PATENT DOCUMENT

Abstract:
Soluble polypeptide fraction consisting of all or part of one at least one of the four immunoglobulin-type extracellular LAG-3 protein domains (amino acids 1-159, 160-230, 240-330 and 331-412 of the SEQ ID NO:1 sequence) or consisting of one peptide sequence derived from these domains by replacement, addition or deletion of one or more amino acids. The fraction of the invention has a specificity at least equal to that of LAG-3 in relation to its ligand.

Full Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application is a division of application Ser. No. 08/737,271, filed Dec. 24, 1996, now U.S. Pat. No. 5,955,300, which is a §371 application of PCT/FR95/00593, filed May 5, 1995. 
    
    
     BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The invention relates to soluble forms derived from the LAG-3 membrane protein which are useful as immunosuppressants, as well as antibodies capable of preventing the specific binding of the LAG-3 protein to MHC (major histocompatibility complex) Class II molecules as immunostimulants. 
     2. Description of the Related Art 
     In WO-A 91/10682, a protein designated LAG-3 has been described. 
     The LAG-3 protein is a protein selectively expressed by NK cells and activated T lymphocytes. Similarity of the amino acid sequence, the comparative exon/intron organization and the chromosomal localization shown that LAG-3 is related to CD4. The initial characterization of the LAG-3 gene has been described by TRIEBEL et al. (1). 
     The corresponding DNA codes for a type I transmembrane protein of 498 amino acids containing 4 extracellular sequences of the immunoglobulin type. LAG-3 is a member of the immunoglobulin superfamily. 
     The mature protein comprises 476 amino acids (SEQ ID NO:1) with a theoretical molecular weight of 52 kD. The extracellular region contains 8 cysteine residues and 4 potential N-glycosylation sites. By Western blot analysis, it was shown that LAG-3 inside PHA-blasts or activated NK cells has an apparent mass Mr of 70,000. After treatment with N-glycosidase F, a reduction in size to 60 kD was obtained, thereby demonstrating that native LAG-3 is glycosylated. Fuller details are described in WO-A 91/10682. 
     BAIXERAS et al., in J. Exp. Med. 176, 327-337 (2), have, in addition, described their finding that rosette formation between cells transfected with LAG-3 (expressing LAG-3 at their surface) and B lymphocytes expressing MHC Class II was specifically dependent on LAG-3/MHC Class II interaction. 
     Surprisingly, this ligand for MHC Class II was detected with higher levels on activated CD8 +   lymphocytes (MHC Class I-restricted) than on activated CD4 +   lymphocytes. In vivo, only a few disseminated LAG-3 +   cells (MHC Class II-restricted) were to be found in non-hyperplastic lymphoid tissue comprising the primary lymphoid organs, that is to say thymus and bone marrow. LAG-3 +   cells were to be found in hyperplastic lymphoid nodules and tonsils, as well as among peripheral blood mononuclear cells (PBMC) of patients receiving injections of high doses of IL-2. 
     These observations confirm that LAG-3 is an activation antigen in contrast to CD4 expressed in an subpopulation of resting lymphocytes and other cell types, in particular macrophages. 
     The MHC comprises Class I and Class II molecules which are membrane glycoproteins which present fragments of protein antigens to the T lymphocyte receptors (TCR). Class I molecules are responsible for the presentation to CD8 +   cytotoxic cells of peptides derived in large part from endogenously synthesized proteins, while Class II molecules present to CD4 +   helper lymphocytes peptides originating in the first place from foreign proteins which have entered the endocytic, that is to say exogenous, pathway. T helper lymphocytes regulate and amplify the immune response, while cytotoxic lymphocytes are needed to destroy cells irrespective of the tissues expressing &#34;non-self&#34; antigens, for example viral antigens. The mechanism of recognition involves intracellular signals leading to an effective activity of T lymphocytes. 
     It is apparent that, to initiate an immune response mediated by T (CD4 + ) lymphocytes, the foreign antigens must be captured and internalized in the form of peptides by specialized cells, the antigen presenting cells (APC). The resulting antigenic peptides are re-expressed at the surface of the antigen presenting cells, where they are combined with MHC Class II molecules. This MHC Class II/peptide complex is specifically recognized by the T lymphocyte receptor, resulting in an activation of the T helper lymphocytes. 
     Moreover, animal models created by recombination techniques have made it possible to emphasize the part played in vivo by MHC Class II molecules and their ligands. 
     Thus, mice deficient in MHC Class II molecules (3) and possessing almost no peripheral CD4 +   T lymphocytes and having only a few immature CD4 +   lymphocytes in the thymus have proved to be completely incapable of responding to T-dependent antigens. 
     CD4 -  / -   mutant mice (4) have a substantially decreased T lymphocyte activity but show normal development and function of the CD8 +   T lymphocytes, demonstrating that the expression of CD4 on the daughter cells and CD4 +   CD8 +   thymocytes is not obligatory for the development. Compared to normal mice, these CD4-deficient mice have a large amount of CD4 -   CD8 -   cells. 
     These doubly negative cells are restricted to MHC Class II and capable of recognizing the antigen. 
     When they are infected with Leishmania, these mice show a population of functional T helper lymphocytes despite the absence of CD4. These cells are restrictive to MHC Class II and produce interferon-γ when they are activated by the antigen. This indicates that the lineage of the T lymphocytes and their peripheral function need not necessarily depend on the function of CD4. 
     It is not recognized that the proteins encoded by MHC Class II region are involved in many aspects of immune recognition, including the interaction between different lymphoid cells such as lymphocytes and antigen presenting cells. Different observations have also shown that other mechanisms which do not take place via CD4 participate in the effector function of T helper lymphocytes. 
     These different observations underline the pivotal role played by MHC Class II and its ligands in the immune system. 
     Moreover, the importance is known of chimeric molecules composed of the extracytoplasmic domain of proteins capable of binding to ligands and a constant region of human immunoglobulin (Ig) chains for obtaining soluble forms of proteins and of cell receptors which are useful, in particular, as therapeutic agents. 
     Thus, soluble forms of CD4 have proven their efficacy in inhibiting an HIV infection in vitro in a dose-dependent manner. 
     Nevertheless, clinical trials with soluble CD4 molecules, in particular of CD4-Ig, have not enabled a significant decrease in viral titres to be demonstrated. Transgenic mice expressing up to 20 μg/ml of soluble CD4 in their serum were created. These mice showed no difference as regards their immune function relative to control mice. Hitherto, no direct binding to MHC Class II of molecules derived from CD4 has been reported. This strongly suggests that soluble CD4 molecules do not interact in vivo with MHC Class II molecules. 
     SUMMARY OF THE INVENTION 
     Surprisingly, the authors of the present invention have shown that soluble molecules containing different fragments of the extracytoplasmic domain of the LAG-3 protein were capable of binding to MHC Class II molecules and of having an immunosuppressant action. 
     The extracytoplasmic region of LAG-3 represented by the sequence SEQ ID No. 1 comprises the domains D1, D2, D3 and D4 extending from amino acids 1 to 149, 150 to 239, 240 to 330 and 331 to 412, respectively. 
     Thus, the subject of the invention is a soluble polypeptide fraction consisting of all or part of at least one of the 4 immunoglobulin type extracellular domains of the LAG-3 protein (amino acid 1 to 149, 150 to 239, 240 to 330 and 331 to 412 of the sequence SEQ ID No. 1), or of a peptide sequence derived from these domains by replacement, addition and/or deletion of one or more amino acids, and which possesses a specificity at least equal to or greater than that of LAG-3 for its ligand. 
     The present invention encompasses, in particular, soluble polypeptide fractions having a sequence derived from the native LAG-3 sequence originating from the well-known phenomenon of polytypy. 
     The soluble polypeptide fraction is characterized in that it comprises the peptide region of LAG-3 responsible for the affinity of LAG-3 for MHC Class II molecules. 
     The soluble polypeptide fraction comprises, in particular, a peptide sequence derived from these domains by replacement, addition and/or deletion of one or more amino acids, and which possesses a specificity equal to or greater than that of LAG-3 for its ligand, for example the whole of the first two immunoglobulin type domains of LAG-3, or the 4 immunoglobulin type domains of the extracytoplasmic domain of LAG-3. 
     Advantageously, the soluble polypeptide fraction is comprises of all or part of at least one of the four immunoglobulin type extracellular domains of the LAG-3 protein (amino acid 1 to 149, 150 to 239, 240 to 330 and 331 to 412 of sequence SEQ ID N o  1) comprising one or more of the arginine (Arg) rests at the positions 73, 75 and 76 of sequence SEQ ID N o  1 substituted with glutamic acid (Glu). 
     Preferably, the soluble polypeptide fraction comprises a loop in which the average position of the atoms forming the basic linkage arrangement is given by the position of amino acids 46 to 77 (SEQ ID No. 1) appearing in Table 1 or Table 2 or differs therefrom by not more than 5%. 
     The soluble polypeptide fraction advantageously comprises, in addition, the second immunoglobulin type extracellular domain (D2) of LAG-3 (amino acids 150 to 241). 
     Advantageously, the soluble polypeptide fraction comprises, besides the peptide sequence of LAG-3 as defined above, a supplementary peptide sequence at its C-terminal and/or N-terminal end, so as to constitute a fusion protein. The term &#34;fusion protein&#34; means a portion of any protein permitting modification of the physico-chemical features of the subfragments of the extracytoplasmic domain of the LAG-3 protein. Examples of such fusion properties contain fragments of the extracytoplasmic domain of LAG-3 as are defined above, bound to the heavy chain --CH2--CH3 junction region of a human immunoglobulin, preferably an isotype IgG4 immunoglobulin. 
     Such fusion proteins may be dimeric or monomeric. These fusion proteins may be obtained by recombination techniques well known to a person skilled in the art, for example a technique such as that described by Traunecker et al. (5). 
     Generally speaking, the method of production of these fusion proteins comprising an immunoglobulin region fused with a peptide sequence of LAG-3 as defined above consists in inserting into a vector the fragments of cDNA coding for the polypeptide regions corresponding to LAG-3 or derived from LAG-3, where appropriate after amplification by PCR, and the cDNA coding for the relevant region of the immunoglobulin, this cDNA being fused with cDNA coding for the corresponding polypeptide regions or derivatives of LAG-3, and in expressing after transfection the fragments cDNA in an expression system, in particular mammalian cells, for example hamster ovary cells. 
     The fusion proteins according to the invention may also be obtained by cleavage of a LAG-3/Ig conjugate constructed so as to contain a suitable cleavage site. 
     The subject of the invention is also a therapeutic composition having immunosuppressant activity comprising a soluble polypeptide fraction according to the invention. This composition will be useful for treating pathologies requiring immunosuppression, for example autoimmune diseases. 
     The subject of the invention is also the use of antibodies directed against LAG-3 or soluble polypeptide fractions derived from LAG-3 as are defined above, or fragments of such antibodies, in particular the Fab, Fab&#39; and F(ab&#39;) 2  fragments, for the preparation of a therapeutic composition having immunostimulatory activity. &#34;Immunostimulatory&#34; means a molecular entity capable of stimulating the maturation, differentiation, proliferation and/or function of cells expressing LAG-3, that is to say T lymphocytes or active NK cells. The anti-LAG-3 antibodies may be used as potentiators of vaccines or immunostimulants in immunosuppressed patients, such as patients infected with HIV or treated with immunosuppressant substances, or be used to stimulate the immune system by elimination of self cells displaying abnormal behavior, for example cancer cells. 
     Immunostimulatory activity of anti-LAG-3 antibodies is surprising, inasmuch as anti-CD4 antibodies have an immunosuppressant action. 
     Such antibodies may be polyclonal or monoclonal; however, monoclonal antibodies are preferred. The polyclonal antibodies may be prepared according to well-known methods, such as that described by BENEDICT A. A. et al. (6). Monoclonal antibodies are preferred, on account of the fact that they are specific for a single epitope and yield results with better reproducibility. Methods of production of monoclonal antibodies are well known from the prior art, especially the one described by KOHLER and MILSTEIN. This method, together with variants thereof, are described by YELTON et al. (7). 
     The subject of the invention is also anti-idio-type antibodies directed against the antibodies according to the invention, which contain the internal image of LAG-3 and are consequently capable of binding to MHC Class II. Such antibodies may be used, in particular, as immunosuppressants, and, for example, in autoimmune pathologies. 
     The therapeutic compositions according to the present invention comprise soluble LAG-3 proteins or antibodies as are defined above, as well as a pharmaceutically acceptable vehicle. These compositions may be formulated according to the usual techniques. The vehicle can vary in form in accordance with the chosen administration route; oral, parenteral, sublingual, rectal or nasal. 
     For the compositions of parenteral administration, the vehicle will generally comprise sterile water as well as other possible ingredients promoting the solubility of the composition or its ability to be stored. The parenteral administration routes can consist of intravenous, intramuscular or subcutaneous injections. 
     The therapeutic composition can be of the sustained-release type, in particular for long-term treatments, for example in autoimmune diseases. The dose to be administered depends on the subject to be treated, in particular on the capacity of his/her immune system to achieve the desired degree of protection. The precise amounts of active ingredient to be administered may be readily determined by the practitioner who will initiate the treatment. 
     The therapeutic compositions according to the invention can comprise, in addition to soluble LAG-3 or the antibodies according to the invention, another active ingredient, where appropriate bound via a chemical bond to LAG-3 or to an antibody according to the invention. As an example, there may be mentioned soluble LAG-3 proteins according to the invention fused to a toxin, for example ricin or diphtheria toxoid, capable of binding to MHC Class II molecules and of killing the target cells, for example leukaemic or melanoma cells, or fused to a radioisotope. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIG. 1 shows a comparison of the proliferation of cells incubated with F(ab) fragments of 17B4 to the proliferation of T cells incubated with intact 17B4 monoclonal antibody. 
     FIG. 2 shows the proliferation of Clone 28 in response to tetanus toxoid when co-cultured with 17B4 or control antibody 10H3. 
     FIG. 3 shows the expression vector pCDM7 used for manufacturing the recombinant LAG-3 proteins and a recombinant CD8 immunoadhesin control. 
     FIG. 4 shows the pCLH3 AXS DHFR hα IVS vector used to express amplified LAG-3 sequences. 
     FIG. 5A shows the inhibition of MHC Class II interaction with LAG-3 by recombinant LAG-3 D1-D4 and FIG. 5B shows the potential inhibition of MHC Class II interaction with CD4. 
     FIG. 6 shows the inhibition of Clone 28 proliferation by recombinant LAG-3 D1-D4. 
     FIG. 7 shows the inhibition of Clone 154 proliferation by LAG-3 Ig. 
     FIG. 8 shows the binding of LAG-3 to B cell lines expressing MHC class II haplotypes or human class II-transfected mouse cells. 
     FIG. 9 shows the binding of LAG-3 Ig to MHC Class II expressing Daudi cells. 
     FIG. 10 shows that preincubation of HLA class II expressing cells with 17B4 inhibiting LAG-3 Ig binding. 
     FIG. 11A-11C show the inhibition of clone T154 proliferation by crosslinked LAG-3 Ig. 
     FIG. 12 compares inhibition of T cell proliferation by anti-Class II antibodies to inhibition of T cell proliferation by LAG-3 Ig. 
     FIG. 13A-13D show that T cell proliferation in response to OKT3 (FIG. 13A), lectins (FIG. 13B), and low-concentration IL 2  (FIG. 13C) is inhibited by LAG-3 Ig but proliferation in response to high-concentration IL 2  is not inhibited by LAG-3 Ig (FIG. 13D). 
     FIG. 14 shows clone S1B5 cytotoxicity towards Epstein-Barr virus transformed human B cells. 
     FIG. 15 shows peripheral blood lymphocyte cytotoxicity towards HLA Class I -   Daudi cells. 
    
    
     DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     The examples which follow, together with the attached reference figures, will illustrate the invention in greater detail. 
     EXAMPLE 1 
     Proliferation of active T lymphocyte lines in the presence of anti-LAG-3 monoclonal antibodies 
     The anti-LAG-3 monoclonal antibodies used were 17B4, described in BAIXERAS et al. (2) and deposited at the CNCM under NO. I-1240 on Jul. 10, 1992, and 11E3, described in HUARD et al. (8). 
     These antibodies belong to the isotype IgG1. These antibodies were tested for their biological effects on activated T lymphocytes, stimulated by specific antigenic peptides or processed antigens presented by MHC Class II molecules expressed by autologous antigen presenting cells, expressing LAG-3. 
     An anti-CD48 monoclonal antibody designated 10 H3 was used as irrelevant IgG1 antibody (negative control). 
     The saturating concentrations of anti-LAG-3 and anti-CD48 antibodies were determined by immunofluorescence on PHA (phytohaemagglutinin)-blasts and cell lines transformed by Epstein-Barr virus (EBV). In the proliferation tests, the monoclonal antibodies were added in the proportion of 5 times the saturating concentration. 
     The T lymphocyte lines used were, on the one hand the clone 154 derived from peripheral blood lymphocytes, raised against a peptide mimicking an influenza haemagglutinin (HA) fragment having an amino acid sequence extending from amino acid 306 to 329 (p20 peptide), and on the other hand the clone 28, a T lymphocyte clone derived from peripheral lymphocytes of a single human donor, raised against diphtheria toxoid (DT). The antigen presenting cells (APC) corresponding to clone 154 were EBV-transformed B lymphocytes of the same donor (DR3/DR11) as T 154. The antigen presenting cells corresponding to clone 28 were EBV-transformed B lymphocytes of the same donor. This clone was restricted to HLA DR7. 
     For clone 154, the APC (5×10 6 ) were incubated at 37° C. for one and a half hours with variable doses of the p20 peptide, then washed and irradiated (10,000 rad). The cells were plated out on 96-well microtitration plates at the same time as the clone 154 cells (0.5×10 5  to 10×10 5  cells/ml) in a 3:1 ratio. For clone 28, the responding cells/stimulating cells ratio was 1. 
