data_5743 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Backbone 1H, 13C and 15N Chemical Shift Assignments for the extracellular domain of b-dystroglycan (654-750) ; _BMRB_accession_number 5743 _BMRB_flat_file_name bmr5743.str _Entry_type original _Submission_date 2003-03-17 _Accession_date 2003-03-17 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details . loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bozzi Manuela . . 2 Brancaccio Andrea . . 3 Paci Maurizio . . 4 Cicero Daniel O. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 124 "13C chemical shifts" 142 "15N chemical shifts" 76 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2003-12-18 original author . stop_ _Original_release_date 2003-12-18 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Structural Characterization by NMR of the Natively Unfolded Extracellular Domain of beta-Dystroglycan: Toward the Identification of the Binding Epitope for alpha-Dystroglycan ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 14622018 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bozzi Manuela . . 2 Bianchi Marzia . . 3 Sciandra Francesca . . 4 Paci Maurizio . . 5 Giardina Bruno . . 6 Brancaccio Andrea . . 7 Cicero Daniel O. . stop_ _Journal_abbreviation Biochemistry _Journal_volume 42 _Journal_issue 46 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 13717 _Page_last 13724 _Year 2003 _Details . save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Winder S.J. Trends Biochem Sci. 2001 Feb;26(2):118-24 ; _Citation_title 'The complexities of dystroglycan.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11166570 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Winder 'S. J.' J. . stop_ _Journal_abbreviation 'Trends Biochem. Sci.' _Journal_name_full 'Trends in biochemical sciences' _Journal_volume 26 _Journal_issue 2 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 118 _Page_last 124 _Year 2001 _Details ; The notion of dystroglycan as a simple laminin-binding receptor is increasingly being challenged. New roles and new binding partners are continually emerging. Recent structural advances have provided exciting new insights into the precise molecular interactions between dystroglycan and other key components of the dystroglycan complex. Coupled with an increasing understanding of dystroglycan function at the molecular level, we are finally beginning to probe the complexities of dystroglycan, not only in disease, but in development, adhesion and signalling. ; save_ save_ref_2 _Saveframe_category citation _Citation_full ; Boffi A, Bozzi M, Sciandra F, Woellner C, Bigotti MG, Ilari A, Brancaccio A. Biochim Biophys Acta. 2001 Mar 9;1546(1):114-21 ; _Citation_title 'Plasticity of secondary structure in the N-terminal region of beta-dystroglycan.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11257514 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Boffi A. . . 2 Bozzi M. . . 3 Sciandra F. . . 4 Woellner C. . . 5 Bigotti 'M. G.' G. . 6 Ilari A. . . 7 Brancaccio A. . . stop_ _Journal_abbreviation 'Biochim. Biophys. Acta' _Journal_name_full 'Biochimica et biophysica acta' _Journal_volume 1546 _Journal_issue 1 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 114 _Page_last 121 _Year 2001 _Details ; The secondary structure content of the N-terminal extracellular domain of beta-dystroglycan (a recombinant fragment extending from positions 654 to 750) has been quantitatively determined by means of CD and FTIR spectroscopies. The elements of secondary structure, namely an 8-10 residue long alpha-helix (10%) and two