630
48.674
7.609
1.00
43.52
SS


ATOM
2499
OH2
TIP
A
61
16.555
47.864
−8.705
1.00
43.52
SS


ATOM
2500
OH2
TIP
A
62
23.193
10.211
−8.266
1.00
43.52
SS


ATOM
2501
OH2
TIP
A
63
6.761
33.422
13.017
1.00
43.52
SS


ATOM
2502
OH2
TIP
A
64
31.235
43.998
6.574
1.00
43.52
SS


ATOM
2503
OH2
TIP
A
65
12.837
27.261
−2.851
1.00
43.52
SS


ATOM
2504
OH2
TIP
A
66
7.112
44.502
1.340
1.00
43.52
SS


ATOM
2505
OH2
TIP
A
67
41.572
21.679
−0.685
1.00
43.52
SS


ATOM
2506
OH2
TIP
A
68
22.680
39.284
19.173
1.00
43.52
SS


ATOM
2507
OH2
TIP
A
69
39.463
37.601
−3.326
1.00
43.52
SS


ATOM
2508
OH2
TIP
A
70
27.198
44.213
12.911
1.00
43.52
SS


ATOM
2509
OH2
TIP
A
71
20.278
32.081
11.495
1.00
43.52
SS


ATOM
2510
OH2
TIP
A
72
18.798
23.783
14.243
1.00
43.52
SS


ATOM
2511
OH2
TIP
A
73
29.802
13.025
4.755
1.00
43.52
SS


ATOM
2512
OH2
TIP
A
74
10.477
43.339
−5.465
1.00
43.52
SS


ATOM
2513
OH2
TIP
A
75
36.280
13.938
−3.506
1.00
43.52
SS


ATOM
2514
OH2
TIP
A
76
17.052
11.701
−7.565
1.00
43.52
SS


ATOM
2515
OH2
TIP
A
77
18.133
24.995
−0.393
1.00
43.52
SS


ATOM
2516
OH2
TIP
A
78
38.072
39.296
5.240
1.00
43.52
SS


ATOM
2517
OH2
TIP
A
79
25.945
15.534
−14.313
1.00
43.52
SS


ATOM
2518
OH2
TIP
A
80
15.465
37.437
19.549
1.00
43.52
SS


ATOM
2519
OH2
TIP
A
81
16.620
20.634
1.922
1.00
43.52
SS


ATOM
2520
OH2
TIP
A
82
23.545
39.853
−13.805
1.00
43.52
SS


ATOM
2521
OH2
TIP
A
83
20.253
35.091
−10.327
1.00
43.52
SS


ATOM
2522
OH2
TIP
A
84
36.698
24.059
13.386
1.00
43.52
SS


ATOM
2523
OH2
TIP
A
85
13.944
26.765
1.654
1.00
43.52
SS


ATOM
2524
OH2
TIP
A
86
3.518
41.699
1.425
1.00
43.52
SS


ATOM
2525
OH2
TIP
A
87
23.552
46.329
−1.900
1.00
43.52
SS


ATOM
2526
OH2
TIP
A
88
21.467
19.279
−8.361
1.00
43.52
SS


ATOM
2527
OH2
TIP
A
89
11.984
27.072
27.543
1.00
43.52
SS


ATOM
2528
OH2
TIP
A
90
23.521
11.079
23.940
1.00
43.52
SS


ATOM
2529
OH2
TIP
A
91
28.454
48.510
−3.162
1.00
43.52
SS


ATOM
2530
OH2
TIP
A
92
13.904
43.690
16.624
1.00
43.52
SS


ATOM
2531
OH2
TIP
A
93
7.949
31.840
5.015
1.00
43.52
SS


ATOM
2532
OH2
TIP
A
94
32.500
19.725
14.392
1.00
43.52
SS


ATOM
2533
OH2
TIP
A
95
38.390
28.352
−11.079
1.00
43.52
SS


ATOM
2534
OH2
TIP
A
96
33.535
30.231
18.680
1.00
43.52
SS


ATOM
2535
OH2
TIP
A
97
42.143
19.333
8.275
1.00
43.52
SS


ATOM
2536
OH2
TIP
A
98
14.934
50.010
−7.142
1.00
43.52
SS


ATOM
2537
OH2
TIP
A
99
30.152
44.543
−0.015
1.00
43.52
SS


ATOM
2538
OH2
TIP
A
100
8.844
47.803
−7.596
1.00
43.52
SS


ATOM
2539
OH2
TIP
A
101
27.103
37.051
19.604
1.00
43.52
SS


ATOM
2540
OH2
TIP
A
102
17.787
15.577
−9.801
1.00
43.52
SS


ATOM
2541
OH2
TIP
A
104
16.591
25.543
2.635
1.00
43.52
SS


ATOM
2542
OH2
TIP
A
105
27.295
22.133
−0.598
1.00
43.52
SS


ATOM
2543
OH2
TIP
A
106
8.558
49.822
−12.353
1.00
43.52
SS


ATOM
2544
OH2
TIP
A
107
6.922
43.138
−1.316
1.00
43.52
SS


ATOM
2545
OH2
TIP
A
108
42.011
13.562
11.695
1.00
43.52
SS


ATOM
2546
OH2
TIP
A
109
20.749
53.219
5.773
1.00
43.52
SS


ATOM
2547
OH2
TIP
A
110
32.142
24.552
22.465
1.00
43.52
SS


ATOM
2548
OH2
TIP
A
111
13.126
28.687
14.559
1.00
43.52
SS


ATOM
2549
OH2
TIP
A
112
36.222
10.792
2.017
1.00
43.52
SS


ATOM
2550
OH2
TIP
A
113
20.573
36.856
1.414
1.00
43.52
SS


ATOM
2551
OH2
TIP