     The HLA DR7/EBV APC cells were either treated with mitomycin or irradiated, then added to the T lymphocytes in the presence of DT (which remained in the culture). The final concentration of clone 28 cells was 100,000 cells/ml. 
     [ 3  H]Thymidine (1 μCi/well) was added at varying time intervals from day 2 to day 10 of culture. 
     Each experiment was carried out in triplicate. 
     The results were expressed as the mean cpm and after subtraction of the cpm found in the negative control (T lymphocytes cocultured with APC unladen with immunogens). The proliferation tests were carried out on 96-well plates. The absorption of tritiated thymidine in the individual 200 μl wells were measured after adding 1 μCi of thymidine for the last 18 hours of culture. The results were expressed in the form of the mean of 3 tests. The standard deviation was usually less than 12% (a little more in the case of ovary low cpm measurements). Moreover, mixed culture (clone 154/APC) supernatants were combined, filtered through 0.22 μm membranes, divided into samples and frozen at -20° C. until the time of titration using commercial immunoassay kits: Immunotech IL-2 and INF-α titration kit, Genzyme IFN-γ kit and Cayman Chemicals IL-4 kit. 
     A dose determination study was carried out to establish the proliferation profiles of clone 154 brought into contact with the p20 specific antigen at varying concentrations and in the presence or absence of anti-LAG-3 monoclonal antibodies or irrelevant monoclonal antibodies (negative control). 
     The individual results of 16 separate tests showed that, irrespective of the concentration of added antigen, the initial point up to the peak of proliferation was not modified, but a significant prolongation of the proliferation of T lymphocytes incubated with the anti-LAG-3 monoclonal antibodies was observed systematically. Fab fragments of the monoclonal antibody 17B4 were prepared and used in a test of proliferation of clone 154. The proliferation profile of T lymphocytes activated by the antigen with the 17B4 Fab fragments (15 μg/ml) was similar to that of cells incubated in the presence of whole 17B4 monoclonal antibody (40 μg/ml) (FIG. 1). These results show that the observed biological effects are not attributable to a non-specific reaction induced by the Fc region of the anti-LAG-3 monoclonal antibodies. 
     Similar results were obtained with the 11E3 anti-LAG-3 monoclonal antibodies. 
     Clone 28 was also stimulated with the antigen (tetanus toxoid 10 μg/ml) in the presence of 17B4 monoclonal antibodies after coculture with the corresponding APC in the presence of DT. The results are shown in FIG. 2. 
     The effects of the anti-LAG-3 monoclonal antibodies observed with clone 28, namely the prolongation of proliferation, are similar to those observed with clone 154. 
     Tests were carried out designed to measure the miscellaneous cellular events occurring after the antigenic stimulation of clone 154 cells incubated in the presence of anti-LAG-3 monoclonal antibodies. 
     The cells were harvested during conventional antigenic stimulation of clone 154 in the presence of anti-LAG-3 or anti-CD48 monoclonal antibodies or in the absence of antibodies, and tested for the expression of LAG-3 and CD25 transmembrane receptors, and samples of culture supernatants were collected at different time intervals after stimulation and tested for the presence of IFN-γ, TNF-α, IL-4 and IL-2. 
     Two-colour direct immunofluorescence tests (anti-CD3 monoclonal antibodies and anti-CD25 monoclonal antibodies) showed that IL-2 receptors were weakly but significantly increased 5 days after the antigenic stimulation. Similar tests with anti-CD3 and 11E3 (anti-LAG-3) monoclonal antibodies showed that LAG-3 was overexpressed from the day following activation onwards. In addition, the secretion of IL-2, IL-4, IFN-γ and TNF-α was also modulated by incubation with anti-LAG-3 monoclonal antibodies, thus showing that different cellular events are modified by the presence of anti-LAG-3 monoclonal antibodies and that some events already take place 24 hours after stimulation. 
     These results show indirectly that LAG-3 plays a regulatory role for CD 4+   cells. The fact that anti-LAG-3 monoclonal antibodies increase proliferation, and hence act as immunopotentiators, suggest that LAG-3 is involved in the &#34;deactivation&#34; of CD4 +   T lymphocytes with a negative role of LAG-3 on the antigen-dependent stimulation. 
     EXAMPLE 2 
     Transient expression of LAG-3 fusion proteins 
     Soluble proteins derived from LAG-3 were obtained by a recombinant DNA technique using suitable vectors comprising DNA coding for LAG-3 and DNA coding for an immunoglobulin fragment. The transient expression system consisted of transfected Cos cells. This system makes it possible to produce several mg of recombinant fusion proteins. Recombinant DNA techniques were carried out as described by MANIATIS et al. (22). The modifications were made as recommended by the manufacturer. 
     Construction of LAG-3 D1-D4 Ig and LAG-3 D1D2 Ig 
     Fragments coding for the D1D2 or D1-D4 regions were amplified (30 cycles) from a fragment of cDNA (FDC sequence) encompassing LAG-3 cDNA (TRIEBEL et al., (1)), using Taq polymerase free from 5&#39;-endonuclease activity and relatively resistant to an exposure to very high temperature; the amplification was followed by a denaturation at 98° C. (with a Perkin Elmer Cetus &#34;DNA thermal cycle&#34;). Specific primers were used as recorded in the table below. 
     The resulting amplified fragments (739 bp and 1312 bp for LAG-3 D1D2 and LAG-3 D1-D4, respectively) were inserted into a pBS plasmid (Stratagene). 
     Inserts were prepared after digestion with XhoI and BglII and introduced into the XhoI/BamHI sites of the vector pCDM7-CD8-IgG1 (pCDM7 being derived from pCDM8 marketed by Stratagene), as illustrated in FIG. 3, so as to exchange the DNA sequences coding for CD8 for those coding for the subfragments of LAG-3. The resulting expression vectors contained the sequences coding for D1D2 or D1-D4 fused to the DNA sequences coding for the --CH 2  --CH 3  junction region of a human IgG1 chain. 
     
                                           TABLE 3__________________________________________________________________________Primers used to amplify LAG-3 DNA sequences by PCR                                Resulting                                encoded subfrag-                                ment fused withPrimers used for amplification of the DNA                                                    a subfragment__________________________________________________________________________                                IgPrimer (5&#39;)                          LAG-3 D1D25&#39; GCGCCTCGAGGCCCAGACCATAGGAGAGATGT 3&#39; (SEQ ID NO:2)                                          from the leadercoupling   untranslated   start of                              sequence                                tosite       5&#39; sequences   translation                                                        amino acid                                241Primer (3&#39;)5&#39;  GCGCAGATCTCTCCAGACCCAGAACAGTGAGGTTATACAT 3&#39; (SEQ ID                                 O:3)BglII coup-            End of D2ling sitePrimer (5&#39;)                                             LAG-3 D1-D4identical to LAG-3 D1D2Primer (3&#39;)                                             from the leader5&#39; GCGCAGATCTACCTGGGCTAGACAGCTCTGTGAA 3&#39; (SEQ ID NO:4)                                       sequence to                                  amino acid 412BglII coup-            End of D4ling site__________________________________________________________________________ 
    
     CDM7 is a eukaryotic expression vector derived from the vectors developed by SEED et al. (10) for the cloning of DNA and its expression in E. coli and eukaryotic cells. CDM7 possesses the following features: (i) the human cytomegalovirus promoter for transient expression in mammalian cells; (ii) a viral origin of SV40 for an autosomal replication of mammalian cells expressing T antigen; (iii) π VX (type Col E1) as plasmid origin for a high copy number; (iv) a Sup F selection for resistance to ampicillin and tetracycline in Tet amb  and Amp amb  E. coli strains; (v) an origin of replication of M13 for the release of a single strand; (ix) a T7 RNA promoter; and (vii) a polylinker for an efficient cloning of heterologous DNA. 
     Transient expression in Cos cells 
     Cos cells (5×10 6 ) were transfected with 30 μg of DNA of suitable expression vectors (coding for either LAG-3 D1D2 Ig, or LAG-3 D1-D4 Ig, or CD8 Ig) by electroporation (200 V, 1500 μF, 30-40 msec) using a Cellject apparatus (Eurogentech, Liege, BE). The cells were plated out again and cultured on a medium containing 5% of foetal calf serum. The supernatants were withdrawn 6 days after transfection. 
     The synthesis of the resulting fusion proteins was analysed from the supernatants as well as from cell extracts of transfected cells, by Western blot analysis with the 17B4 monoclonal antibodies. Immunoreactive materials were observed in the supernatant of cells transfected with DNA coding for LAG-3 D1D2 Ig or LAG-3 D1-D4 Ig. 
     Concomitantly, a recombinant CD8 immunoadhesin (CD8 Ig) was obtained as negative control using the same expression system and the expression vector pCDM7-CD8 (FIG. 3). 
     The recombinant proteins LAG-3 D1D2 Ig, LAG-3 D1-D4 and CD8 Ig were purified by means of the standard method on protein A-Sepharose. The resulting material was analysed by SDS-PAGE, followed by Coomassie staining or a Western blot analysis using anti-human Ig antibody. 
     EXAMPLE 3 
     Production of soluble subfragments of LAG-3 
     In order to produce large amounts of recombinant proteins, a stable expression system comprising of transfected mammalian cells was developed. The host cells are anchorage-dependent hamster ovary (CHO) cells isolated from CHO cells deficient in dihydrofolate reductase (DHFR) and consequently necessitating glycine, a purine and thymidine for their growth. The pivotal role of DHFR in the synthesis of nucleic acid precursors, combined with the sensitivity of DHFR-deficient cells with respect to tetrahydrofolate analogues such as methotrexate (MTX), has two major advantages. Transfection of these cells with expression vectors containing the DHFR gene permits the secretion of recombinant DHFR-resistant clones, and the culturing of these cells on selective media containing increasing amounts of MTX results in amplification of the DHFR gene and the DNA associated therewith. 
     Construction of LAG-3 D1, LAG-3 D1D2, LAG-3 D1-D4 
     Fragments of DNA coding for the D1, D1D2 or D1-D4 regions were amplified using a PCR method identical to the one described previously, using the primers specified in the table below. 
     
                                           TABLE 4__________________________________________________________________________Primers used for amplifying LAG-3 DNA sequences by PCR                              Resulting                                                  encodedPrimers used for amplification of the DNA                                                subfragment__________________________________________________________________________Primer (5&#39;)                        LAG-3 D15&#39; CGCCGTCGACCGCTGCCCAGACCATAGGAGAGATGTG 3&#39; (SEQ ID NO:5)                                                  from the leaderSalI coup-     untranslated  start of                                                    sequence toling site      5&#39; sequences  translation                                                 amino acid 149Primer (3&#39;)5&#39; GCGCGTCGACTTACATCGAGGCCTGGCCCAGGCGCAG 3&#39; (SEQ ID NO:6)SalI coup-     End of D1ling sitePrimer (5&#39;)                                       LAG-3 D1D2identical to LAG-3 D1Primer (3&#39;)                                       from the leader5&#39; GCGCGTCGACTTAACCCAGAACAGTGAGGTTATAC 3&#39; (SEQ ID NO:7)                                  sequence to                                                  amino acid 239SalI coup-     End of D2ling sitePrimer (5&#39;)                                       LAG-3 D1-D4identical to LAG-3 D1Primer (3&#39;)                                       from the leader5&#39; GCGCGTCGACTTAACCTGGGCTAGACAGCTCTGTG 3&#39; (SEQ ID NO:8)                                  sequence to                                                  amino acid 412SalI coup-     End of D4ling site__________________________________________________________________________ 
    
     The resulting amplified fragments were digested with SalI and inserted into the SalI site of pUC 18 (Stratagene). 
     The amplified sequences were verified, and the inserts subcloned into the expression vector pCLH3 AXS V2 DHFR hα IVS as described by COLE et al. (Biotechnology 11, 1014-1024, 1993) (FIG. 4). 
     This vector is a eukaryotic expression vector which is multifunctional for the expression cDNA and its amplification in eukaryotic cells. It possesses the following features: (i) the murine promoter of the metallothionein-1 gene and a polyadenylation sequence SV 40 (comprising a donor-acceptor splicing site) to bring about transcription of the gene of interest, (ii) a human intervening sequence A containing the donor-acceptor splicing site of the gene for the subunit of α glycoprotein for obtaining high levels of transcription of cDNA, (iii) the pML sequence containing the origin of replication of pBR 322 and a gene for resistance to ampicillin for bacterial amplification, and (iv) a DHFR transcription unit of SV 40 to bring about transcription of the sequences used for selection and amplification of the transfectants. 
     Stable expression in CHO cells 
     The expression vectors coding for LAG-3 D1, LAG-3 D1D2 and LAG-3 D1-D4 were used to transfect CHO DUKX cells, and these cells were cultured on a selective medium. Cells capable of multiplying under these conditions were combined and cultured on a medium containing increasing amounts of MTX. Levels of expression were measured by Western blot analysis using the 17B4 monoclonal antibody. Clones producing high levels of recombinant soluble molecules derived from LAG-3 were propagated in bioreactors, and the material derived from LAG-3 was purified by ion exchange chromatography and immunoaffinity. 
     Western blot analyses revealed, in supernatants of cells transfected with expression vectors coding for LAG-3 D1, LAG-3 D1D2 and LAG-3 D1-D4, bands with apparent Mr values of 15 to 18 kD, 34-36 kD (doublets) and 55 kD (2 possible bands). The respective Mr values of these immunoreactive materials corresponded to the expected Mr values of glycosylated LAG-3 D1 Ig (139 amino acids and a putative N-glycosylating site), glycosylated LAG-3 D1D2 Ig (239 amino acids containing 3 glycosylation sites) and glycosylated LAG-3 D1-D4 (412 amino acids containing 4 glycosylation sites). 
     EXAMPLE 4 
     Specific binding of LAG-3 Ig to cells expressing MHC Class II 
     The reactivity of the monoclonal antibodies and of LAG-3 D1-D4 was studied by indirect immunofluorescence. Target cells (4×10 5 ) were incubated for 30 minutes at 4° C. in the presence of LAG-3 D1-D4 Ig, CD8 Ig, a murine monoclonal antibody, (949) anti-human MHC Class II (DR, DP, DQ) conjugated to FITC (isothiocyanate fluoride) from a Coulter clone, or murine Ig-FITC: an irrelevant immunoglobulin G conjugated to FITC. The cells were washed and incubated at 4° C. for 30 minutes with either a goat anti-human Ig polyclonal F(ab&#39;) 2  conjugated to fluorescein or a goat anti-mouse Ig polyclonal antibody conjugated to fluorescein (Coulter clone). 
     To confirm the LAG-3/MHC Class II binding, LAG-3 D1-D4 Ig was incubated with MHC class II-positive or -negative cells. Four B lymphocyte lines expressing MHC Class II(L31, Phil EBV, Raji, Sanchez and Personnaz) were treated with anti-Class II monoclonal antibody 949, or the supernatants of Cos cells transfected with DNA coding either for LAG-3 D1-D4 or for CD8 Ig. The five cell lines expressing the different haplotypes of MHC Class II molecules were recognized by LAG-3 Ig in the same way as by the anti-Class II monoclonal antibodies (positive control), while the supernatant containing CD8 Ig (negative control) did not bind to these cell lines, as could be expected. Four MHC Class II-negative cell lines (CEM, RJ, HSB2, K562) were treated with the same reagents as above. None reacted, either with the anti-MHC Class II (negative control) or with LAG-3 D1-D4 Ig, showing that the binding of LAG-3 D1-D4 is specific to MHC Class II molecules. 
     Further experiments were carried out using (i) mouse fibroblasts transfected or otherwise with genes coding for human DR7 or human DP4, (ii) mouse cells expressing or otherwise MHC Class II molecules, (iii) activated human CD4 +   or CD8 +   cells, and (iv) T lymphocyte lines expressing the different haplotypes of MHC Class II molecules (FIG. 8). 
     Unlike CD8 Ig, LAG-3 D1-D4 Ig binds to all cells expressing MHC Class II as efficiently as the anti-MHC Class II monoclonal antibody 949. LAG-3 D1-D4 Ig binds to all DR and DP haplotypes tested, to human MHC Class II molecules expressed by transfected mouse cells, to murine MHC Class II molecules and also to MHC Class II molecules expressed by CD4 +   or CD8 +   lymphocytes. 
     These results represent for the first time proof that soluble molecules derived from a ligand for MHC Class II are capable of binding to cells expressing MHC Class II. 
     Similar experiments showed that LAG-3 D1D2 bound to cells expressing MHC Class II in as specific a manner and with the same efficiency as LAG-3 D1-D4. 
     Binding activity of LAG-3Ig and cellular distribution of ligands for LAG-3Ig 
     The capacity of this immunoadhesin to bind to cell ligands is measured using a fluorescein-labelled goat serum directed against human immunoglobulins. 
     In these experiments, the target cells are first incubated with a human monoclonal antibody or an immunoadhesin for 30 min at 4° C. in RPMI 1640 containing 10% of FCS (foetal calf serum). The cells are then incubated with an FITC-labelled goat anti-mouse immunoglobulin serum (Coulter) for the murine monoclonal antibodies or with an FITC-labelled goat anti-human immunoglobulin serum (Tago) for the immunoadhesins. The fluorescence is measured after two washes, analyzing 3,000 cells with an Elite cytometer (Coultronics, Hialeah, Fla.). FIG. 9 shows the degrees of binding of LAG-3Ig, CD8Ig, antibody 949 or antibody OKT3 (anti-CD3, ATCC), represented by the number of cells counted as a function of the logarithm of the measured fluorescence intensity. 
     LAG-3Ig binds to mouse fibroblasts transfected for the gene for the HLA DR 4  molecule, and does not bind to untransfected cells. CD8Ig is capable of binding to HLA DR 4   +   fibroblasts under the same conditions. 
     The cellular distribution of the ligands for LAG-3Ig was evaluated on a cell population sample by immunofluorescence. 
     LAG-3Ig is visualized on all positive Class II cells tested, including B cell lines transformed by Epstein-Barr virus (derived from genetically unrelated donors, including 10 homozygous lines of DR 1  to DR 10  typing), as well as on activated T and NK cells. 