beta-strands (24%) have been assigned to specific amino acid sequences by means of a GOR constrained prediction method. The remaining 66% of the whole sequence is classified as turns or unordered. The temperature dependence of CD and FTIR spectra has been investigated in detail. A reversible, non-cooperative thermal transition is observed with both CD and FTIR spectroscopies up to 95 degrees C. The profile of the transition is typical of the unfolding of isolated peptides and corresponds to the progressive loss of the secondary structure elements of the protein with no evidence for collapsing phenomena, typical of globular proteins, upon heating. ; save_ save_ref_3 _Saveframe_category citation _Citation_full ; Sciandra F, Schneider M, Giardina B, Baumgartner S, Petrucci TC, Brancaccio A. Eur J Biochem 2001 Aug;268(16):4590-7 ; _Citation_title 'Identification of the beta-dystroglycan binding epitope within the C-terminal region of alpha-dystroglycan.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11502221 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Sciandra F. . . 2 Schneider M. . . 3 Giardina B. . . 4 Baumgartner S. . . 5 Petrucci 'T. C.' C. . 6 Brancaccio A. . . stop_ _Journal_abbreviation 'Eur. J. Biochem.' _Journal_name_full 'European journal of biochemistry / FEBS' _Journal_volume 268 _Journal_issue 16 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 4590 _Page_last 4597 _Year 2001 _Details ; Dystroglycan is a receptor for extracellular matrix proteins that plays a crucial role during embryogenesis in addition to adult tissue stabilization. A precursor product of a single gene is post-translationally cleaved to form two different subunits, alpha and beta. The extracellular alpha-dystroglycan is a membrane-associated, highly glycosylated protein that binds to various extracellular matrix molecules, whereas the transmembrane beta-dystroglycan binds, via its cytosolic domain, to dystrophin and many other proteins. alpha- and beta-Dystroglycan interact tightly but noncovalently. We have previously shown that the N-terminal region of beta-dystroglycan, beta-DG(654-750), binds to the C-terminal region of murine alpha-dystroglycan independently from glycosylation. Preparing a series of deleted recombinant fragments and using solid-phase binding assays, the C-terminal sequence of alpha-dystroglycan containing the binding epitope for beta-dystroglycan has been defined more precisely. We found that a region of 36 amino acids, from position 550-585, is required for binding the extracellular region, amino acids 654-750 of beta-dystroglycan. Recently, a dystroglycan-like gene was identified in Drosophila that showed a moderate degree of conservation with vertebrate dystroglycan (31% identity, 48% similarity). Surprisingly, the Drosophila sequence contains a region showing a higher degree of identity and conservation (45% and 66%) that coincides with the 550-585 sequence of vertebrate alpha-dystroglycan. We have expressed this Drosophila dystroglycan fragment and measured its binding to the extracellular region of vertebrate (murine) beta-dystroglycan (Kd = 6 +/- 1 microM). These data confirm the proper identification of the beta-dystroglycan binding epitope and stress the importance of this region during evolution. This finding might help the rational design of dystroglycan-specific binding drugs, that could have important biomedical applications. ; save_ save_ref_4 _Saveframe_category citation _Citation_full ; Bozzi M, Veglia G, Paci M, Sciandra F, Giardina B, Brancaccio A. FEBS Lett 2001 Jun 22;499(3):210-4 ; _Citation_title 'A synthetic peptide corresponding to the 550-585 region of alpha-dystroglycan binds beta-dystroglycan as revealed by NMR spectroscopy.