A
115
27.421
10.974
25.524
1.00
43.52
SS


ATOM
2552
OH2
TIP
A
116
42.751
19.151
10.460
1.00
43.52
SS


ATOM
2553
OH2
TIP
A
117
35.543
36.745
5.730
1.00
43.52
SS


ATOM
2554
OH2
TIP
A
118
11.440
28.831
21.618
1.00
43.52
SS


ATOM
2555
OH2
TIP
A
119
7.319
34.101
9.399
1.00
43.52
SS


ATOM
2556
OH2
TIP
A
121
36.857
26.359
21.626
1.00
43.52
SS


ATOM
2557
OH2
TIP
A
123
36.523
30.443
7.429
1.00
43.52
SS


ATOM
2558
OH2
TIP
A
124
26.132
50.469
5.525
1.00
43.52
SS


ATOM
2559
OH2
TIP
A
126
34.329
44.402
−4.548
1.00
43.52
SS


ATOM
2560
OH2
TIP
A
127
26.999
33.484
−9.280
1.00
43.52
SS


ATOM
2561
OH2
TIP
A
128
18.443
27.124
21.387
1.00
43.52
SS


ATOM
2562
OH2
TIP
A
129
17.596
28.119
10.054
1.00
43.52
SS


ATOM
2563
OH2
TIP
A
131
14.334
35.901
18.626
1.00
43.52
SS


ATOM
2564
OH2
TIP
A
132
31.030
32.454
7.688
1.00
43.52
SS


ATOM
2565
OH2
TIP
A
133
2.973
33.533
13.781
1.00
43.52
SS


ATOM
2566
OH2
TIP
A
137
25.307
9.302
−4.123
1.00
43.52
SS


ATOM
2567
OH2
TIP
A
138
22.308
8.017
22.539
1.00
43.52
SS


ATOM
2568
OH2
TIP
A
140
21.574
38.666
3.205
1.00
43.52
SS


ATOM
2569
OH2
TIP
A
146
31.586
31.424
10.186
1.00
43.52
SS


ATOM
2570
OH2
TIP
A
154
6.490
36.861
3.474
1.00
43.52
SS


ATOM
2571
OH2
TIP
A
156
12.899
47.499
−10.970
1.00
43.52
SS


ATOM
2572
OH2
TIP
A
160
38.133
32.578
21.040
1.00
43.52
SS


ATOM
2573
OH2
TIP
A
185
16.312
28.123
22.475
1.00
43.52
SS


ATOM
2574
OH2
TIP
A
191
34.865
30.182
−7.396
1.00
43.52
SS


ATOM
2575
OH2
TIP
A
198
38.682
29.942
21.401
1.00
43.52
SS


ATOM
2576
OH2
TIP
A
221
20.664
28.062
−4.027
1.00
43.52
SS


ATOM
2577
OH2
TIP
A
227
38.426
34.659
9.615
1.00
43.52
SS


TER


END

## Claims

1. An isolated and purified non-crystalline aromatic prenyltransferase having a beta/alpha barrel structure and at least 95% sequence identity with the amino acid sequence set forth in SEQ ID NO:2.

2. An aromatic prenyltransferase according to claim 1, wherein said aromatic prenyltransferase has the amino acid sequence set forth in SEQ ID NO:2.

3. A method of identifying a potential inhibitor of the activity of an aromatic prenyltransferase according to claim 1, said method comprising:
contacting said aromatic prenyltransferase with a potential compound; and
determining the ability of said compound to inhibit the activity of said aromatic prenyltransferase, whereby those compounds which inhibit the activity of said aromatic prenyltransferase are identified as inhibitors of the activity of said aromatic prenyltransferase.

4. A composition comprising an aromatic prenyltransferase consisting of the amino acid seguence of SEQ ID NO:2 in crystalline form, wherein the crystalline form of said aromatic prenyltransferase has space group P21212, with average unit cell dimensions of a=71 Å, b=92 Å, c=48 Å, α=β=γ=90°.

5. A composition according to claim 4, wherein the crystalline form of said aromatic prenyltransferase contains one monomer per asymmetric unit and a solvent content of 45%.

6. A composition according to claim 4, wherein said prenyltransferase having the structural coordinates set forth in Appendix 1.

7. A composition according to claim 5 having the structural coordinates set forth in Appendix 1.