     FIG. 9 shows, by way of example, the binding of LAG-3Ig to Daudi cells which are positive for Class II antigens. 
     The means fluorescence intensity with LAG-3Ig is similar to that observed with antibody 949 which is specific for Class II antigens. The binding of LAG-3Ig to DR 4  (FIG. 9), DR 2 , DR 7  and DPw4 (not shown) expressed at the surface of mouse fibroblasts is, in contrast, weaker than that observed for antibody 949. 
     No binding is detected on cell lines which are negative for Class II antigens of T origin (peripheral blood T cells, CEM, HSB2, REX lines), of B origin (RJ 2.2.5 line) or of non-lymphoid origin (human lines, K562 of erythromyoloid origin and line originating from melanoma cells (not shown). 
     Moreover, LAG-3Ig binds to xenogeneic Class II molecules of the MHC, such as the antigens expressed by mouse lymphoma A 20 and the monkey Classes II expressed by phytohaemagglutinin-stimulated blasts (data not shown). 
     The specificity of binding of LAG-3Ig was also verified using the monoclonal antibodies 17B4, whose capacity to block LAG-3/MHC Class II interactions in cell adhesion tests was demonstrated beforehand (FIG. 10). 
     In these experiments, the LAG-3Ig molecules are preincubated for 30 minutes at 4° C. either with medium alone, or with 17B4 (1 mg/ml), or with OKT3 (1 mg/ml), before being brought into contact with Daudi cells. 
     FIG. 10 shows that a preincubation of LAG-3Ig with 17B4 inhibits the binding to Class II +   cells, whereas no inhibition is detected with the OKT3 control. 
     EXAMPLE 5 
     Inhibition of LAG-3/MHC Class II interaction by soluble fragments of LAG-3 
     The inhibition of LAG-3/MHC Class II interaction by the soluble fragments of LAG-3 may be observed directly in relation to the binding of LAG-3Ig by Class II MHC molecules, by competitive experiments with the soluble fragments. 
     To verify whether the soluble LAG-3D 1  D 2  fragments produced by CHO cells could displace the binding of immunoadhesins derived from LAG-3, the following tests were carried out: 
     Daudi cells are incubated with soluble LAG3-D 1  D 2  fragments so as to permit the binding of these molecules to the MHC Class II antigens expressed at the surface of the Daudi cells. 
     In a second step, the cells are incubated in the presence of LAG-3D 1  D 4  Ig in dimeric form or LAG-3D 1  D 2  Ig in monomeric form. 
     The binding of these immunoadhesins derived from LAG-3 is measured using a goat anti-human Ig F(ab&#39;) 2  conjugated to fluorescein (GAH-FITC). 
     The control groups are represented by Daudi cells incubated with dimeric LAG-3D 1  D 4  Ig or monomeric LAG-3D 1  D 2  Ig without preincubation with the soluble LAG-3D 1  D 2  fragments. 
     The results are recorded in Table 5, which shows that the soluble LAG-3D 1  D 2  fragments are capable of displacing the immunoadhesins derived from LAG-3 in mono- or dimeric form. 
     
                       TABLE 5______________________________________                MeanReactants  Detection fluorescence                          Conclusion______________________________________--         GAH-FITC  0.3       GAH does not                          interfereDimeric    GAH-FITC  20.8      The binding ofLAG-3D1D4Ig                    CHO/LAG-3D1D2CHO/LAG-3D1D2,      GAH-FITC  8.5       inhibits the bindingthen dimeric                   of dimericLAG-3D1D4Ig                    LAG-3D1D4Ig (58%)Monomeric  GAH-FITC  62.5      The binding ofLAG-3D1D2Ig                    CHO/LAG-3D1D2CHO/LAG-3D1D2,      GAH-FITC  10.9      inhibits the bindingthen monomeric                 of monomericLAG-3D1D2Ig                    LAG-3D1D2Ig (27%)______________________________________ 
    
     These data confirm that the soluble LAG-3D1D2 fragments bind to MHC Class II molecules. 
     Inhibition of LAG-3/MHC Class II and CD4/MHC Class II interaction 
     Rosette formation between Cos cells transfected with wild-type LAG-3 and B lymphocytes transformed with EBV expressing MHC Class II molecules was demonstrated by BAIXERAS et al. (2). This interaction is inhibited both by anti-LAG-3 and anti-MHC Class II monoclonal antibodies. 
     The method described in this publication was modified by replacing the visualization and counting of Cos cells binding to B lymphocytes by counting the radioactivity remaining after incubation of  51  Cr-labelled B lymphocytes with Cos cells expressing LAG-3 (binding assay). 
     The possible inhibitory effects of soluble molecules derived from LAG-3 on LAG-3/MHC Class II interaction, and also on CD4/MHC Class II interaction, were studied. 
     Cos cells transfected with a suitable expression vector (coding for wild-type LAG-3 or for CD4). Two days later, the Cos cells were treated with trypsin and plated out again on the basis of 0.05×10 6  cells/well on flat-bottomed 12-well tissue culture plates. 24 hours later,  51  Cr-labelled Daudi cells (5.5×10 6 ) were incubated on this monolayer of Cos cells (final vol.: 1 ml) for 1 hour. The target B cells were then aspirated off and the wells washed 5 to 7 times, gently adding 1 ml of medium dropwise. The edges of the wells were washed by suction using a Pasteur pipette. The remaining cells were lysed with 1 ml of PBS, 1% Triton for 15 minutes at 37° C. The lysates were centrifuged at 3000 rpm for 10 minutes, and 100 μl of the resulting supernatant were counted. 
     LAG-3 D1-D4 Ig was used to inhibit LAG-3/MHC Class II and CD4/MHC Class II interaction in the  51  Cr binding assay. Human CD8 Ig and IgG1 were tested in parallel and used as negative controls. 
     A significant inhibition of LAG-3/Class II interaction by LAG-3 D1-D4 Ig was detected (FIG. 5A). However, the LAG-3/MHC Class II interaction can be partially and non-specifically inhibited by human CD8 Ig and IgG1. Moreover, LAG-3 Ig proved to be a potential inhibitor of CD4/Class II interaction (FIG. 5B) under experimental conditions in which CD4/MHC Class II interaction was not modified by human CD8 Ig or IgG1. This suggests that LAG-3/Class II interaction is weaker than CD4/Class II interaction. These results represent the first proof of a possible competition of soluble molecules in an interaction of MHC Class II with its ligands. 
     EXAMPLE 6 
     Immunosuppressant activity of LAG-3 D1-D4 Ig 
     Functional tests were performed using the proliferation tests described above for the biological activity of the anti-LAG-3 monoclonal antibodies. 
     3 days and 5 days (D3 and D5) after antigenic stimulation, LAG-3 D1-D4 Ig showed a strong inhibition of the proliferation of clone 28, while human CD8 Ig and IgG had no effect (FIG. 6). Similar experiments were carried out with clone 154 (FIG. 7), and showed a partial inhibition in the presence of LAG-3 Ig. A control carried out with anti-LAG-3 monoclonal antibodies had the reverse effects, as observed previously. 
     A significant inhibition of the cell proliferation of cells incubated in the presence of LAG-3 D1-D4 Ig was also observed for clone 28. 
     These observations show that LAG-3 D1-D4 Ig is a potential immunosuppressant of the proliferation of T lymphocytes stimulated by an antigen, and indicate that LAG-3 might act as an &#34;extinguisher&#34; of the secondary immune response induced by activated CD4 +   T helper lymphocytes. 
     Role of LAG-3Ig in the negative regulation of the immune response of T cells 
     To demonstrate that a soluble form of LAG-3, mimicking the functions of the membrane molecule, could inhibit the activation of CD 4   +   T clones stimulated by an antigen, the following tests were carried out on clone T154: the T cells are incubated beforehand with a saturating amount of LAG-3Ig (100 nM). The cells are then washed twice with cold RPMI and incubated with 10 μg/ml of goat antibodies directed against human immunoglobulins (Tago) at 4° C. for 30 minutes. 
     After two more washes, the cells are resuspended in RPMI containing 10% of foetal calf serum and incubated for 2 hours at 37° C. before adding the signal. To couple (&#34;cross-link&#34;) the monoclonal antibodies, a goat anti-mouse antibody at a concentration of 10 μg/ml (Tago) is used. 
     FIG. 11 depicts an experiment in which clone T154 has been preincubated with LAG-3Ig bound (&#34;cross-linked&#34;) to a second reactant (polyclonal serum specific for the constant region of human immunoglobulins). The degree of binding of LAG-3Ig to the cells is measured by immunofluorescence (FIG. 11A). FIG. 11B shows that a more than 50% inhibition of the proliferation of clone T154 is produced by LAG-3Ig. Under the same experimental conditions, no effect is observed with the control CD8Ig or with LAG-3Ig without &#34;cross-linking&#34; (not shown in the figure). 
     FIG. 11C also shows that no effect is observed when LAG-3Ig is used to bind (&#34;cross-link&#34;) the MHC Class II molecules expressed by antigen-presenting B cells. 
     The possible effects of bound (&#34;cross-linked&#34;) anti-Class II monoclonal antibodies in relation to the proliferation of T cells were compared to those of LAG-3Ig. A weak inhibition (less than 50% is observed with antibody 949 and antibody D1.12 (anti-DR) bound to a goat anti-mouse polyclonal serum (FIG. 12). The inhibition of proliferation is hence epitope-dependent, the largest effect being obtained with the epitope of LAG-3 specific for the binding to Classes II. 
     The effects of LAG-3Ig on the proliferation of T cells were also studied using different signals on another CD 4   +   T clone, clone TDEL specific for peptide 34-53 of the basic myelin protein. 
     An inhibition of proliferation is observed (n=2) when TDEL is stimulated with the antigen (not shown), with immobilized OKT3 (FIG. 13A), with lectins (PHA+PMA) (FIG. 13B) and with 5 IU/ml of IL 2  (FIG. 13C). No inhibition is observed with 100 IU/ml of IL 2  (FIG. 13D). 
     In conclusion, these results collectively show that LAG-3 and MHC Class II molecules, which are each T cell-activating antigens, may be likened to effector molecules involved in the phase of inactivation of T cell responses. Moreover, these results illustrate the importance of interactions between T cells in the control of the cellular immune response. 
     EXAMPLE 7 
     Stimulation of cell cytotoxicity by LAG-3Ig 
     The role of LAG-3Ig in relation to cell cytotoxicity is studied on two types of effector cells: 
     freshly drawn human peripheral blood lymphocytes (PBL), 
     S1B5 line cells (clone of human NK cells). 
     The cytotoxic activity of these cells is measured by counting the  51  Cr released into the medium by previously labelled target cells, in the presence or absence of LAG-3Ig in the medium. 
     FIG. 14 shows the degree of cytotoxicity of S1B5 for a line of human B cells transformed by Epstein-Barr virus and carrying major histocompatibility complex Class I and II antigens (LAB 388 line), as a function of different reactants added to the cultures. 
     Measurements are carried out after 4 hours of coculture for effector/target (S1B5/LAZ 388) cell ratios of 3:1 (clear columns) of 1:1 (shaded columns). 
     The negative controls consist of medium alone (MED), the immunoadhesin CD8Ig and the monoclonal antibody 17.B4 (anti-LAG-3). 
     The positive controls consist of three different monoclonal antibodies: 
     antibody L243 directed against Class II DR antigens, 
     antibody 9.49 directed against Class II DR, DP DQ antigens, 
     antibody W632 directed against human major histocompatibility complex Class I antigens. 
     Anti-HLA Class I (W632) or Class II (L243) antibodies increase the lysis of the target cells (and not the 17B4 control). The immunoadhesin LAG-3Ig increases the lysis. The CD8Ig control has no effect. 
     FIG. 15 shows the results of an experiment similar to the above, in which the cytotoxicity of PBL with respect to Daudi cells (HLA Class I) is measured, for effector/target ratios of 50:1 (clear columns) and 15:1 (shaded columns). The reactants added to the medium are the same as the ones used in the first experiment, except for antibody 9.49 and antibody 17.B4. Antibody 10H3 is an isotype IgG1 immunoglobulin specific for the CD45 surface antigen. It is used as negative control. 
     No change is observed with an antibody directed against major histocompatibility complex Class I antigens (W632). 
     The data from these two series of measurements show that, compared to negative controls, LAG-3Ig activates the cytotoxicity of NK cells. This effect is similar to the one observed antibodies directed against MHC Class II molecules. 