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11423118 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Bozzi M. . . 2 Veglia G. . . 3 Paci M. . . 4 Sciandra F. . . 5 Giardina B. . . 6 Brancaccio A. . . stop_ _Journal_abbreviation 'FEBS Lett.' _Journal_name_full 'FEBS letters' _Journal_volume 499 _Journal_issue 3 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 210 _Page_last 214 _Year 2001 _Details ; We have probed the binding of a synthetic peptide corresponding to the region 550-585 of the alpha subunit of dystroglycan with a recombinant protein fragment corresponding to the N-terminal extracellular region of beta-dystroglycan (654-750), using NMR in solution. In a 30:1 molar ratio, the peptide binds to the recombinant protein fragment in the fast/intermediate exchange regime. By monitoring the peptide intra-residue HN-Halpha peak volumes of the 2D TOCSY NMR spectra, both in the absence and in the presence of the recombinant fragment, we determined the differential binding affinities of each amino acid. We found that the residues in the region 550-565 (SWVQFNSNSQLMYGLP) are more influenced by the presence of the protein, whereas the C-terminal portion is marginally involved. These NMR results have been confirmed by solid-phase binding assays. ; save_ save_ref_5 _Saveframe_category citation _Citation_full ; Panchal SC, Bhavesh NS, Hosur RV J Biomol NMR 2001 Jun;20(2):135-47 ; _Citation_title 'Improved 3D triple resonance experiments, HNN and HN(C)N, for HN and 15N sequential correlations in (13C, 15N) labeled proteins: application to unfolded proteins.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11495245 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Panchal 'S. C.' C. . 2 Bhavesh 'N. S.' S. . 3 Hosur 'R. V.' V. . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_name_full 'Journal of biomolecular NMR' _Journal_volume 20 _Journal_issue 2 _Journal_CSD . _Book_title . _Book_chapter_title . _Book_volume . _Book_series . _Book_publisher . _Book_publisher_city . _Book_ISBN . _Conference_title . _Conference_site . _Conference_state_province . _Conference_country . _Conference_start_date . _Conference_end_date . _Conference_abstract_number . _Thesis_institution . _Thesis_institution_city . _Thesis_institution_country . _Page_first 135 _Page_last 147 _Year 2001 _Details ; Two triple resonance experiments, HNN and HN(C)N, are presented which correlate HN and 15N resonances sequentially along the polypeptide chain of a doubly (13C, 15N) labeled protein. These incorporate several improvements over the previously published sequences for a similar purpose and have several novel features. The spectral characteristics enable direct identification of certain triplets of residues, which provide many starting points for the sequential assignment procedure. The experiments are sensitive and their utility has been demonstrated with a 22 kDa protein under unfolding conditions where most of the standard triple resonance experiments such as HNCA, CBCANH etc. have limited success because of poor amide, Calpha and Cbeta chemical shift dispersions. ; save_ ################################## # Molecular system description # ################################## save_system_b-DG(654-750) _Saveframe_category molecular_system _Mol_system_name 'extracellular domain of beta subunit of dystroglycan' _Abbreviation_common b-DG(654-750) _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'extracellular