     BIBLIOGRAPHIC REFERENCES 
     1. TRIEBEL T. et al., 1990, J. Exp. Med. 171, 1393-1405 
     2. BAIXERAS E. et al., 1992, J. Exp. Med. 176, 327-337 
     3. COSGROVE D. et al., 1991, Cell 66, 1051-1066 
     4. RAHEMTULLA A. et al., 1991, Nature 353, 180-184 
     5. TRAUNECKER A. et al., 1988, Nature 331, 84-86 
     6. BENEDICT A. A. et al., 1967, Methods in Immunology 1, 197-306 (1967) 
     7. YELTON D. E. et al., Ann. Rev. of Biochem. 50, 657-680 (1981) 
     8. HUARD B. et al., Immunogenetics 39:213 
     9. MANIATIS T. et al. (1982), Molecular cloning: A laboratory manual--Cold Spring Harbor Laboratory, New-York. 
     10. SEED B., 1987, Nature 329, 840-842 
     11. COLE S. C. et al., Biotechnology 11, 1014-1024, 1993 
     12. COLE S. C. et al., Biotechnology 11, 1014-1024, 1993. 
     
                                           TABLE No. 1__________________________________________________________________________                    residue                           atom type,Atom                     name and                           charge andname    x      y      z      no.    no.__________________________________________________________________________N   25.172370911      27.259836197             67.855064392                    AP-n 40                           n3                             -0.5000                                  1HN2 25.667764664      26.471963882             67.420585632                    AP-n 40                           hn                             0.1300                                  2CA  24.625223160      26.867494583             69.180244446                    AP-n 40                           ca                             0.1200                                  3HN1 24.393711090      27.474891663             67.220008850                    AP-n 40                           hn                             0.1300                                  4HA  23.936895370      27.680395126             69.464080811                    AP-n 40                           h 0.0700                                  5C   25.662780762      26.773513794             70.350120544                    AP-n 40                           c&#39;                             0.3800                                  6O   25.295415878      27.090747833             71.482070923                    AP-n 40                           o&#39;                             -0.4100                                  7CB  23.766021729      25.587018967             68.990669250                    AP-n 40                           c2                             -0.2600                                  8HB1 23.060285568      25.727443695             68.152351379                    AP-n 40                           h 0.0700                                  9HB2 24.413969040      24.744903564             68.686981201                    AP-n 40                           h 0.0700                                  10CG  22.921775818      25.153419495             70.196960449                    AP-n 40                           c-                             0.3400                                  11OD1 22.069602966      25.929233551             70.676017761                    AP-n 40                           o-                             -0.5700                                  12OD2 23.115716934      24.009321213             70.667663574                    AP-n 40                           o-                             -0.5700                                  13N   26.906179428      26.304588318             70.124969482                    SER  41                           n -0.5000                                  14CA  27.860145569      25.912786484             71.207519531                    SER  41                           ca                             0.1200                                  15HN  27.120641708      26.221813202             69.126319885                    SER  41                           hn                             0.2800                                  16HA  27.374551773      25.088045120             71.766326904                    SER  41                           h 0.1000                                  17C   28.252065659      27.005065918             72.271789551                    SER  41                           c&#39;                             0.3800                                  18O   27.987834930      28.200620651             72.115295410                    SER  41                           o&#39;                             -0.3800                                  19CB  29.093601227      25.298025131             70.494880676                    SER  41                           c2                             -0.1700                                  20HB1 28.786190033      24.599395752             69.690521240                    SER  41                           h 0.1000                                  21HB2 29.691858292      26.092912674             70.004081726                    SER  41                           h 0.1000                                  22OG  29.905221939      24.578149796             71.424118042                    SER  41                           oh                             -0.3800                                  23HG  30.662555695      24.231645584             70.939277649                    SER  41                           ho                             0.3500                                  24N   28.857526779      26.558052063             73.387054443                    GLY  42                           n -0.5000                                  25CA  29.199718475      27.440891266             74.535232544                    GLY  42                           cg                             0.0200                                  26HN  29.251720428      25.616510391             73.267890930                    GLY  42                           hn                             0.2800                                  27HA1 28.520187378      28.312047958             74.591857910                    GLY  42                           h 0.1000                                  28HA2 28.983697891      26.890144348             75.468444824                    GLY  42                           h 0.1000                                  29C   30.691051483      27.875793657             74.601707458                    GLY  42                           c&#39;                             0.3800                                  30O   31.504199982      27.026502609             74.980445862                    GLY  42                           o&#39;                             -0.3800                                  31N   31.113182068      29.132183075             74.266487122                    PRO  43                           n -0.4200                                  32CA  32.558349609      29.476200104             74.126792908                    PRO  43                           ca                             0.0600                                  33HA  33.096603394      28.605407715             73.708091736                    PRO  43                           h 0.1000                                  34CD  30.214075089      30.174203873             73.722633362                    PRO  43                           c2                             0.0600                                  35HD1 29.467987061      30.516298294             74.466743469                    PRO  43                           h 0.1000                                  36HD2 29.664882660      29.799777985             72.838768005                    PRO  43                           h 0.1000                                  37C   33.318023682      29.988374710             75.414916992                    PRO  43                           c&#39;                             0.3800                                  38O   32.682361603      30.557033539             76.312332153                    PRO  43                           o&#39;                             -0.3800                                  39CB  32.483139038      30.574260712             73.043136597                    PRO  43                           c2                             -0.2000                                  40HB1 33.350620270      31.263189316             73.049049377                    PRO  43                           h 0.1000                                  41HB2 32.463790894      30.110534568             72.036743164                    PRO  43                           h 0.1000                                  42CG  31.160949707      31.299722672             73.307487488                    PRO  43                           c2                             -0.2000                                  43HG1 31.279897690      32.024162292             74.137779236                    PRO  43                           h 0.1000                                  44HG2 30.794561386      31.862808228             72.428352356                    PRO  43                           h 0.1000                                  45N   34.683673859      29.902477264             75.503486633                    PRO  44                           n -0.4200                                  46CA  35.485736847      30.679145813             76.490524292                    PRO  44                           ca                             0.0600                                  47HA  35.018527985      30.645456314             77.491592407                    PRO  44                           h 0.1000                                  48CD  35.509281158      29.014368057             74.655700684                    PRO  44                           c2                             0.0600                                  49HD1 35.411357880      29.247959137             73.577461243                    PRO  44                           h 0.1000                                  50HD2 35.214973450      27.955932617             74.801994324                    PRO  44                           h 0.1000                                  51C   35.700843811      32.172924042             76.063224792                    PRO  44                           c&#39;                             0.3800                                  52O   36.448230743      32.477428436             75.126922607                    PRO  44                           o&#39;                             -0.3800                                  53CB  36.779544830      29.842718124             76.547348022                    PRO  44                           c2                             -0.2000                                  54HB1 37.662769318      30.430650711             76.863670349                    PRO  44                           h 0.1000                                  55HB2 36.667564392      29.027103424             77.288825989                    PRO  44                           h 0.1000                                  56CG  36.940769196      29.250995636             75.143180847                    PRO  44                           c2                             -0.2000                                  57HG1 37.446662903      29.982709886             74.483322144                    PRO  44                           h 0.1000                                  58HG2 37.553295135      28.329860687             75.134750366                    PRO  44                           h 0.1000                                  59N   35.026676178      33.104183197             76.753837585                    ALA  45                           n -0.5000                                  60CA  35.034278870      34.544979095             76.400695801                    ALA  45                           ca                             0.1200                                  61HN  34.452354431      32.747509003             77.536170959                    ALA  45                           hn                             0.2800                                  62HA  35.105010986      34.659946442             75.298950195                    ALA  45                           h 0.1000                                  63C   36.209384918      35.322113037             77.076705933                    ALA  45                           c&#39;                             0.3800                                  64O   36.163528442      35.637268066             78.268325806                    ALA  45                           o&#39;                             -0.3800                                  65CB  33.646369934      35.083190918             76.800727844                    ALA  45                           c3                             -0.3000                                  66HB1 33.534698486      36.150661469             76.535202026                    ALA  45                           h 0.1000                                  67HB2 32.628392029      34.539138794             76.290328979                    ALA  45                           h 0.1000                                  68HB3 33.465579987      35.001335144             77.890525818                    ALA  45                           h 0.1000                                  69N   37.266757965      35.613758087             76.297294617                    ALA  46                           n -0.5000                                  70HN  37.216701508      35.231555939             75.346885681                    ALA  46                           hn                             0.2800                                  71CA  38.489383698      36.310386658             76.786270142                    ALA  46                           ca                             0.1200                                  72HA  38.262126923      36.871456146             77.716934204                    ALA  46                           h 0.1000                                  73C   39.058414459      37.311935425             75.727844238                    ALA  46                           c&#39;                             0.3800                                  74O   38.922710419      37.100418091             74.516731262                    ALA  46                           o&#39;                             -0.3800                                  75CB  39.526046753      35.215301514             77.108406067                    ALA  46                           c3                             -0.3000                                  76HB1 40.446556091      35.633434296             77.555480957                    ALA  46                           h 0.1000                                  77HB2 39.131206512      34.469978333             77.822120667                    ALA  46                           h 0.1000                                  78HB3 39.821903229      34.663463593             76.197715759                    ALA  46                           h 0.1000                                  79N   39.737388611      38.384365082             76.180320740                    ALA  47                           n -0.5000                                  80CA  40.295833588      39.429889679             75.275863647                    ALA  47                           ca                             0.1200                                  81HN  39.717037201      38.512592316             77.196357727                    ALA  47                           hn                             0.2800                                  82HA  39.901103973      39.319335938             74.245994568                    ALA  47                           h 0.1000                                  83C   41.869789124      39.413467407             75.170806885                    ALA  47                           c&#39;                             0.3800                                  84O   42.518333435      40.166862488             75.906578064                    ALA  47                           o&#39;                             -0.3800                                  85CB  39.722030640      40.769996643             75.786285400                    ALA  47                           c3                             -0.3000                                  86HB1 40.045078278      41.611721039             75.145561218                    ALA  47                           h 0.1000                                  87HB2 38.615306854      40.778987885             75.787467957                    ALA  47                           h 0.1000                                  88HB3 40.059757233      41.007873535             76.813346863                    ALA  47                           h 0.1000                                  89N   42.537422180      38.621597290             74.274406433                    PRO  48                           n -0.4200                                  90CA  44.009857178      38.718185425             74.045745850                    PRO  48                           ca                             0.0600                                  91HA  44.539390564      38.855972290             75.008773804                    PRO  48                           h 0.1000                                  92CD  41.903011322      37.522361755             73.516281128                    PRO  48                           c2                             0.0600                                  93HD1 41.081176758      37.871139526             72.860626221                    PRO  48                           h 0.1000                                  94HD2 41.490711212      36.761222839             74.206848145                    PRO  48                           h 0.1000                                  95C   44.448692322      39.844142914             73.044319153                    PRO  48                           c&#39;                             0.3800                                  96O   43.693031311      40.225761414             72.144134521                    PRO  48                           o&#39;                             -0.3800                                  97CB  44.301902771      37.304885864             73.500595093                    PRO  48                           c2                             -0.2000                                  98HB1 45.227554321      37.245040894             72.897834778                    PRO  48                           h 0.1000                                  99HB2 44.442562103      36.597515106             74.341529846                    PRO  48                           h 0.1000                                  100CG  43.051033020      36.925685883             72.702011108                    PRO  48                           c2                             -0.2000                                  101HG1 43.084365845      37.389282227             71.696128845                    PRO  48                           h 0.1000                                  102HG2 42.948661804      35.835418701             72.555740356                    PRO  48                           h 0.1000                                  103N   45.700798035      40.324272156             73.165817261                    GLY  49                           n -0.5000                                  104CA  46.289730072      41.291931152             72.184875488                    GLY  49                           og                             0.0200                                  105HN  46.207077026      39.986907959             73.991729736                    GLY  49                           hn                             0.2800                                  106HA1 45.620616913      41.460151672             71.317451477                    GLY  49                           h 0.1000                                  107HA2 46.357063293      42.283206940             72.672386169                    GLY  49                           h 0.1000                                  108C   47.682319641      40.950855255             71.600997925                    GLY  49                           c&#39;                             0.3800                                  109O   48.560806274      41.811416626             71.601501465                    GLY  49                           o&#39;                             -0.3800                                  110N   47.842975616      39.718383789             71.091018677                    HIS  50                           n -0.5000                                  111HN  46.947166443      39.222202301             71.052452087                    HIS  50                           hn                             0.2800                                  112CA  49.020114899      39.216835022             70.306198120                    HIS  50                           ca                             0.1200                                  113HA  49.324539185      38.296161652             70.836235046                    HIS  50                           h 0.1000                                  114C   50.400863647      39.993576050             70.182189941                    HIS  50                           c&#39;                             0.3800                                  115O   50.733478546      40.446022034             69.081466675                    HIS  50                           o&#39;                             -0.3800                                  116CB  48.455345154      38.673969269             68.953773499                    HIS  50                           c2                             -0.2000                                  117HB1 49.238502502      38.051708221             68.481765747                    HIS  50                           h 0.1000                                  118HB2 47.639427185      37.951225281             69.144165039                    HIS  50                           h 0.1000                                  119CG  47.954322815      39.695293427             67.919425964                    HIS  50                           c5                             0.1000                                  120ND1 46.759342194      40.404747009             68.035293579                    HIS  50                           np                             -0.4200                                  121CE1 46.825572968      41.133701324             66.873245239                    HIS  50                           c5                             0.2700                                  122NE2 47.887611389      40.964004517             66.014495850                    HIS  50                           np                             -0.5000                                  123CD2 48.617565155      40.019775391             66.717041016                    HIS  50                           c5                             0.0100                                  124HE1 46.043247223      41.852840424             66.655242920                    HIS  50                           h 0.1300                                  125HE2 48.092224121      41.403381348             65.108955383                    HIS  50                           hn                             0.2800                                  126HD2 49.566501617      39.599807739             66.396347046                    HIS  50                           h 0.1300                                  127N   51.278770447      40.105480194             71.226615906                    PRO  51                           n -0.4200                                  128CA  52.667034149      40.618915558             71.077911377                    PRO  51                           ca                             0.0600                                  129HA  52.723186493      41.393623352             70.287094116                    PRO  51                           h 0.1000                                  130CD  50.956447601      39.767082214             72.624496460                    PRO  51                           c2                             0.0600                                  131HD1 50.920970917      38.670558929             72.747383118                    PRO  51                           h 0.1000                                  132HD2 49.988422394      40.190521240             72.947166443                    PRO  51                           h 0.1000                                  133C   53.707103729      39.478122711             70.804489136                    PRO  51                           c&#39;                             0.3800                                  134O   53.623699188      38.391666412             71.391418457                    PRO  51                           o&#39;                             -0.3800                                  135CB  52.846694946      41.283794403             72.455955505                    PRO  51                           c2                             -0.2000                                  136HB1 53.907955170      41.442169189             72.729286194                    PRO  51                           h 0.1000                                  137HB2 52.373264313      42.285846710             72.449127197                    PRO  51                           h 0.1000                                  138CG  52.105823517      40.370025635             73.440399170                    PRO  51                           c2                             -0.2000                                  139HG1 52.782051086      39.565395355             73.789718628                    PRO  51                           h 0.1000                                  140HG2 51.753723145      40.912036896             74.337333679                    PRO  51                           h 0.1000                                  141N   54.704883575      39.729522705             69.933471680                    LEU  52                           n -0.5000                                  142CA  55.791530609      38.746330261             69.603820801                    LEU  52                           ca                             0.1200                                  143HN  54.653743744      40.652229309             69.490356445                    LEU  52                           hn                             0.2800                                  144HA  56.479202271      39.288208008             68.927917480                    LEU  52                           h 0.1000                                  145C   55.301837921      37.525024414             68.745040894                    LEU  52                           c&#39;                             0.3800                                  146O   55.637695313      37.425930023             67.562049866                    LEU  52                           o&#39;                             -0.3800                                  147CB  56.671585083      38.316761017             70.829437256                    LEU  52                           c2                             -0.2000                                  148HB1 56.036743164      37.710464478             71.502593994                    LEU  52                           h 0.1000                                  149HB2 57.445541382      37.602729797             70.487434387                    LEU  52                           h 0.1000                                  150CG  57.363307953      39.420749664             71.675102234                    LEU  52                           cl                             -0.1000                                  151HG  56.617557526      40.200679779             71.926834106                    LEU  52                           h 0.1000                                  152CD1 57.875057220      38.833915710             73.004547119                    LEU  52                           c3                             -0.3000                                  153HD11    58.353130341      39.601875305             73.642135620                    LEU  52                           h 0.1000                                  154HD12    57.048751831      38.403957367             73.601577759                    LEU  52                           h 0.1000                                  155HD13    58.618564606      38.028480530             72.852462769                    LEU  52                           h 0.1000                                  156CD2 58.531608582      40.085853577             70.927200317                    LEU  52                           c3                             -0.3000                                  157HD21    59.028976440      40.857166290             71.545303345                    LEU  52                           h 0.1000                                  158HD22    59.309509277      39.355545044             70.634819031                    LEU  52                           h 0.1000                                  159HD23    58.192760468      40.592193604             70.005569458                    LEU  52                           h 0.1000                                  160N   54.534648895      36.601863861             69.354270935                    ALA  53                           n -0.5000                                  161CA  53.940563202      35.420303345             68.673507690                    ALA  53                           ca                             0.1200                                  162HN  54.159572601      36.958141327             70.246543884                    ALA  53                           hn                             0.2800                                  163HA  53.600482941      35.753322601             67.671340942                    ALA  53                           h 0.1000                                  164C   52.639778137      34.883575439             69.383995056                    ALA  53                           c&#39;                             0.3800                                  165O   51.628326416      34.818698883             68.677047729                    ALA  53                           o&#39;                             -0.3800                                  166CB  55.008785248      34.330768585             68.423698425                    ALA  53                           c3                             -0.3000                                  167HB1 54.582756042      33.460170746             67.892036438                    ALA  53                           h 0.1000                                  168HB2 55.828662872      34.711517334             67.787384851                    ALA  53                           h 0.1000                                  169HB3 55.479701996      33.961406708             69.351325989                    ALA  53                           h 0.1000                                  170N   52.555103302      34.484893799             70.698265076                    PRO  54                           n -0.4200                                  171CA  51.304950714      33.917499542             71.286506653                    PRO  54                           ca                             0.0600                                  172HA  50.878768921      33.172523499             70.584587097                    PRO  54                           h 0.1000                                  173CD  53.705833435      34.435420990             71.626182556                    PRO  54                           c2                             0.0600                                  174HD1 54.215991974      35.409980774             71.739936829                    PRO  54                           h 0.1000                                  175HD2 54.453365326      33.693523407             71.288909912                    PRO  54                           h 0.1000                                  176C   50.177021027      34.945980072             71.642837524                    PRO  54                           c&#39;                             0.3800                                  177O   50.377983093      36.164409637             71.677795410                    PRO  54                           o&#39;                             -0.3800                                  178CB  51.867893219      33.173828125             72.519187927                    PRO  54                           c2                             -0.2000                                  179HB1 51.135505676      33.051830292             73.340660095                    PRO  54                           h 0.1000                                  180HB2 52.181377411      32.150829315             72.232276917                    PRO  54                           h 0.1000                                  181CG  53.087123871      33.989192963             72.949317932                    PRO  54                           c2                             -0.2000                                  182HG1 52.768703461      34.871643066             73.538475037                    PRO  54                           h 0.1000                                  183HG2 53.791828156      33.414070129             73.579086304                    PRO  54                           h 0.1000                                  184N   48.977436066      34.414421082             71.936706543                    GLY  55                           n -0.5000                                  185CA  47.822620392      35.225021362             72.404747009                    GLY  55                           cg                             0.0200                                  186HN  48.958133698      33.389709473             71.929512024                    GLY  55                           hn                             0.2800                                  187HA1 47.830284119      36.238574982             71.963676453                    GLY  55                           h 0.1000                                  188HA2 46.896526337      34.772380829             72.004432678                    GLY  55                           h 0.1000                                  189C   47.670829773      35.263648987             73.950416565                    GLY  55                           c&#39;                             0.3800                                  190O   47.247509003      34.242198944             74.498336792                    GLY  55                           o&#39;                             -0.3800                                  191N   47.956153870      36.372848511             74.696281433                    PRO  56                           n -0.4200`                                  192CA  47.830066681      36.396587372             76.179130554                    PRO  56                           ca                             0.0600                                  193HA  48.225147247      35.457695007             76.619560242                    PRO  56                           h 0.1000                                  194CD  48.653686523      37.556163788             74.163940430                    PRO  56                           c2                             0.0600                                  195HD1 48.108860016      38.040233612             73.332565308                    PRO  56                           h 0.1000                                  196HD2 49.652721405      37.256084442             73.799308777                    PRO  56                           h 0.1000                                  197C   46.361907959      36.604877472             76.668052673                    PRO  56                           c&#39;                             0.3800                                  198O   45.890090942      37.732730865             76.845039368                    PRO  56                           o&#39;                             -0.3800                                  199CB  48.804897308      37.531764984             76.560951233                    PRO  56                           c2                             -0.2000                                  200HB1 48.542221069      38.034294128             77.511970520                    PRO  56                           h 0.1000                                  201HB2 49.825592041      37.124542236             76.697402954                    PRO  56                           h 0.1000                                  202CG  48.782566071      38.488700867             75.368530273                    PRO  56                           c2                             -0.2000                                  203HG1 47.903289795      39.158111572             75.434867859                    PRO  56                           h 0.1000                                  204HG2 49.679012299      39.133480072             75.321792603                    PRO  56                           h 0.1000                                  205N   45.650573730      35.488880157             76.896896362                    HIS  57                           n -0.5000                                  206HN  46.046119690      34.657379150             76.439048767                    HIS  57                           hn                             0.2800                                  207CA  44.244419098      35.504474640             77.387596130                    HIS  57                           ca                             0.1200                                  208HA  43.667560577      36.207649231             76.759368896                    HIS  57                           h 0.1000                                  209C   44.173530579      35.943927765             78.901000977                    HIS  57                           c&#39;                             0.3800                                  210O   44.750030518      35.234142303             79.736030579                    HIS  57                           o&#39;                             -0.3800                                  211CB  43.610416412      34.095542908             77.185058594                    HIS  57                           c2                             -0.2000                                  212HB1 44.270858765      33.323795319             77.623748779                    HIS  57                           h 0.1000                                  213HB2 42.689029694      34.034877777             77.796188354                    HIS  57                           h 0.1000                                  214CG  43.250400543      33.671833038             75.751731873                    HIS  57                           c5                             0.1000                                  215ND1 44.116428375      33.723644257             74.666069031                    HIS  57                           np                             -0.4200                                  216CE1 43.325973511      33.139041901             73.713264465                    HIS  57                           c5                             0.2700                                  217NE2 42.066158295      32.716590881             74.036811829                    HIS  57                           np                             -0.5000                                  218CD2 42.045005798      33.048488617             75.379264832                    HIS  57                           c5                             0.0100                                  219HE1 43.712070465      33.001682281             72.711807251                    HIS  57                           h 0.1300                                  220HE2 41.370864868      32.225383759             73.464263916                    HIS  57                           hn                             0.2800                                  221HD2 41.236827850      32.815635681             76.058380127                    HIS  57                           h 0.1300                                  222N   43.513858795      37.068405151             79.318038940                    PRO  58                           n -0.4200                                  223CA  43.593978882      37.569736481             80.719970703                    PRO  58                           ca                             0.0600                                  224HA  44.657642365      37.630668640             81.026939392                    PRO  58                           h 0.1000                                  225CD  42.833599091      38.010932922             78.407653809                    PRO  58                           c2                             0.0600                                  226HD1 42.093353271      37.516807556             77.751007080                    PRO  58                           h 0.1000                                  227HD2 43.569110870      38.527889252             77.758674622                    PRO  58                           h 0.1000                                  228C   42.805988312      36.695770264             81.747009277                    PRO  58                           c&#39;                             0.3800                                  229O   41.569808960      36.693984985             81.769706726                    PRO  58                           o&#39;                             -0.3800                                  230CB  43.072513580      39.016571045             80.579101563                    PRO  58                           c2                             -0.2000                                  231HB1 42.559799194      39.392253876             81.485702515                    PRO  58                           h 0.1000                                  232HB2 43.920654297      39.706352234             80.398040771                    PRO  58                           h 0.1000                                  233CG  42.156440735      38.999496460             79.353309631                    PRO  58                           c2                             -0.2000                                  234HG1 41.151851654      38.629653931             79.634307861                    PRO  58                           h 0.1000                                  235HG2 42.023620605      39.998752594             78.896965027                    PRO  58                           h 0.1000                                  236N   43.540618896      35.961261749             82.605430603                    ALA  59                           n -0.5000                                  237HN  44.537330627      35.932937622             82.365699768                    ALA  59                           hn                             0.2800                                  238CA  42.949653625      34.946206954             83.526855469                    ALA  59                           ca                             0.1200                                  239HA  42.197692871      34.355514526             82.965911865                    ALA  59                           h 0.1000                                  240C   42.208984375      35.488863972             84.803985596                    ALA  59                           c&#39;                             0.3800                                  241O   42.496433258      35.127353668             85.948333740                    ALA  59                           o&#39;                             -0.3800                                  242CB  44.107444763      33.975612640             83.839767456                    ALA  59                           c3                             -0.3000                                  243HB1 43.761249542      33.130538940             84.463287354                    ALA  59                           h 0.1000                                  244HB2 44.544620514      33.531585693             82.924507141                    ALA  59                           h 0.1000                                  245HB3 44.925910950      34.467308044             84.399597168                    ALA  59                           h 0.1000                                  246N   41.192062378      36.323741913             84.559051514                    ALA  60                           n -0.5000                                  247HN  41.124549866      36.554912676             83.555343628                    ALA  60                           hn                             0.2800                                  248CA  40.203086853      36.790748596             85.562988281                    ALA  60                           ca                             0.1200                                  249HA  40.023948669      35.965785980             86.283721924                    ALA  60                           h 0.1000                                  250C   38.823528290      37.027305603             84.846977234                    ALA  60                           c&#39;                             0.3800                                  251O   37.897804260      36.283885956             85.186340332                    ALA  60                           o&#39;                             -0.3800                                  252CB  40.756233215      37.971691132             86.389617920                    ALA  60                           c3                             -0.3000                                  253HB1 40.004146576      38.357006073             87.102043152                    ALA  60                           h 0.1000                                  254HB2 41.631908417      37.656120300             86.987319946                    ALA  60                           h 0.1000                                  255HB3 41.090072632      38.818138123             85.765472412                    ALA  60                           h 0.1000                                  256N   38.616340637      37.921970367             83.823921204                    PRO  61                           n -0.4200                                  257CA  37.403762817      37.874496460             82.948486328                    PRO  61                           ca                             0.0600                                  258HA  36.491020203      37.824863434             83.573593140                    PRO  61                           h 0.1000                                  259CD  39.556003571      39.003845215             83.459777832                    PRO  61                           c2                             0.0600                                  260HD1 40.594814301      38.658184052             83.304046631                    PRO  61                           h 0.1000                                  261HD2 39.571029663      39.778694153             84.250648499                    PRO  61                           h 0.1000                                  262C   37.275394440      36.712963104             81.892074585                    PRO  61                           c&#39;                             0.3800                                  263O   36.266387939      36.670307159             81.183494568                    PRO  61                           o&#39;                             -0.3800                                  264CB  37.473857880      39.266963959             82.286987305                    PRO  61                           c2                             -0.2000                                  265HB1 36.949714661      39.321308136             81.312759399                    PRO  61                           h 0.1000                                  266HB2 36.985511780      40.017913818             82.938987732                    PRO  61                           h 0.1000                                  267CG  38.968631744      39.571613312             82.168632507                    PRO  61                           c2                             -0.2000                                  268HG1 39.397396088      39.046298981             81.292793274                    PRO  61                           h 0.1000                                  269HG2 39.180202484      40.649600983             82.039207458                    PRO  61                           h 0.1000                                  270N   38.242324829      35.777751923             81.785034180                    SER  62                           n -0.5000                                  271CA  38.186004639      34.624855042             80.844123840                    SER  62                           ca                             0.1200                                  272HN  39.035541534      35.927360535             82.416992188                    SER  62                           hn                             0.2800                                  273HA  37.921787262      35.006183624             79.841125488                    SER  62                           h 0.1000                                  274C   37.145660400      33.532897949             81.261909485                    SER  62                           c&#39;                             0.3800                                  275O   37.466091156      32.626064301             82.041137695                    SER  62                           c&#39;                             -0.3800                                  276CB  39.620265961      34.048240662             80.725196838                    SER  62                           c2                             -0.1700                                  277HB1 39.660293579      33.306644440             79.904281616                    SER  62                           h 0.1000                                  278HB2 40.349685669      34.832687378             88.442802429                    SER  62                           h 0.1000                                  279OG  40.032703400      33.414188385             81.938880920                    SER  62                           oh                             -0.3800                                  280HG  39.252223969      32.931293488             82.256263733                    SER  62                           ho                             0.3500                                  281N   35.902244568      33.647918701             80.764541626                    SER  63                           n -0.5000                                  282CA  34.747528076      32.874465942             81.297317505                    SER  63                           ca                             0.1200                                  283HN  35.768447876      34.503852844             80.205200195                    SER  63                           hn                             0.2800                                  284HA  35.064254761      32.265518188             82.170570374                    SER  63                           h 0.1000                                  215C   34.106758118      31.936998367             80.231674194                    SER  63                           c&#39;                             0.3800                                  286O   33.716896057      32.367130280             79.142120361                    SER  63                           o&#39;                             -0.3800                                  287CB  33.716815948      33.889484406             81.843544006                    SER  63                           c2                             -0.1700                                  288HB1 34.199871063      34.571384430             82.572105408                    SER  63                           h 0.1000                                  289HB2 33.328830719      34.543502808             81.036796570                    SER  63                           h 0.1000                                  290OG  32.634590149      33.222091675             82.496467590                    SER  63                           oh                             -0.3800                                  291HG  32.159793854      32.710407257             81.832328796                    SER  63                           ho                             0.3500                                  292N   33.914913177      30.658897400             80.588378906                    TRP  64                           n -0.5000                                  293CA  33.112319946      29.694124222             79.783546448                    TRP  64                           ca                             0.1200                                  294HN  34.221500397      30.422899246             81.538955688                    TRP  64                           hn                             0.2800                                  295HA  33.404731750      29.812168121             78.721931458                    TRP  64                           h 0.1000                                  296C   31.573041916      29.961977005             79.883049011                    TRP  64                           c&#39;                             0.3800                                  297O   30.996980667      29.940891266             80.975959778                    TRP  64                           o&#39;                             -0.3800                                  298CB  33.525466919      28.235471725             80.142707825                    TRP  64                           c2                             -0.2000                                  299HB1 32.950366974      27.534402847             79.508041382                    TRP  64                           h 0.1000                                  300HB2 34.571674347      28.078489304             79.816658020                    TRP  64                           h 0.1000                                  301CG  33.405326843      27.783784866             81.611763000                    TRP  64                           c5                             0.0000                                  302CD1 32.267101288      27.214570999             82.221214294                    TRP  64                           c5                             0.0100                                  303NE1 32.481933594      26.953943253             83.590408325                    TRP  64                           np                             -0.5000                                  304CE2 33.781982422      27.378627777             83.812339783                    TRP  64                           c5                             0.1100                                  305CD2 34.355152130      27.881061554             82.617965698                    TRP  64                           c5                             0.0000                                  306HD1 31.332025528      27.036033630             81.708480835                    TRP  64                           h 0.1000                                  307HE1 31.820940018      26.578481674             84.279945374                    TRP  64                           hn                             0.2800                                  308CE3 35.681034088      28.394184113             82.615905762                    TRP  64                           cp                             -0.1000                                  309HE3 36.128986359      28.784986496             81.714393616                    TRP  64                           h 0.1000                                  310CZ3 36.396430969      28.387191772             83.815704346                    TRP  64                           cp                             -0.1000                                  311HZ3 37.405311584      28.776082993             83.832160950                    TRP  64                           h 0.1000                                  312CH2 35.830604553      27.888952255             84.994010925                    TRP  64                           cp                             -0.1000`                                  313HH2 36.410472870      27.896844864             85.906951904                    TRP  64                           h 0.1000                                  314CZ2 34.527233124      27.382776260             85.014289856                    TRP  64                           cp                             -0.1000                                  315HZ2 34.097515106      27.006088257             85.929389954                    TRP  64                           h 0.1000                                  316N   30.921329498      30.232547760             78.740600586                    GLY  65                           n -0.5000                                  317CA  29.460748672      30.504768372             78.692192078                    GLY  65                           cg                             0.0200                                  318HN  31.520374298      30.302478790             77.901519775                    GLY  65                           hn                             0.2800                                  319HA1 29.073087692      30.896234512             79.650825500                    GLY  65                           h 0.1000                                  320HA2 29.288171768      31.333106995             77.981094360                    GLY  65                           h 0.1000                                  321C   28.633579254      29.293350220             78.197364807                    GLY  65                           c&#39;                             0.3800                                  322O   28.566907883      29.137302399             76.975486755                    GLY  65                           o&#39;                             -0.3800                                  323N   27.989013672      28.429246902             79.038352966                    PRO  66                           n -0.4200                                  324CA  27.282257080      27.212890625             78.546752930                    PRO  66                           ca                             0.0600                                  325HA  27.989650726      26.634012222             77.917152405                    PRO  66                           h 0.1000                                  326CD  28.016592026      28.532529831             80.511337280                    PRO  66                           c2                             0.0600                                  327HD1 27.731332779      29.536006927             80.880874634                    PRO  66                           h 0.1000                                  328HD2 29.027824402      28.301166534             80.897590637                    PRO  66                           h 0.1000                                  329C   25.977466583      27.479314804             77.725341797                    PRO  66                           c&#39;                             0.3800                                  330O   25.217950821      28.417282104             77.982574463                    PRO  66                           o&#39;                             -0.3800                                  331CB  27.045602798      26.422634125             79.851196289                    PRO  66                           c2                             -0.2000                                  332HB1 26.132562637      25.797079086             79.827728271                    PRO  66                           h 0.1000                                  333HB2 27.890687943      25.729280472             80.029365540                    PRO  66                           h 0.1000                                  334CG  27.003501892      27.477243423             80.958015442                    PRO  66                           c2                             -0.2000                                  335HG1 25.990892410      27.921483994             81.014678955                    PRO  66                           h 0.1000                                  336HG2 27.232566833      27.061700821             81.956459045                    PRO  66                           h 0.1000                                  337N   25.734319687      26.626403809             76.719802856                    ARG+ 67                           n -0.5000                                  338CA  24.603988647      26.793025970             75.767257690                    ARG+ 67                           ca                             0.1200                                  339HN  26.386735916      25.841371536             76.649803162                    ARG+ 67                           hn                             0.2800                                  340HA  24.496238708      27.874828339             75.561668396                    ARG+ 67                           h 0.1000                                  341C   23.227464676      26.224872589             76.267372131                    ARG+ 67                           c&#39;                             0.3800                                  342O   23.178310394      25.034952164             76.603759766                    ARG+ 67                           o&#39;                             -0.3800                                  343CB  24.990058899      26.165229797             74.398826599                    ARG+ 67                           c2                             -0.2000                                  344HB1 24.135663986      26.318639755             73.709175110                    ARG+ 67                           h 0.1100                                  345HB2 25.787433624      26.779323578             73.940032959                    ARG+ 67                           h 0.1100                                  346CG  25.439929962      24.676465988             74.361564636                    ARG+ 67                           c2                             -0.2000                                  347HG1 26.546255112      24.646316528             74.415458679                    ARG+ 67                           h 0.1300                                  348HG2 25.092346191      24.131168365             75.261718750                    ARG+ 67                           h 0.1300                                  349CD  24.934387207      23.941221237             73.112297058                    ARG+ 67                           c2                             -0.0900                                  350HD1 23.838283539      23.774566650             73.188652039                    ARG+ 67                           h 0.1300                                  351HD2 25.070211411      24.585262299             72.220893860                    ARG+ 67                           h 0.1300                                  352NE  25.665744781      22.657058716             72.968780518                    ARG+ 67                           n1                             -0.5000                                  353HE  36.251846313      22.313375473             73.731925964                    ARG+ 67                           hn                             0.3600                                  354CZ  25.689014435      21.902687073             71.871635437                    ARG+ 67                           cr                             0.4500                                  355NH1 26.493299484      20.879484177             71.859733582                    ARG+ 67                           n2                             -0.5000                                  356HH11    27.072675705      20.740955353             72.690315247                    ARG+ 67                           hn                             0.3600                                  357HH12    26.520929337      20.319580078             71.006805420                    ARG+ 67                           hn                             0.3600                                  358NH2 24.956668854      22.117029190             70.805320740                    ARG+ 67                           n2                             -0.5000                                  359HH21    25.030595779      21.456792831             