domain of beta subunit of dystroglycan, b-DG(654-750)' $b-DG(654-750) stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all free' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_b-DG(654-750) _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common 'extracellular domain of beta subunit of dystroglycan' _Abbreviation_common b-DG(654-750) _Molecular_mass 10528.72 _Mol_thiol_state 'all free' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 99 _Mol_residue_sequence ; GSSIVVEWTNNTLPLEPCPK EQIIGLSRRIADENGKPRPA FSNALEPDFKALSIAVTGSG SCRHLQFIPVAPPSPGSSAA PATEVPDRDPEKSSEDDVY ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 -2 GLY 2 -1 SER 3 654 SER 4 655 ILE 5 656 VAL 6 657 VAL 7 658 GLU 8 659 TRP 9 660 THR 10 661 ASN 11 662 ASN 12 663 THR 13 664 LEU 14 665 PRO 15 666 LEU 16 667 GLU 17 668 PRO 18 669 CYS 19 670 PRO 20 671 LYS 21 672 GLU 22 673 GLN 23 674 ILE 24 675 ILE 25 676 GLY 26 677 LEU 27 678 SER 28 679 ARG 29 680 ARG 30 681 ILE 31 682 ALA 32 683 ASP 33 684 GLU 34 685 ASN 35 686 GLY 36 687 LYS 37 688 PRO 38 689 ARG 39 690 PRO 40 691 ALA 41 692 PHE 42 693 SER 43 694 ASN 44 695 ALA 45 696 LEU 46 697 GLU 47 698 PRO 48 699 ASP 49 700 PHE 50 701 LYS 51 702 ALA 52 703 LEU 53 704 SER 54 705 ILE 55 706 ALA 56 707 VAL 57 708 THR 58 709 GLY 59 710 SER 60 711 GLY 61 712 SER 62 713 CYS 63 714 ARG 64 715 HIS 65 716 LEU 66 717 GLN 67 718 PHE 68 719 ILE 69 720 PRO 70 721 VAL 71 722 ALA 72 723 PRO 73 724 PRO 74 725 SER 75 726 PRO 76 727 GLY 77 728 SER 78 729 SER 79 730 ALA 80 731 ALA 81 732 PRO 82 733 ALA 83 734 THR 84 735 GLU 85 736 VAL 86 737 PRO 87 738 ASP 88 739 ARG 89 740 ASP 90 741 PRO 91 742 GLU 92 743 LYS 93 744 SER 94 745 SER 95 746 GLU 96 747 ASP 97 748 ASP 98 749 VAL 99 750 TYR stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-01-28 loop_ _Database_name _Database_accession_code _Database_entry_mol_name _Sequence_query_to_submitted_percentage _Sequence_subject_length _Sequence_identity _Sequence_positive _Sequence_homology_expectation_value DBJ BAC29332 "unnamed protein product [Mus musculus]" 97.98 893 100.00 100.00 3.97e-57 DBJ BAC30105 "unnamed protein product [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 DBJ BAC35576 "unnamed protein product [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 GB AAA99779 "dystroglycan 1 [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 GB AAC52853 "dystroglycan, partial [Mus musculus]" 97.98 274 100.00 100.00 1.28e-60 GB AAH07150 "Dag1 protein [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 GB EDL21272 "dystroglycan 1 [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 GB EGW08936 "Dystroglycan [Cricetulus griseus]" 97.98 893 96.91 100.00 7.02e-56 REF NP_001263410 "dystroglycan precursor [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 REF NP_001263411 "dystroglycan precursor [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 REF NP_001263414 "dystroglycan precursor [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 REF NP_001263415 "dystroglycan precursor [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 REF NP_001263421 "dystroglycan precursor [Mus musculus]" 97.98 893 100.00 100.00 3.78e-57 SP Q62165 "RecName: Full=Dystroglycan; AltName: Full=Dystrophin-associated glycoprotein 1; Contains: RecName: Full=Alpha-dystroglycan; Sho" 97.98 893 100.00 100.00 3.78e-57 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Tissue $b-DG(654-750) mouse 10090 Eukaryota Metazoa Mus musculus 'skeletal muscle' stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Vector_name $b-DG(654-750) 'recombinant technology' 'E. coli' Escherichia coli BL21(DE3) . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $b-DG(654-750) . mM 0.3 0.5 '[U-13C; U-15N]' 'sodium phosphate' 20 mM . . . 'sodium chloride' 0.15 M . . . stop_ save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 400 _Details . save_ save_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 700 _Details . save_ ############################# # NMR applied experiments # ############################# save_3D_15N-TOCSY-HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 15N-TOCSY-HSQC' _Sample_label $sample_1 save_ save_3D_HNN_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNN' _Sample_label $sample_1 save_ save_3D_HNCO_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCO' _Sample_label $sample_1 save_ save_3D_HNCA_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCA' _Sample_label $sample_1 save_ save_3D_HN(CO)CA_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HN(CO)CA' _Sample_label $sample_1 save_ save_3D_CBCANH_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D CBCANH' _Sample_label $sample_1 save_ save_3D_CBCA(CO)NH_7 _Saveframe_category NMR_applied_experiment _Experiment_name '3D CBCA(CO)NH' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_Ex_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 6.5 0.2 n/a temperature 298 0.01 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference _Saveframe_category chemical_shift_reference _Details . loop_ _Mol_common_name _Atom_type _Atom_isotope_number _Atom_group _Chem_shift_units _Chem_shift_value _Reference_method _Reference_type _External_reference_sample_geometry _External_reference_location _External_reference_axis _Indirect_shift_ratio DSS C 13 'methyl protons' ppm 0.0 . indirect . . . 0.251449530 DSS H 1 'methyl protons' ppm 0.0 internal direct . . . 1.000000000 DSS N 15 'methyl protons' ppm 0.0 . indirect . . . 0.101329118 stop_ save_ ################################### # Assigned chemical shift lists # ################################### ################################################################### # Chemical Shift Ambiguity Index Value Definitions # # # # The values other than 1 are used for those atoms with different # # chemical shifts that cannot be assigned to stereospecific atoms # # or to specific residues or chains. # # # # Index Value Definition # # # # 1 Unique (including isolated methyl protons, # # geminal atoms, and geminal methyl # # groups with identical chemical shifts) # # (e.g. ILE HD11, HD12, HD13 protons) # # 2 Ambiguity of geminal atoms or geminal methyl # # proton groups (e.g. ASP HB2 and HB3 # # protons, LEU CD1 and CD2 carbons, or # # LEU HD11, HD12, HD13 and HD21, HD22, # # HD23 methyl protons) # # 3 Aromatic atoms on opposite sides of # # symmetrical rings (e.g. TYR HE1 and HE2 # # protons) # # 4 Intraresidue ambiguities (e.g. LYS HG and # # HD protons or TRP HZ2 and HZ3 protons) # # 5 Interresidue ambiguities (LYS 12 vs. LYS 27) # # 6 Intermolecular ambiguities (e.g. ASP 31 CA # # in monomer 1 and ASP 31 CA in monomer 2 # # of an asymmetrical homodimer, duplex # # DNA assignments, or other assignments # # that may apply to atoms in one or more # # molecule in the molecular assembly) # # 9 Ambiguous, specific ambiguity not defined # # # ################################################################### save_shift_set_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $Ex_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'extracellular domain of beta subunit of dystroglycan, b-DG(654-750)' _Text_data_format . _Text_data . loop_ _Atom_shift_assign_ID _Residue_author_seq_code _Residue_seq_code _Residue_label _Atom_name _Atom_type _Chem_shift_value _Chem_shift_value_error _Chem_shift_ambiguity_code 1 . 