70.033142090                    ARG+ 67                           hn                             0.3600                                  360HH22    24.266489029      22.916227341             70.850784302                    ARG+ 67                           hn                             0.3600                                  361N   22.080270767      26.971176147             76.244255066                    PRO  68                           n -0.4200                                  362CA  20.743734360      26.358839035             76.485237122                    PRO  68                           ca                             0.0600                                  363HA  20.817556381      25.605155945             77.294357300                    PRO  68                           h 0.1000                                  364CD  22.076143265      28.448001862             76.342918396                    PRO  68                           c2                             0.0600                                  365HD1 22.539228439      28.949869156             75.469612122                    PRO  68                           h 0.1000                                  366HD2 22.632146835      28.776126862             77.244499207                    PRO  68                           h 0.1000                                  367C   20.182382584      25.586124692             75.240180969                    PRO  68                           c&#39;                             0.3800                                  368O   20.420539856      24.381649017             75.139877319                    PRO  68                           o&#39;                             -0.3800                                  369CB  19.948141098      27.549791336             77.062515259                    PRO  68                           c2                             -0.2000                                  370HB1 18.858430862      27.490163803             76.877128601                    PRO  68                           h 0.1000                                  371HB2 20.066789627      27.567596436             78.163002014                    PRO  68                           h 0.1000                                  372CG  20.592071533      28.804227829             76.467758179                    PRO  68                           c2                             -0.2000                                  373HG1 20.158363342      29.031393051             75.478172302                    PRO  68                           h 0.1000                                  374HG2 20.428398132      29.704229355             77.092338562                    PRO  68                           h 0.1000                                  375N   19.458055496      26.229068756             74.300010681                    ARG+ 69                           n -0.5000                                  376CA  18.893756866      25.542047501             73.096710205                    ARG+ 69                           ca                             0.1200                                  377HN  19.221296310      27.198928833             74.529014587                    ARG+ 69                           hn                             0.2800                                  378HA  19.667610168      24.872461319             72.660797119                    ARG+ 69                           h 0.1000                                  379C   18.514461517      26.608341217             72.012504578                    ARG+ 69                           c&#39;                             0.3800                                  380O   17.383218765      27.099323273             72.036094666                    ARG+ 69                           o&#39;                             -0.3800                                  381CB  17.681777954      24.654710770             73.539657593                    ARG+ 69                           c2                             -0.2000                                  382HB1 18.011884689      23.935596466             74.315147400                    ARG+ 69                           h 0.1100                                  383HB2 16.954420090      25.310214996             74.061965942                    ARG+ 69                           h 0.1100                                  384CG  16.946891785      23.862810135             72.427490234                    ARG+ 69                           c2                             -0.2000                                  385HG1 16.649517059      24.551628113             71.612152100                    ARG+ 69                           h 0.1300                                  386HG2 17.638776779      23.127803802             71.965805054                    ARG+ 69                           h 0.1300                                  387CD  15.688191414      23.166488647             72.975959778                    ARG+ 69                           c2                             -0.0900                                  388HD1 15.980383873      22.365550995             73.686569214                    ARG+ 69                           h 0.1300                                  389HD2 15.090404510      23.898859024             73.554351807                    ARG+ 69                           h 0.1300                                  390NE  14.889810562      22.612804413             71.848815918                    ARG+ 69                           n1                             -0.5000                                  391HE  15.272388458      22.616128922             70.898582458                    ARG+ 69                           hn                             0.3600                                  392CZ  13.644455910      22.143890381             71.942466736                    ARG+ 69                           cr                             0.4500                                  393NH1 13.048615456      21.755460739             70.850708008                    ARG+ 69                           n2                             -0.5000                                  394HH11    13.576630592      21.832328796             69.979103088                    ARG+ 69                           hn                             0.3600                                  395HH12    12.090744019      21.411947250             70.936080933                    ARG+ 69                           hn                             0.3600                                  396NH2 12.989639282      22.056533813             73.074882507                    ARG+ 69                           n2                             -0.5000                                  397HH21    12.033639908      21.696094513             73.066261292                    ARG+ 69                           hn                             0.3600                                  398HH22    13.529273987      22.372974396             73.883934021                    ARG+ 69                           hn                             0.3600                                  399N   19.436628342      26.928932190             71.074501038                    ARG+ 70                           n -0.5000                                  400CA  19.223009109      27.811206818             69.878326416                    ARG+ 70                           ca                             0.1200                                  401HN  20.357131958      26.451332092             71.165985107                    ARG+ 70                           hn                             0.2800                                  402HA  19.087514877      27.065124512             69.071128845                    ARG+ 70                           h 0.1000                                  403C   20.512538910      28.575149536             69.398536682                    ARG+ 70                           c&#39;                             0.3800                                  404O   20.872812271      28.468791962             68.228363037                    ARG+ 70                           o&#39;                             -0.3800                                  405CB  17.935552597      28.697717667             69.732887268                    ARG+ 70                           c2                             -0.2000                                  406HB1 17.889257431      29.085472107             68.694030762                    ARG+ 70                           h 0.1100                                  407HB2 17.053388596      28.033058167             69.807891846                    ARG+ 70                           h 0.1100                                  408CG  17.775762558      29.883768082             70.717842102                    ARG+ 70                           c2                             -0.2000                                  409HG1 18.004568100      29.546030045             71.747383118                    ARG+ 70                           h 0.1300                                  410HG2 18.540782928      30.651863098             70.495643616                    ARG+ 70                           h 0.1300                                  411CD  16.368116379      30.502414703             70.693214417                    ARG+ 70                           c2                             -0.0900                                  412HD1 16.095691681      30.812221527             69.664886475                    ARG+ 70                           h 0.1300                                  413HD2 15.630161285      29.711160660             70.937812805                    ARG+ 70                           h 0.1300                                  414NE  16.255954742      31.590759277             71.711013794                    ARG+ 70                           n1                             -0.5000                                  415HE  16.253637314      31.350564957             72.706428528                    ARG+ 70                           hn                             0.3600                                  416CZ  16.144437790      32.900745392             71.464541462                    ARG+ 70                           cr                             0.4500                                  417NH1 16.055492401      33.712890625             72.481109619                    ARG+ 70                           n2                             -0.5000                                  418HH11    16.071464539      33.294216156             73.433330078                    ARG+ 70                           hn                             0.3600                                  419HH12    15.975571632      34.708621979             72.277374268                    ARG+ 70                           hn                             0.3600                                  420NH2 16.120351791      33.420074463             70.259872437                    ARG+ 70                           n2                             -0.5000                                  421HH21    16.025018692      34.432674408             70.167800903                    ARG+ 70                           hn                             0.3600                                  422HH22    16.187112808      32.736862183             69.505996704                    ARG+ 70                           hn                             0.3600                                  423N   21.115812302      29.562902451             70.071769714                    TYRC 71                           n -0.5000                                  424HN  21.515977859      30.157514572             69.338050842                    TYRC 71                           hn                             0.2800                                  425CA  22.034273148      29.314456940             71.218978882                    TYRC 71                           ca                             0.1200                                  426HA  22.671009064      28.444923401             70.976676941                    TYRC 71                           h 0.1000                                  427C   21.352920532      28.953493118             72.563385010                    TYRC 71                           c&#39;                             0.4100                                  428OXT 20.392858505      29.553161621             73.048652649                    TYRC 71                           o&#39;                             -0.3800                                  429O   21.928325653      27.853042603             73.145797729                    TYRC 71                           oh                             -03800                                  430HO  21.429273605      27.558662415             73.909782410                    TYRC 71                           ho                             0.3500                                  431CB  22.969152451      30.555358887             71.361984253                    TYRC 71                           c2                             -0.2000                                  432HB1 22.368267059      31.487627029             71.355415344                    TYRC 71                           h 0.1000                                  433HB2 23.401992798      30.559690475             72.381835938                    TYRC 71                           h 0.1000                                  434CG  24.144546509      30.666173935             70.352111816                    TYRC 71                           cp                             0.0000                                  435CD1 23.927837372      31.126276016             69.044418335                    TYRC 71                           cp                             -0.1000                                  436HD1 22.944635391      31.424867630             68.717597961                    TYRC 71                           h 0.1000                                  437CE1 24.987041473      31.212265015             68.143028259                    TYRC 71                           cp                             -0.1000                                  438HE1 24.819047928      31.555503845             67.131973267                    TYRC 71                           h 0.1000                                  439CZ  26.273118973      30.861675262             68.542274475                    TYRC 71                           cp                             0.0300                                  440OH  27.314481735      30.957365036             67.652267456                    TYRC 71                           oh                             -0.3800                                  441HH  26.980180740      31.236070633             66.796859741                    TYRC 71                           ho                             0.3500                                  442CE2 26.504697800      30.422637939             69.841377258                    TYRC 71                           cp                             -0.1000                                  443HE2 27.503232956      30.148866653             70.140815735                    TYRC 71                           h 0.1000                                  444CD2 25.447391510      30.325349808             70.743820190                    TYRC 71                           cp                             -0.1000                                  445HD2 25.652494431      29.968608856             71.745338440                    TYRC 71                           h 0.1000                                  430__________________________________________________________________________ 
    
     
                                           TABLE No. 2__________________________________________________________________________                    residue                           atom type,Atom                     name and                           charge andname    x      y      z      no.    no.__________________________________________________________________________N   24.753738403      26.435615540             68.244300842                    CYSn 40                           n3                             -0.5000                                  1CA  24.503000259      26.356292725             69.707687378                    CYSn 40                           ca                             0.1200                                  2HN1 23.861560822      26.429992676             67.734397888                    CYSn 40                           hn                             0.1400                                  3HN2 25.250690460      25.603431424             67.909477234                    CYSn 40                           hn                             0.1400                                  4HA  23.890571594      27.247760773             69.940788269                    CYSn 40                           h 0.1000                                  5C   25.747190475      26.505004883             70.632949829                    CYSn 40                           c&#39;                             0.3800                                  6O   25.611124039      27.204542160             71.634971619                    CYSn 40                           o&#39;                             -0.3800                                  7CB  23.602088928      25.125436783             69.979545593                    CYSn 40                           c2                             -0.3000                                  8HB1 22.555475235      25.378625870             69.716011047                    CYSn 40                           h 0.1000                                  9HB2 23.865217209      24.277284622             69.317871094                    CYSn 40                           h 0.1000                                  10SG  23.613842010      24.466741562             71.679084778                    CYSn 40                           s1                             0.1000                                  11N   26.910152435      25.881416321             70.350471497                    SER  41                           n -0.5000                                  12CA  28.052598953      25.721015930             71.310394287                    SER  41                           ca                             0.1200                                  13HN  26.921674728      25.473436356             69.412322998                    SER  41                           hn                             0.2800                                  14HA  27.761577606      24.886217117             71.978584290                    SER  41                           h 0.1000                                  15C   28.477056503      26.929338455             72.226699829                    SER  41                           c&#39;                             0.3800                                  16O   28.412267685      28.097608566             71.829902649                    SER  41                           o&#39;                             -0.3800                                  17CB  29.257335663      25.212779999             70.480110168                    SER  41                           c2                             -0.1700                                  18HB1 28.957101822      24.401222229             69.786743164                    SER  41                           h 0.1000                                  19HB2 29.657680511      26.030597687             69.845893860                    SER  41                           h 0.1000                                  20OG  30.284814835      24.713315964             71.339111328                    SER  41                           oh                             -0.3800                                  21HG  31.027490616      24.448732376             70.785003662                    SER  41                           ho                             0.3500                                  22N   28.904022217      26.617259979             73.466476440                    GLY  42                           n -0.5000                                  23CA  29.226711273      27.639400482             74.497131348                    GLY  42                           cg                             0.0200                                  24HN  29.140472412      25.626163483             73.587532043                    GLY  42                           hn                             0.2800                                  25HA1 28.567264557      28.519987106             74.394309998                    GLY  42                           h 0.1000                                  26HA2 28.949186325      27.229663849             75.483924866                    GLY  42                           h 0.1000                                  27C   30.728670120      28.040773392             74.587509155                    GLY  42                           c&#39;                             0.3800                                  28O   31.522300720      27.171119690             74.941143494                    GLY  42                           o&#39;                             -0.3800                                  29N   31.175983429      29.296203613             74.282577515                    PRO  43                           n -0.4200                                  30CA  32.627410889      29.612504959             74.140579224                    PRO  43                           ca                             0.0600                                  31HA  33.153442383      28.731788635             73.723632813                    PRO  43                           h 0.1000                                  32CD  30.395356750      30.374078751             73.784042358                    PRO  43                           c2                             0.0600                                  33HD1 29.605636597      30.735929489             74.571166992                    PRO  43                           h 0.1000                                  34HD2 29.683467865      30.026647568             72.928161621                    PRO  43                           h 0.1000                                  35C   33.389945984      30.112844467             75.429031372                    PRO  43                           c&#39;                             0.3800                                  36O   32.754360199      30.681560516             76.327354431                    PRO  43                           o&#39;                             -0.3800                                  37CB  32.565906525      30.710325241             73.057388306                    PRO  43                           c2                             -0.2000                                  38HB1 33.449081421      31.378034592             73.055488586                    PRO  43                           h 0.1000                                  39HB2 32.524932861      30.247560501             72.051818848                    PRO  43                           h 0.1000                                  40CG  31.263490677      31.466632843             73.332626343                    PRO  43                           c2                             -0.2000                                  41HG1 31.413110733      32.204200745             74.146759033                    PRO  43                           h 0.1000                                  42HG2 30.894020081      32.022109985             72.450286865                    PRO  43                           h 0.1000                                  43N   34.754848480      30.015562057             75.523460388                    PRO  44                           n -0.4200                                  44CA  35.553565979      30.763086319             76.536285400                    PRO  44                           ca                             0.0600                                  45HA  35.083564758      30.695350647             77.536567688                    PRO  44                           h 0.1000                                  46CD  35.574893951      29.135835648             74.665214539                    PRO  44                           c2                             0.0600                                  47HD1 35.471595764      29.373666763             73.588935852                    PRO  44                           h 0.1000                                  48HD2 35.281650543      28.076414108             74.807395935                    PRO  44                           h 0.1000                                  49C   35.767509460      32.265411377             76.141113281                    PRO  44                           c&#39;                             0.3800                                  50O   36.544441223      32.599441528             75.238464355                    PRO  44                           o&#39;                             -0.3800                                  51CB  36.849227905      29.927103043             76.567779541                    PRO  44                           c2                             -0.2000                                  52HB1 37.732776642      30.502979279             76.899009705                    PRO  44                           h 0.1000                                  53HB2 36.733722687      29.095364598             77.289833069                    PRO  44                           h 0.1000                                  54CG  37.005489349      29.369808197             75.152618408                    PRO  44                           c2                             -0.2000                                  55HG1 37.502185822      30.119005203             74.504158020                    PRO  44                           h 0.1000                                  56HG2 37.625408173      28.451385498             75.115615645                    PRO  44                           h 0.1000                                  57N   35.047088623      33.173435211             76.826978455                    ALA  45                           n -0.5000                                  58CA  35.011333466      34.609920502             76.449218750                    ALA  45                           ca                             0.1200                                  59HN  34.471405029      32.798248291             77.590728760                    ALA  45                           hn                             0.2800                                  60HA  35.065380096      34.699813843             75.343650818                    ALA  45                           h 0.1000                                  61C   36.187728882      35.414947510             77.090148926                    ALA  45                           c&#39;                             0.3800                                  62O   36.133388519      35.819305420             78.255142212                    ALA  45                           o&#39;                             -0.3800                                  63CB  33.615478516      35.135440826             76.831176758                    ALA  45                           c3                             -0.3000                                  64HB1 33.490375519      36.187480927             76.517280579                    ALA  45                           h 0.1000                                  65HB2 32.811222076      34.556259155             76.338432312                    ALA  45                           h 0.1000                                  66HB3 33.433517456      35.098365784             77.922439575                    ALA  45                           h 0.1000                                  67N   37.264499664      35.613868713             76.306388855                    ALA  46                           n -0.5000                                  68HN  37.248832703      35.072978973             75.433502197                    ALA  46                           hn                             0.2800                                  69CA  38.503662109      36.298694611             76.764076233                    ALA  46                           ca                             0.1200                                  70HA  38.303600311      36.883266449             77.688095093                    ALA  46                           h 0.1000                                  71C   39.082061768      37.273509979             75.687866211                    ALA  46                           c&#39;                             0.3800                                  72O   38.951850891      37.052509308             74.481193542                    ALA  46                           o&#39;                             -0.3800                                  73CB  39.509185791      35.179004669             77.103065491                    ALA  46                           c3                             -0.3000                                  74HB1 40.441535950      35.582756042             77.535072327                    ALA  46                           h 01000                                  75HB2 39.106670380      34.460605621             77.839447021                    ALA  46                           h 0.1000                                  76HB3 39.780502319      34.597728729             76.205062866                    ALA  46                           h 0.1000                                  77N   39.768814087      38.344066620             76.133750916                    ALA  47                           n -0.5000                                  78CA  40.322643280      39.391151428             75.225708008                    ALA  47                           ca                             0.1200                                  79HN  39.783836365      38.455593109             77.149337769                    ALA  47                           hn                             0.2800                                  80HA  39.932807922      39.265201569             74.196365356                    ALA  47                           h 0.1000                                  81C   41.900882721      39.401374817             75.126907349                    ALA  47                           c&#39;                             0.3800                                  82O   42.538444519      40.171451569             75.854652405                    ALA  47                           o&#39;                             -0.3800                                  83CB  39.728843689      40.731719971             75.714279175                    ALA  47                           c3                             -0.3000                                  84HB1 40.043342590      41.567916870             75.059875488                    ALA  47                           h 0.1000                                  85HB2 38.621978760      40.726497650             75.711242676                    ALA  47                           h 0.1000                                  86HB3 40.062076569      40.987442017             76.739311218                    ALA  47                           h 0.1000                                  87N   42.578651428      38.603939056             74.242019653                    PRO  48                           n -0.4200                                  88CA  44.052474976      38.702857971             74.013595581                    PRO  48                           ca                             0.0600                                  89HA  44.576034546      38.850185394             74.977058411                    PRO  48                           h 0.1000                                  90CD  41.956359863      37.474979401             73.520225525                    PRO  48                           c2                             0.0600                                  91HD1 41.114963531      37.788272858             72.872795105                    PRO  48                           h 0.1000                                  92HD2 41.576156616      36.723818915             74.239135742                    PRO  48                           h 0.1000                                  93C   44.492458344      39.820354462             73.002609253                    PRO  48                           c&#39;                             0.3800                                  94O   43.782276154      40.131282806             72.040626526                    PRO  48                           o&#39;                             -0.3800                                  95CB  44.356296539      37.289741516             73.479736328                    PRO  48                           c2                             -0.2000                                  96HB1 45.273612976      37.234390259             72.865592957                    PRO  48                           h 0.1000                                  97HB2 44.513816833      36.591526031             74.332021484                    PRO  48                           h 0.1000                                  98CG  43.102409363      36.884414673             72.700988770                    PRO  48                           c2                             -0.2000                                  99HG1 43.119277954      37.331111908             71.685241699                    PRO  48                           h 0.1000                                  100HG2 43.010280609      35.788948059             72.572326660                    PRO  48                           h 0.1000                                  101N   45.709655762      40.366821289             73.185493469                    GLY  49                           n -0.5000                                  102CA  46.317604065      41.332912445             73.214889526                    GLY  49                           cg                             0.0200                                  103HN  46.169986725      40.089691162             74.058357239                    GLY  49                           hn                             0.2800                                  104HA1 45.654991150      41.537052155             71.351181030                    GLY  49                           h 0.1000                                  105HA2 46.406318665      42.313266754             72.719123840                    GLY  49                           h 0.1000                                  106C   47.710880280      40.963481903             71.654037476                    GLY  49                           c&#39;                             0.3800                                  107O   48.630664825      41.772521973             71.754951477                    GLY  49                           o&#39;                             -0.3800                                  108N   47.830738068      39.763301849             71.063682556                    HIS  50                           n -0.5000                                  109HN  46.918842316      39.310573578             70.943237305                    HIS  50                           hn                             0.2800                                  110CA  49.045799255      39.210880280             70.375061035                    HIS  50                           ca                             0.1200                                  111HA  49.315334320      38.320884705             70.972137451                    HIS  50                           h 0.1000                                  112C   50.433021545      39.981941223             70.267852783                    HIS  50                           c&#39;                             0.3800                                  113O   50.773132324      40.456230164             69.178672791                    HIS  50                           o&#39;                             -0.3800                                  114CB  48.558776855      38.590164185             69.024101257                    HIS  50                           c2                             -0.2000                                  115HB1 49.390335083      36.009521484             68.577232361                    HIS  50                           h 0.1000                                  116HB2 47.792594910      37.817192078             69.227310181                    HIS  50                           h 0.1000                                  117CG  47.997627258      39.545143127             67.956726074                    HIS  50                           c5                             0.1000                                  118ND1 46.669281006      39.956676483             67.918121338                    HIS  50                           np                             -0.4200                                  119CE1 46.730144501      40.789539337             66.829002380                    HIS  50                           c5                             0.2700                                  120NE2 47.911670685      40.950614929             66.152328491                    HIS  50                           np                             -0.5000                                  121CD2 48.729324341      40.126430511             66.904235840                    HIS  50                           c5                             0.0100                                  122HE1 45.843067169      41.327060699             66.517631531                    HIS  50                           h 0.1300                                  123HE2 48.138290405      41.548683167             65.349815369                    HIS  50                           hn                             0.2800                                  124HD2 49.789726257      39.981491089             66.738136292                    HIS  50                           h 0.1300                                  125N   51.307849884      40.071182251             71.317932129                    PRO  51                           n -0.4200                                  126CA  52.692558289      40.596851349             71.184913635                    PRO  51                           ca                             0.0600                                  127HA  52.742668152      41.390510559             70.412712097                    PRO  51                           h 0.1000                                  128CD  50.980678558      39.703777313             72.706970215                    PRO  51                           c2                             0.0600                                  129HD1 50.998199463      38.605384827             72.818397522                    PRO  51                           h 0.1000                                  130HD2 49.987606049      40.071315765             73.019950867                    PRO  51                           h 0.1000                                  131C   53.739063263      39.471630096             70.880722046                    PRO  51                           c&#39;                             0.3800                                  132O   53.708900452      38.394466400             71.488830566                    PRO  51                           o&#39;                             -0.3800                                  133CB  52.868911743      41.240936279             72.572486877                    PRO  51                           c2                             -0.2000                                  134HB1 53.929355621      41.364253998             72.864852905                    PRO  51                           h 0.1000                                  135HB2 52.429229736      42.258647919             72.565872192                    PRO  51                           h 0.1000                                  136CG  52.087848663      40.349472046             73.547019958                    PRO  51                           c2                             -0.2000                                  137HG1 52.750400543      39.566276550             73.963310242                    PRO  51                           h 0.1000                                  138HG2 51.686916351      40.923263550             74.403717041                    PRO  51                           h 0.1000                                  139N   54.676445007      39.726749420             69.946899414                    LEU  52                           n -0.5000                                  140CA  55.768096924      38.764469147             69.570259094                    LEU  52                           ca                             0.1200                                  141HN  54.573589325      40.637012482             69.488586426                    LEU  52                           hn                             0.2800                                  142HA  56.414031982      39.325927734             68.869346619                    LEU  52                           h 0.1000                                  143C   55.281269073      37.540004730             68.718757629                    LEU  52                           c&#39;                             0.3800                                  144O   55.654800415      37.411125183             67.550910950                    LEU  52                           o&#39;                             -0.3800                                  145CB  56.713882446      38.354763031             70.751991272                    LEU  52                           c2                             -0.2000                                  146HB1 56.125553131      37.737205505             71.456863403                    LEU  52                           h 0.1000                                  147HB2 57.488136292      37.658962250             70.374801636                    LEU  52                           h 0.1000                                  148CG  57.411731720      39.487998962             71.552589417                    LEU  52                           c1                             -0.1000                                  149HG  56.652648926      40.244640350             71.834617615                    LEU  52                           h 0.1000                                  150CD1 58.010108948      38.936943054             72.859535217                    LEU  52                           c3                             -0.3000                                  151HD11    58.475826263      39.735752106             73.467353821                    LEU  52                           h 0.1000                                  152HD12    57.236072540      38.469894409             73.497505188                    LEU  52                           h 0.1000                                  153HD13    58.787303925      38.171112061             72.675750732                    LEU  52                           h 0.1000                                  154CD2 58.517623901      40.187110901             70.742630005                    LEU  52                           c3                             -0.3000                                  155HD21    58.993679047      41.001243591             71.321037292                    LEU  52                           h 0.1000                                  156HD22    59.321178436      39.487018585             70.445312500                    LEU  52                           h 0.1000                                  157HD23    58.125030518      40.647811890             69.818283081                    LEU  52                           h 0.1000                                  158N   54.475013733      36.643035889             69.315246582                    ALA  53                           n -0.5000                                  159CA  53.896503448      35.451416016             68.639259338                    ALA  53                           ca                             0.1200                                  160HN  54.100524902      37.007503510             70.205230713                    ALA  53                           hn                             0.2800                                  161HA  53.553531647      35.773445129             67.634338379                    ALA  53                           h 0.1000                                  162C   52.602260590      34.908508301             69.353744507                    ALA  53                           c&#39;                             0.3800                                  163O   51.589202881      34.828308105             68.650034414                    ALA  53                           o&#39;                             -0.3800                                  164CB  54.970031738      34.364234924             68.394615173                    ALA  53                           c3                             -0.3000                                  165HB1 54.534633636      33.449993134             67.949813843                    ALA  53                           h 0.1000                                  166HB2 55.742454529      34.720954895             67.688003540                    ALA  53                           h 0.1000                                  167HB3 55.500236511      34.068145752             69.316299438                    ALA  53                           h 0.1000                                  168N   52.528438568      34.519321442             70.670852661                    PRO  54                           n -0.4200                                  169CA  51.288402557      33.944232941             71.268875122                    PRO  54                           ca                             0.0600                                  170HA  50.860397339      33.194515228             70.573081970                    PRO  54                           h 0.1000                                  171CD  53.678298950      34.513732910             71.601814270                    PRO  54                           c2                             0.0600                                  172HD1 54.146690369      35.509193420             71.717323303                    PRO  54                           h 0.1000                                  173HD2 54.456859589      33.804187775             71.264945984                    PRO  54                           h 0.1000                                  174C   50.163700104      34.973747253             71.631500244                    PRO  54                           c&#39;                             0.3800                                  175O   50.381851196      36.186935425             71.709472656                    PRO  54                           o&#39;                             -0.3800                                  176CB  51.868888855      33.209735870             72.497810364                    PRO  54                           c2                             -0.2000                                  177HB1 51.140216827      33.071922302             73.319641113                    PRO  54                           h 0.1000                                  178HB2 52.201725006      32.193626404             72.207275391                    PRO  54                           h 0.1000                                  179CG  53.074722290      34.047100067             72.925216675                    PRO  54                           c2                             -0.2000                                  180HG1 52.742275238      34.920612335             73.520271301                    PRO  54                           h 0.1000                                  181HG2 53.794158936      33.482940674             73.547462463                    PRO  54                           h 0.1000                                  182N   48.950717926      34.451553345             71.882225037                    GLY  55                           n -0.5000                                  183CA  47.799011230      35.264900208             72.354187012                    GLY  55                           cg                             0.0200                                  184HN  48.913242340      33.427757263             71.833122253                    GLY  55                           hn                             0.2800                                  185HA1 47.829734802      36.291049957             71.943481445                    GLY  55                           h 0.1000                                  186HA2 46.873668671      34.838825226             71.925086975                    GLY  55                           h 0.1000                                  187C   47.624210358      35.262092590             73.898422241                    GLY  55                           c&#39;                             0.3800                                  188O   47.184692383      34.228916168             74.411125183                    GLY  55                           o&#39;                             -0.3800                                  189N   47.910079956      36.345748901             74.679351807                    PRO  56                           n -0.4200                                  190CA  47.789894104      36.319248199             76.162750244                    PRO  56                           ca                             0.0600                                  191HA  48.177116394      35.361667633             76.567420959                    PRO  56                           h 0.1000                                  192CD  48.602729797      37.547939301             74.184982300                    PRO  56                           c2                             0.0600                                  193HD1 48.046642303      38.065422058             73.380996704                    PRO  56                           h 0.1000                                  194HD2 49.595470428      37.261718750             73.794586182                    PRO  56                           h 0.1000                                  195C   46.326736450      36.529109955             76.663719177                    PRO  56                           c&#39;                             0.3800                                  196O   45.857757568      37.657508850             76.842826843                    PRO  56                           o&#39;                             -0.3800                                  197CB  48.777050018      37.430667877             76.578651428                    PRO  56                           c2                             -0.2000                                  198HB1 48.524734497      37.898471832             77.549873352                    PRO  56                           h 0.1000                                  199HB2 49.795513153      37.009433746             76.691307068                    PRO  56                           h 0.1000                                  200CG  48.752750397      38.431644440             75.422836304                    PRO  56                           c2                             -0.2000                                  201HG1 47.879917145      39.105598450             75.522354126                    PRO  56                           h 0.1000                                  202HG2 49.655212402      39.069591522             75.391311646                    PRO  56                           h 0.1000                                  203N   45.616943359      35.415058136             76.897827148                    HIS  57                           n -0.5000                                  204HN  46.014194489      34.579135895             76.450401306                    HIS  57                           hn                             0.2800                                  205CA  44.212440491      35.434158325             77.393867493                    HIS  57                           ca                             0.1200                                  206HA  43.635601044      36.135776520             76.762809753                    HIS  57                           h 0.1000                                  207C   44.146274567      35.882919312             78.904235840                    HIS  57                           c&#39;                             0.3800                                  208O   44.718753815      35.174930573             79.742973328                    HIS  57                           o&#39;                             -0.3800                                  209CB  43.577262878      34.025093079             77.198188782                    HIS  57                           c2                             -0.2000                                  210HB1 44.242053986      33.250709534             77.626487732                    HIS  57                           h 0.1000                                  211HB2 42.665222168      33.964206696             77.822631836                    HIS  57                           h 0.1000                                  212CG  43.190551718      33.606594086             75.770996094                    HIS  57                           c5                             0.1000                                  213ND1 44.009815216      33.724720001             74.654800415                    HIS  57                           np                             -0.4200                                  214CE1 43.220783234      33.103317261             73.724327087                    HIS  57                           c5                             0.2700                                  215NE2 42.000507355      32.603866577             74.087806702                    HIS  57                           np                             -0.5000                                  216CD2 42.008693695      32.921348572             75.433784485                    HIS  57                           c5                             0.0100                                  217HE1 43.579235077      32.995296478             72.708923340                    HIS  57                           h 0.1300                                  218HE2 41.324546814      32.061363220             73.538429260                    HIS  57                           hn                             0.2800                                  219HD2 41.238662720      32.638732910             76.138023376                    HIS  57                           h 0.1300                                  220N   43.493415833      37.014366150             79.314620972                    PRO  58                           n -0.4200                                  221CA  43.576023102      37.521991730             80.713310242                    PRO  58                           ca                             0.0600                                  222HA  44.640773773      37.575752258             81.019523621                    PRO  58                           h 0.1000                                  223CD  42.828823090      37.960189819             78.395561218                    PRO  58                           c2                             0.0600                                  224HD1 42.088195801      37.471439362             77.735382080                    PRO  58                           h 0.1000                                  225HD2 43.573791504      38.467478070             77.749885559                    PRO  58                           h 0.1000                                  226C   42.782455444      36.660888672             81.747703552                    PRO  58                           c&#39;                             0.3800                                  227O   41.546211243      36.662284851             81.766304016                    PRO  58                           o&#39;                             -0.3800                                  228CB  43.067096710      38.972381592             80.563407898                    PRO  58                           c2                             -0.2000                                  229HB1 42.553604126      39.355644226             81.465919495                    PRO  58                           h 0.1000                                  230HB2 43.922630310      39.652961731             80.382720947                    PRO  58                           h 0.1000                                  231CG  42.156223297      38.958541870             79.333923340                    PRO  58                           c2                             -0.2000                                  232HG1 41.146640778      38.599040985             79.611282349                    PRO  58                           h 0.1000                                  233HG2 42.036239624      39.956802368             78.871780396                    PRO  58                           h 0.1000                                  234N   43.511478424      35.934528351             82.617271423                    ALA  59                           n -0.5000                                  235HN  44.509765625      35.905010223             82.386589050                    ALA  59                           hn                             0.2800                                  236CA  42.913761139      34.926112030             83.544029236                    ALA  59                           ca                             0.1200                                  237HA  42.154701233      34.341350555             82.987930298                    ALA  59                           h 0.1000                                  238C   42.181308746      35.481468201             84.821502686                    ALA  59                           c&#39;                             0.3800                                  239O   42.459964752      35.112228394             85.965652466                    ALA  59                           o&#39;                             -0.3800                                  240CB  44.063701630      33.948829651             83.858291626                    ALA  59                           c3                             -0.3000                                  241HB1 43.708641052      33.104522705             84.478195190                    ALA  59                           h 0.1000                                  242HB2 44.502014160      33.503505707             82.944122314                    ALA  59                           h 0.1000                                  243HB3 44.882900238      34.433902740             84.422813416                    ALA  59                           h 0.1000                                  244N   41.178901672      36.333106995             84.577079773                    ALA  60                           n -0.5000                                  245HN  41.112052917      36.562049866             83.573013306                    ALA  60                           hn                             0.2800                                  246CA  40.189502716      36.803413391             85.578857422                    ALA  60                           ca                             0.1200                                  247HA  40.008514404      35.981742859             86.302818298                    ALA  60                           h 0.1000                                  248C   38.811019897      37.037807465             84.860046387                    ALA  60                           c&#39;                             0.3800                                  249O   37.881835938      36.301105499             85.205276489                    ALA  60                           o&#39;                             -0.3800                                  250CB  40.746566772      37.985549927             86.401313782                    ALA  60                           c3                             -0.3000                                  251HB1 39.997776031      38.372333527             87.116531372                    ALA  60                           h 0.1000                                  252HB2 41.624504089      37.670467377             86.996170044                    ALA  60                           h 0.1000                                  253HB3 41.079444885      38.830841064             85.774902344                    ALA  60                           h 0.1000                                  254N   38.609931946      37.922218323             83.826889038                    PRO  61                           n -0.4200                                  255CA  37.398490906      37.873073578             82.950836182                    PRO  61                           ca                             0.0600                                  256HA  36.485267639      37.839759827             83.576164246                    PRO  61                           h 0.1000                                  257CD  39.552070618      39.001556396             83.460945129                    PRO  61                           c2                             0.0600                                  258HD1 40.590957642      38.653537750             83.311111450                    PRO  61                           h 0.1000                                  259HD2 39.564563751      39.780746460             84.247795105                    PRO  61                           h 0.1000                                  260C   37.256782532      36.700027466             81.909835815                    PRO  61                           c&#39;                             0.3800                                  261O   36.243316650      36.657413483             81.209106445                    PRO  61                           o&#39;                             -0.3800                                  262CB  37.477272034      39.256717682             82.271400452                    PRO  61                           c2                             -0.2000                                  263HB1 36.964195251      39.298637390             81.290786743                    PRO  61                           h 0.1000                                  264HB2 36.981357574      40.016944885             82.906776428                    PRO  61                           h 0.1000                                  265CG  38.972518921      39.562160492             82.163795471                    PRO  61                           c2                             -0.2000                                  266HG1 39.409160614      39.034259796             81.293624878                    PRO  61                           h 0.1000                                  267HG2 39.183399200      40.640045166             82.032295227                    PRO  61                           h 0.1000                                  268N   38.213741302      35.753643036             81.804443359                    SER  62                           n -0.5000                                  269CA  38.144962311      34.600208282             80.863403320                    SER  62                           ca                             0.1200                                  270HN  39.009967804      35.895751953             82.434890747                    SER  62                           hn                             0.2800                                  271HA  37.892318726      34.983673096             79.856529236                    SER  62                           h 0.1000                                  272C   37.085021973      33.524326324             81.273231506                    SER  62                           c&#39;                             0.3800                                  273O   37.382484436      32.625560760             82.070343018                    SER  62                           o&#39;                             -0.3800                                  274CB  39.569152832      33.994293213             80.760513306                    SER  62                           c2                             -0.1700                                  275HB1 39.601100922      33.242229462             79.949050903                    SER  62                           h 0.1000                                  276HB2 40.317050934      