4 ILE C C 176.20 0.05 1 2 . 4 ILE N N 122.10 0.05 1 3 . 5 VAL H H 8.22 0.03 1 4 . 5 VAL HA H 4.07 0.03 1 5 . 5 VAL C C 176.14 0.05 1 6 . 5 VAL CA C 62.27 0.05 1 7 . 5 VAL N N 125.12 0.05 1 8 . 6 VAL H H 8.24 0.03 1 9 . 6 VAL C C 176.01 0.05 1 10 . 6 VAL CA C 62.17 0.05 1 11 . 6 VAL N N 124.75 0.05 1 12 . 7 GLU H H 8.38 0.03 1 13 . 7 GLU HA H 4.30 0.03 1 14 . 7 GLU C C 176.29 0.05 1 15 . 7 GLU CA C 56.47 0.05 1 16 . 7 GLU N N 124.50 0.05 1 17 . 8 TRP H H 8.26 0.03 1 18 . 8 TRP HA H 4.72 0.03 1 19 . 8 TRP C C 176.54 0.05 1 20 . 8 TRP CA C 57.37 0.05 1 21 . 8 TRP N N 122.44 0.05 1 22 . 9 THR H H 7.91 0.03 1 23 . 9 THR HA H 4.23 0.03 1 24 . 9 THR C C 176.50 0.05 1 25 . 9 THR CA C 61.67 0.05 1 26 . 9 THR N N 115.16 0.05 1 27 . 10 ASN H H 8.24 0.03 1 28 . 10 ASN C C 175.17 0.05 1 29 . 10 ASN CA C 53.57 0.05 1 30 . 10 ASN N N 120.05 0.05 1 31 . 11 ASN H H 8.35 0.03 1 32 . 11 ASN C C 175.49 0.05 1 33 . 11 ASN CA C 53.57 0.05 1 34 . 11 ASN N N 118.86 0.05 1 35 . 12 THR H H 8.06 0.03 1 36 . 12 THR C C 174.51 0.05 1 37 . 12 THR CA C 62.17 0.05 1 38 . 12 THR N N 113.67 0.05 1 39 . 13 LEU H H 8.13 0.03 1 40 . 13 LEU HA H 4.58 0.03 1 41 . 13 LEU CA C 53.37 0.05 1 42 . 13 LEU N N 125.39 0.05 1 43 . 14 PRO C C 176.79 0.05 1 44 . 14 PRO CA C 62.97 0.05 1 45 . 15 LEU H H 8.21 0.03 1 46 . 15 LEU HA H 4.27 0.03 1 47 . 15 LEU C C 177.41 0.05 1 48 . 15 LEU CA C 54.67 0.05 1 49 . 15 LEU N N 121.75 0.05 1 50 . 16 GLU H H 8.32 0.03 1 51 . 16 GLU CA C 54.27 0.05 1 52 . 16 GLU N N 122.39 0.05 1 53 . 19 PRO C C 177.13 0.05 1 54 . 20 LYS H H 8.47 0.03 1 55 . 20 LYS C C 177.09 0.05 1 56 . 20 LYS CA C 56.87 0.05 1 57 . 20 LYS N N 121.37 0.05 1 58 . 21 GLU H H 8.47 0.03 1 59 . 21 GLU CA C 56.87 0.05 1 60 . 21 GLU N N 120.68 0.05 1 61 . 22 GLN C C 174.47 0.05 1 62 . 22 GLN N N 117.89 0.05 1 63 . 23 ILE H H 8.21 0.03 1 64 . 23 ILE C C 176.53 0.05 1 65 . 23 ILE CA C 61.27 0.05 1 66 . 23 ILE N N 122.18 0.05 1 67 . 24 ILE H H 8.27 0.03 1 68 . 24 ILE HA H 4.11 0.03 1 69 . 24 ILE C C 176.95 0.05 1 70 . 24 ILE CA C 61.47 0.05 1 71 . 24 ILE N N 125.10 0.05 1 72 . 25 GLY H H 8.40 0.03 1 73 . 25 GLY HA2 H 3.92 0.03 1 74 . 25 GLY HA3 H 3.92 0.03 1 75 . 25 GLY C C 174.51 0.05 1 76 . 25 GLY CA C 45.57 0.05 1 77 . 25 GLY N N 111.81 0.05 1 78 . 26 LEU H H 8.05 0.03 1 79 . 26 LEU C C 177.66 0.05 1 80 . 26 LEU CA C 55.27 0.05 1 81 . 26 LEU N N 121.33 0.05 1 82 . 27 SER H H 8.34 0.03 1 83 . 27 SER HA H 4.44 0.03 1 84 . 27 SER C C 173.76 0.05 1 85 . 27 SER CA C 58.37 0.05 1 86 . 27 SER N N 116.92 0.05 1 87 . 28 ARG H H 7.99 0.03 1 88 . 28 ARG HA H 4.16 0.03 1 89 . 28 ARG CA C 57.67 0.05 1 90 . 28 ARG N N 127.67 0.05 1 91 . 30 ILE H H 7.75 0.03 1 92 . 30 ILE C C 176.01 0.05 1 93 . 30 ILE CA C 61.60 0.05 1 94 . 30 ILE N N 127.03 0.05 1 95 . 31 ALA H H 8.47 0.03 1 96 . 31 ALA HA H 4.77 0.03 1 97 . 31 ALA C C 177.55 0.05 1 98 . 31 ALA CA C 52.74 0.05 1 99 . 31 ALA N N 127.49 0.05 1 100 . 32 ASP H H 8.26 0.03 1 101 . 32 ASP C C 176.91 0.05 1 102 . 32 ASP CA C 54.45 0.05 1 103 . 32 ASP N N 119.28 0.05 1 104 . 33 GLU H H 8.54 0.03 1 105 . 33 GLU HA H 4.20 0.03 1 106 . 33 GLU C C 176.95 0.05 1 107 . 33 GLU CA C 57.13 0.05 1 108 . 33 GLU N N 121.02 0.05 1 109 . 34 ASN H H 8.47 0.03 1 110 . 34 ASN C C 176.08 0.05 1 111 . 34 ASN CA C 53.80 0.05 1 112 . 34 ASN N N 118.47 0.05 1 113 . 35 GLY H H 8.34 0.03 1 114 . 35 GLY HA2 H 3.90 0.03 1 115 . 35 GLY HA3 H 3.90 0.03 1 116 . 35 GLY C C 174.14 0.05 1 117 . 