34.759819031             80.475799561                    SER  62                           h 0.1000                                  277OG  39.958038330      33.367904663             81.986091614                    SER  62                           oh                             -0.3800                                  278HG  39.157264709      32.928077698             82.316406250                    SER  62                           ho                             0.3500                                  279N   35.853912354      33.643447876             80.748901367                    SER  63                           n -0.5000                                  280CA  34.681579590      32.893772125             81.280097961                    SER  63                           ca                             0.1200                                  281HN  35.734226227      34.507610321             80.200576782                    SER  63                           hn                             0.2800                                  282HA  34.978878021      32.281963348             82.158424377                    SER  63                           h 0.1000                                  283C   34.028987885      31.963005066             80.214485168                    SER  63                           c&#39;                             0.3800                                  284O   33.624385834      32.404701233             79.134857178                    SER  63                           o&#39;                             -0.3800                                  285CB  33.674541473      33.937458038             81.815757751                    SER  63                           c2                             -0.1700                                  286HB1 34.172107697      34.614356995             82.538948059                    SER  63                           h 0.1000                                  287HB2 33.303077698      34.594402313             81.003784180                    SER  63                           h 0.1000                                  288OG  32.576236725      33.301517487             82.471984863                    SER  63                           oh                             -0.3800                                  289HG  32.084590912      32.806625366             81.807708740                    SER  63                           ho                             0.3500                                  290N   33.852622986      30.677625656             80.556045532                    TRP  64                           n -0.5000                                  291CA  33.070865631      29.709415436             79.732810974                    TRP  64                           ca                             0.1200                                  292HN  34.153343201      30.435497284             81.506057739                    TRP  64                           hn                             0.2800                                  293HA  33.375720978      29.840501785             78.676765442                    TRP  64                           h 0.1000                                  294C   31.526346207      29.958730698             79.816452026                    TRP  64                           c&#39;                             0.3800                                  295O   30.936735153      29.911306381             80.900970459                    TRP  64                           o&#39;                             -0.3800                                  296CB  33.498836517      28.253648758             80.086425781                    TRP  64                           c2                             -0.2000                                  297HB1 32.936897276      27.550512314             79.442245483                    TRP  64                           h 0.1000                                  298HB2 34.548812866      25.110004425             79.767990112                    TRP  64                           h 0.1000                                  299CG  33.372638702      27.788063049             81.551361084                    TRP  64                           c5                             0.0000                                  300CD1 32.236022949      27.198472977             82.145393372                    TRP  64                           c5                             0.0100                                  301NE1 32.442039490      26.926628113             83.513259888                    TRP  64                           np                             -0.5000                                  302CE2 33.734771729      27.365074158             83.750877380                    TRP  64                           c5                             0.1100                                  303CD2 34.313194275      27.885219574             82.565856934                    TRP  64                           c5                             0.0000                                  304HD1 31.308589935      27.011001587             81.622375488                    TRP  64                           h 0.1000                                  305HE1 31.781488419      26.532098770             84.192108154                    TRP  64                           hn                             0.2800                                  306CE3 35.633460999      28.410655975             82.581642151                    TRP  64                           cp                             -0.1000                                  307HE3 36.086208344      28.812973022             81.686943054                    TRP  64                           h 0.1000                                  308CZ3 36.338203430      28.401332855             83.786201477                    TRP  64                           cp                             -0.1000                                  309HZ3 37.342418671      28.800062180             83.816780090                    TRP  64                           h 0.1000                                  310CH2 35.766292572      27.887487411             84.956939697                    TRP  64                           cp                             -0.1000                                  311HH2 36.336753845      27.895225525             85.875114441                    TRP  64                           h 0.1000                                  312CZ2 34.469055176      27.367534637             84.959693909                    TRP  64                           cp                             -0.1000                                  313HZ2 34.033626556      26.978828430             85.868453979                    TRP  64                           h 0.1000                                  314N   30.884126663      30.253189087             78.672058105                    GLY  65                           n -0.5000                                  315CA  29.433704376      30.582933426             78.625785828                    GLY  65                           cg                             0.0200                                  316HN  31.490442276      30.324731827             77.837860107                    GLY  65                           hn                             0.2800                                  317HA1 29.049486160      30.927183151             79.604759216                    GLY  65                           h 0.1000                                  318HA2 29.301883698      31.462034225             77.967575073                    GLY  65                           h 0.1000                                  319C   28.566276550      29.436250687             78.049354553                    GLY  65                           c&#39;                             0.3800                                  320O   28.476503372      29.383417130             76.820671082                    GLY  65                           o&#39;                             -0.3800                                  321N   27.919076920      28.520057678             78.830680847                    PRO  66                           n -0.4200                                  322CA  27.254581451      27.307062149             78.266265869                    PRO  66                           ca                             0.0600                                  323HA  28.007204056      26.749544144             77.674038860                    PRO  66                           h 0.1000                                  324CD  27.931732178      28.547025681             80.307373047                    PRO  66                           c2                             0.0600                                  325HD1 27.674114227      29.536535263             80.727989197                    PRO  66                           h 0.1000                                  326HD2 28.930458069      28.264209747             80.693565369                    PRO  66                           h 0.1000                                  327C   25.991449356      27.540506363             77.367637634                    PRO  66                           c&#39;                             0.3800                                  328O   25.234470367      28.498056412             77.550445557                    PRO  66                           o&#39;                             -0.3800                                  329CB  26.947065353      26.487516403             79.540168762                    PRO  66                           c2                             -0.2000                                  330HB1 26.021696091      25.883453349             79.467781067                    PRO  66                           h 0.1000                                  331HB2 27.765922546      25.768106461             79.730659485                    PRO  66                           h 0.1000                                  332CG  26.879997253      27.505201340             80.680580139                    PRO  66                           c2                             -0.2000                                  333HG1 25.878761292      27.974979401             80.732356567                    PRO  66                           h 0.1000                                  334HG2 27.057765961      27.053478241             81.674545288                    PRO  66                           h 0.1000                                  335N   25.764133453      26.624628067             76.406608582                    CYS  67                           n -0.5000                                  336CA  24.578670502      26.665664673             75.512832642                    CYS  67                           ca                             0.1200                                  337HN  26.474828720      25.893449783             76.320098877                    CYS  67                           hn                             0.2800                                  338HA  24.437746048      27.701400757             75.152099609                    CYS  67                           h 0.1000                                  339C   23.256376266      26.130277634             76.174392700                    CYS  67                           c&#39;                             0.3800                                  340O   23.219629288      24.940492630             76.516906738                    CYS  67                           o&#39;                             -0.3800                                  341CB  24.900175095      25.815908432             74.260444641                    CYS  67                           c2                             -0.3000                                  342HB1 25.807971954      26.178848267             73.749794006                    CYS  67                           h 0.1000                                  343HB2 25.105169296      24.761932373             74.532623291                    CYS  67                           h 0.1000                                  344SG  23.472158432      25.844451904             73.133270264                    CYS  67                           s1                             0.1000                                  345N   22.124137878      26.895683289             76.264381409                    PRO  68                           n -0.4200                                  346CA  20.786550522      26.297697067             76.529830933                    PRO  68                           ca                             0.0600                                  347HA  20.877141953      25.506265640             77.300582886                    PRO  68                           h 0.1000                                  348CD  22.161409378      28.364057541             76.432929993                    PRO  68                           c2                             0.0600                                  349HD1 22.628620148      28.901371002             75.585960388                    PRO  68                           h 0.1000                                  350HD2 22.732339859      28.631174088             77.345329285                    PRO  68                           h 0.1000                                  351C   20.190311432      25.593938828             75.255737305                    PRO  68                           c&#39;                             0.3800                                  352O   20.566764832      24.451507568             74.984413147                    PRO  68                           o&#39;                             -0.3800                                  353CB  20.033475876      27.483673096             77.173645020                    PRO  68                           c2                             -0.2000                                  354HB1 18.940057755      27.449979782             77.017181396                    PRO  68                           h 0.1000                                  355HB2 20.183664322      27.458950043             78.271354675                    PRO  68                           h 0.1000                                  356CG  20.687761307      28.746078411             76.604209900                    PRO  68                           c2                             -0.2000                                  357HG1 20.234010696      29.017192841             75.632743835                    PRO  68                           h 0.1000                                  358HG2 20.559530258      29.023554230             77.265640259                    PRO  68                           h 0.1000                                  359N   19.297069550      26.226366043             74.460205078                    ARG+ 69                           n -0.5000                                  360CA  18.727945328      25.594141006             73.229873657                    ARG+ 69                           cd                             0.1200                                  361HN  18.899488449      27.074655533             74.874603271                    ARG+ 69                           hn                             0.2800                                  362HA  19.468439102      24.889890671             72.798027039                    ARG+ 69                           h 0.1000                                  363C   18.426181793      26.666866302             72.127845764                    ARG+ 69                           c&#39;                             0.3800                                  364O   17.302417755      27.170328140             72.057907104                    ARG+ 69                           o&#39;                             -0.3800                                  365CB  17.487716675      24.741283417             73.645401001                    ARG+ 69                           c2                             -0.2000                                  366HB1 17.790594101      24.038330078             74.447227478                    ARG+ 69                           h 0.1100                                  367HB2 16.742151260      25.409061432             74.119949341                    ARG+ 69                           h 0.1100                                  368CG  16.806570053      23.930654526             72.510940552                    ARG+ 69                           c2                             -0.2000                                  369HG1 16.500089645      24.624885559             71.702163696                    ARG+ 69                           h 0.1300                                  370HG2 17.539186478      23.235509872             72.053100586                    ARG+ 69                           h 0.1300                                  371CD  15.574314117      23.148860931             73.007453918                    ARG+ 69                           c2                             -0.0900                                  372HD1 15.890284538      22.374292374             73.738624573                    ARG+ 69                           h 0.1300                                  373HD2 14.902976036      23.843069077             73.554016113                    ARG+ 69                           h 0.1300                                  374NE  14.865127563      22.521680832             71.855873108                    ARG+ 69                           n1                             -0.5000                                  375HE  15.293711662      22.507183075             70.926025391                    ARG+ 69                           hn                             0.3600                                  376CZ  13.645489693      21.980854034             71.902374268                    ARG+ 69                           cr                             0.4500                                  377NH1 13.127552986      21.522832870             70.798370361                    ARG+ 69                           n2                             -0.5000                                  378HH11    13.689088821      21.608518600             69.948852539                    ARG+ 69                           hn                             0.3600                                  379HH12    12.188611031      21.122539520             70.853851318                    ARG+ 69                           hn                             0.3600                                  380NH2 12.936479568      21.886768341             73.000465393                    ARG+ 69                           n2                             -0.5000                                  381HH21    12.008401871      21.462900162             72.952354431                    ARG+ 69                           hn                             0.3600                                  382HH22    13.405644417      22.251142502             73.831558228                    ARG+ 69                           hn                             0.3600                                  383N   19.430337906      26.966600418             71.273384094                    ARG+ 70                           n -0.5000                                  384CA  19.316965103      27.784936905             70.016807556                    ARG+ 70                           ca                             0.1200                                  385HN  20.317523956      26.522169576             71.527793884                    ARG+ 70                           hn                             0.2800                                  386HA  19.139877319      27.013025284             69.241798401                    ARG+ 70                           h 0.1000                                  387C   20.690151215      28.397680283             69.573341370                    ARG+ 70                           c&#39;                             0.3800                                  388O   21.267679214      27.963806152             68.579154968                    ARG+ 70                           o&#39;                             -0.3800                                  389CB  18.103752136      28.753881454             69.796676636                    ARG+ 70                           c2                             -0.2000                                  390HB1 18.134477615      29.133897781             68.754875183                    ARG+ 70                           h 0.1100                                  391HB2 17.183977127      28.135446548             69.816154480                    ARG+ 70                           h 0.1100                                  392CG  17.944366455      29.952959061             70.767364502                    ARG+ 70                           c2                             -0.2000                                  393HG1 18.201717377      29.630409241             71.795295715                    ARG+ 70                           h 0.1300                                  394HG2 18.686578751      30.737569809             70.523498535                    ARG+ 70                           h 0.1300                                  395CD  16.516407013      30.528032303             70.757499695                    ARG+ 70                           c2                             -0.0900                                  396HD1 16.205293655      30.812314987             69.732498169                    ARG+ 70                           h 0.1300                                  397HD2 15.804359436      29.724859238             71.041206360                    ARG+ 70                           h 0.1300                                  398NE  16.396289825      31.632083893             71.751823425                    ARG+ 70                           n1                             -0.5000                                  399HE  16.382929398      31.418577194             72.754409790                    ARG+ 70                           hn                             0.3600                                  400CZ  16.270032883      32.931114197             71.475357056                    ARG+ 70                           cr                             0.4500                                  401NH1 16.119342804      33.758121490             72.470024109                    ARG+ 70                           n2                             -0.5000                                  402HH11    16.097789764      33.352241516             73.407394409                    ARG+ 70                           hn                             0.3600                                  403HH12    16.020824432      34.751869202             72.242614746                    ARG+ 70                           hn                             0.3600                                  404NH2 16.295974731      33.427280426             70.262794495                    ARG+ 70                           n2                             -0.5000                                  405HH21    16.191591263      34.436088562             70.142570496                    ARG+ 70                           hn                             0.3600                                  406HH22    16.439420972      32.732715607             69.527351379                    ARG+ 70                           hn                             0.3600                                  407N   21.215826035      29.506198883             70.104598999                    TYRC 71                           n -0.5000                                  408HN  21.692993164      29.961484909             69.319816589                    TYRC 71                           hn                             0.2800                                  409CA  22.037544250      29.469150543             71.348724365                    TYRC 71                           ca                             0.1200                                  410HA  22.727062225      28.601654053             71.296867371                    TYRC 71                           h 0.1000                                  411C   21.230804443      29.295030594             72.663772583                    TYRC 71                           c&#39;                             0.4100                                  412OXT 20.420524597      30.105148315             73.113685608                    TYRC 71                           o&#39;                             -0.3800                                  413O   21.522335052      28.107995987             73.273979187                    TYRC 71                           oh                             -0.3800                                  414HO  20.994127274      28.000021444             74.066841125                    TYRC 71                           ho                             0.3500                                  415CB  22.938613892      30.740638733             71.402084351                    TYRC 71                           c2                             -0.2000                                  416HB1 22.321226120      31.652799606             71.283157349                    TYRC 71                           h 0.1000                                  417HB2 23.363500595      30.853305817             72.420455933                    TYRC 71                           h 0.1000                                  418CG  24.110603333      30.760416031             70.402580261                    TYRC 71                           cp                             0.0000                                  419CD1 23.933057785      31.274461746             69.111869812                    TYRC 71                           cp                             -0.1000                                  420HD1 22.977622986      31.679159164             68.809974670                    TYRC 71                           h 0.1000                                  421CE1 24.985538483      31.264329910             68.201301575                    TYRC 71                           cp                             -0.1000                                  422HE1 24.833002090      31.650396347             67.203536987                    TYRC 71                           h 0.1000                                  423CZ  26.227394104      30.757091522             68.577186584                    TYRC 71                           cp                             0.0300                                  424OH  27.265848160      30.762763977             67.686424255                    TYRC 71                           oh                             -0.3800                                  425HH  26.966634750      31.154380798             66.863937378                    TYRC 71                           ho                             0.3500                                  426CE2 26.415199280      30.251981735             69.859985352                    TYRC 71                           cp                             -0.1000                                  427HE2 27.377700806      29.852491379             70.147377014                    TYRC 71                           h 0.1000                                  428CD2 25.360828400      30.253871918             70.770927429                    TYRC 71                           cp                             -0.1000                                  429HD2 25.521846771      29.846044540             71.760574341                    TYRC 71                           h 0.1000                                  430__________________________________________________________________________ 
    
     
         __________________________________________________________________________#             SEQUENCE LISTING- (1) GENERAL INFORMATION:-    (iii) NUMBER OF SEQUENCES: 8- (2) INFORMATION FOR SEQ ID NO:1:-      (i) SEQUENCE CHARACTERISTICS:#acids    (A) LENGTH: 476 amino     (B) TYPE: amino acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: protein-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:- Leu Gln Pro Gly Ala Glu Val Pro Val Val Tr - #p Ala Gln Glu Gly Ala#                15- Pro Ala Gln Leu Pro Cys Ser Pro Thr Ile Pr - #o Leu Gln Asp Leu Ser#            30- Leu Leu Arg Arg Ala Gly Val Thr Trp Gln Hi - #s Gln Pro Asp Ser Gly#        45- Pro Pro Ala Ala Ala Pro Gly His Pro Leu Al - #a Pro Gly Pro His Pro#    60- Ala Ala Pro Ser Ser Trp Gly Pro Arg Pro Ar - #g Arg Tyr Thr Val Leu#80- Ser Val Gly Pro Gly Gly Leu Arg Ser Gly Ar - #g Leu Pro Leu Gln Pro#                95- Arg Val Gln Leu Asp Glu Arg Gly Arg Gln Ar - #g Gly Asp Phe Ser Leu#           110- Trp Leu Arg Pro Ala Arg Arg Ala Asp Ala Gl - #y Glu Tyr Arg Ala Ala#       125- Val His Leu Arg Asp Arg Ala Leu Ser Cys Ar - #g Leu Arg Leu Arg Leu#   140- Gly Gln Ala Ser Met Thr Ala Ser Pro Pro Gl - #y Ser Leu Arg Ala Ser145                 1 - #50                 1 - #55                 1 -#60- Asp Trp Val Ile Leu Asn Cys Ser Phe Ser Ar - #g Pro Asp Arg Pro Ala#               175- Ser Val His Trp Phe Arg Asn Arg Gly Gln Gl - #y Arg Val Pro Val Arg#           190- Glu Ser Pro His His His Leu Ala Glu Ser Ph - #e Leu Phe Leu Pro Gln#       205- Val Ser Pro Met Asp Ser Gly Pro Trp Gly Cy - #s Ile Leu Thr Tyr Arg#   220- Asp Gly Phe Asn Val Ser Ile Met Tyr Asn Le - #u Thr Val Leu Gly Leu225                 2 - #30                 2 - #35                 2 -#40- Glu Pro Pro Thr Pro Leu Thr Val Tyr Ala Gl - #y Ala Gly Ser Arg Val#               255- Gly Leu Pro Cys Arg Leu Pro Ala Gly Val Gl - #y Thr Arg Ser Phe Leu#           270- Thr Ala Lys Trp Thr Pro Pro Gly Gly Gly Pr - #o Asp Leu Leu Val Thr#       285- Gly Asp Asn Gly Asp Phe Thr Leu Arg Leu Gl - #u Asp Val Ser Gln Ala#   300- Gln Ala Gly Thr Tyr Thr Cys His Ile His Le - #u Gln Glu Gln Gln Leu305                 3 - #10                 3 - #15                 3 -#20- Asn Ala Thr Val Thr Leu Ala Ile Ile Thr Va - #l Thr Pro Lys Ser Phe#               335- Gly Ser Pro Gly Ser Leu Gly Lys Leu Leu Cy - #s Glu Val Thr Pro Val#           350- Ser Gly Gln Glu Arg Phe Val Trp Ser Ser Le - #u Asp Thr Pro Ser Gln#       365- Arg Ser Phe Ser Gly Pro Trp Leu Glu Ala Gl - #n Glu Ala Gln Leu Leu#   380- Ser Gln Pro Trp Gln Cys Gln Leu Tyr Gln Gl - #y Glu Arg Leu Leu Gly385                 3 - #90                 3 - #95                 4 -#00- Ala Ala Val Tyr Phe Thr Glu Leu Ser Ser Pr - #o Gly Ala Gln Arg Ser#               415- Gly Arg Ala Pro Gly Ala Leu Pro Ala Gly Hi - #s Leu Leu Leu Phe Leu#           430- Thr Leu Gly Val Leu Ser Leu Leu Leu Leu Va - #l Thr Gly Ala Phe Gly#       445- Phe His Leu Trp Arg Arg Gln Trp Arg Pro Ar - #g Arg Phe Ser Ala Leu#   460- Glu Gln Gly Ile His Pro Arg Arg Leu Arg Al - #a Arg465                 4 - #70                 4 - #75- (2) INFORMATION FOR SEQ ID NO:2:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 32 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:#          32      ACCA TAGGAGAGAT GT- (2) INFORMATION FOR SEQ ID NO:3:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 40 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:#    40            ACCC AGAACAGTGA GGTTATACAT- (2) INFORMATION FOR SEQ ID NO:4:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 34 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:#        34        GCTA GACAGCTCTG TGAA- (2) INFORMATION FOR SEQ ID NO:5:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 37 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:#      37          CCAG ACCATAGGAG AGATGTG- (2) INFORMATION FOR SEQ ID NO:6:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 37 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:#      37          CGAG GCCTGGCCCA GGCGCAG- (2) INFORMATION FOR SEQ ID NO:7:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 35 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:#       35         CAGA ACAGTGAGGT TATAC- (2) INFORMATION FOR SEQ ID NO:8:-      (i) SEQUENCE CHARACTERISTICS:#pairs    (A) LENGTH: 35 base     (B) TYPE: nucleic acid     (C) STRANDEDNESS: single     (D) TOPOLOGY: linear-     (ii) MOLECULE TYPE: cDNA-     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:#       35         TGGG CTAGACAGCT CTGTG__________________________________________________________________________