35 GLY CA C 45.72 0.05 1 118 . 35 GLY N N 108.39 0.05 1 119 . 36 LYS H H 7.98 0.03 1 120 . 36 LYS CA C 54.25 0.05 1 121 . 36 LYS N N 121.56 0.05 1 122 . 37 PRO C C 177.04 0.05 1 123 . 37 PRO CA C 63.07 0.05 1 124 . 38 ARG H H 8.48 0.03 1 125 . 38 ARG HA H 4.58 0.03 1 126 . 38 ARG CA C 54.07 0.05 1 127 . 38 ARG N N 122.62 0.05 1 128 . 39 PRO C C 176.76 0.05 1 129 . 39 PRO CA C 63.07 0.05 1 130 . 40 ALA H H 8.36 0.03 1 131 . 40 ALA HA H 4.20 0.03 1 132 . 40 ALA C C 177.78 0.05 1 133 . 40 ALA CA C 52.67 0.05 1 134 . 40 ALA N N 123.60 0.05 1 135 . 41 PHE H H 8.08 0.03 1 136 . 41 PHE HA H 4.63 0.03 1 137 . 41 PHE C C 175.96 0.05 1 138 . 41 PHE CA C 57.67 0.05 1 139 . 41 PHE N N 117.74 0.05 1 140 . 42 SER H H 8.18 0.03 1 141 . 42 SER HA H 4.36 0.03 1 142 . 42 SER C C 176.89 0.05 1 143 . 42 SER CA C 58.17 0.05 1 144 . 42 SER N N 116.32 0.05 1 145 . 43 ASN H H 8.39 0.03 1 146 . 43 ASN C C 175.09 0.05 1 147 . 43 ASN CA C 53.37 0.05 1 148 . 43 ASN N N 120.54 0.05 1 149 . 44 ALA H H 8.19 0.03 1 150 . 44 ALA HA H 4.74 0.03 1 151 . 44 ALA C C 176.19 0.05 1 152 . 44 ALA CA C 53.57 0.05 1 153 . 44 ALA N N 123.48 0.05 1 154 . 45 LEU H H 8.12 0.03 1 155 . 45 LEU C C 177.42 0.05 1 156 . 45 LEU N N 120.40 0.05 1 157 . 46 GLU H H 8.19 0.03 1 158 . 46 GLU HA H 4.53 0.03 1 159 . 46 GLU CA C 54.57 0.05 1 160 . 46 GLU N N 122.29 0.05 1 161 . 47 PRO C C 176.76 0.05 1 162 . 47 PRO CA C 63.37 0.05 1 163 . 48 ASP H H 8.30 0.03 1 164 . 48 ASP HA H 4.53 0.03 1 165 . 48 ASP C C 176.60 0.05 1 166 . 48 ASP CA C 54.37 0.05 1 167 . 48 ASP N N 119.01 0.05 1 168 . 49 PHE H H 8.08 0.03 1 169 . 49 PHE CA C 57.87 0.05 1 170 . 49 PHE N N 120.62 0.05 1 171 . 50 LYS N N 127.48 0.05 1 172 . 53 SER N N 117.35 0.05 1 173 . 54 ILE H H 8.18 0.03 1 174 . 54 ILE N N 122.12 0.05 1 175 . 55 ALA H H 8.34 0.03 1 176 . 55 ALA C C 177.95 0.05 1 177 . 55 ALA N N 127.41 0.05 1 178 . 56 VAL H H 8.12 0.03 1 179 . 56 VAL HA H 4.17 0.03 1 180 . 56 VAL C C 176.80 0.05 1 181 . 56 VAL CA C 62.37 0.05 1 182 . 56 VAL N N 118.47 0.05 1 183 . 57 THR H H 8.19 0.03 1 184 . 57 THR HA H 4.35 0.03 1 185 . 57 THR C C 175.44 0.05 1 186 . 57 THR CA C 62.17 0.05 1 187 . 57 THR N N 116.63 0.05 1 188 . 58 GLY H H 8.42 0.03 1 189 . 58 GLY HA2 H 4.00 0.03 1 190 . 58 GLY HA3 H 4.00 0.03 1 191 . 58 GLY N N 110.62 0.05 1 192 . 59 SER H H 8.31 0.03 1 193 . 59 SER HA H 4.43 0.03 1 194 . 59 SER C C 175.45 0.05 1 195 . 59 SER CA C 58.70 0.05 1 196 . 59 SER N N 115.22 0.05 1 197 . 60 GLY H H 8.55 0.03 1 198 . 60 GLY HA2 H 3.97 0.03 1 199 . 60 GLY HA3 H 3.97 0.03 1 200 . 60 GLY N N 110.58 0.05 1 201 . 61 SER H H 8.26 0.03 1 202 . 61 SER HA H 4.43 0.03 1 203 . 61 SER CA C 58.58 0.05 1 204 . 61 SER N N 115.26 0.05 1 205 . 64 HIS C C 175.08 0.05 1 206 . 65 LEU H H 8.16 0.03 1 207 . 65 LEU C C 176.96 0.05 1 208 . 65 LEU CA C 55.59 0.05 1 209 . 65 LEU N N 122.85 0.05 1 210 . 66 GLN H H 8.31 0.03 1 211 . 66 GLN HA H 4.30 0.03 1 212 . 66 GLN C C 175.35 0.05 1 213 . 66 GLN CA C 55.78 0.05 1 214 . 66 GLN N N 120.27 0.05 1 215 . 67 PHE H H 8.23 0.03 1 216 . 67 PHE HA H 4.64 0.03 1 217 . 67 PHE C C 175.08 0.05 1 218 . 67 PHE CA C 57.57 0.05 1 219 . 67 PHE N N 121.27 0.05 1 220 . 68 ILE H H 8.08 0.03 1 221 . 68 ILE HA H 4.39 0.03 1 222 . 68 ILE CA C 58.07 0.05 1 223 . 68 ILE N N 125.00 0.05 1 224 . 69 PRO C C 176.18 0.05 1 225 . 70 VAL H H 8.16 0.03 1 226 . 70 VAL HA H 4.01 0.03 1 227 . 70 VAL C C 175.88 0.05 1 228 . 70 VAL CA C 62.17 0.05 1 229 . 70 VAL N N 120.09 0.05 1 230 . 71 ALA H H 8.34 0.03 1 231 . 71 ALA HA H 4.58 0.03 1 232 . 71 ALA CA C 50.47 0.05 1 233 . 71 ALA N N 129.03 0.05 1 234 . 73 PRO C C 177.06 0.05 1 235 . 74 SER H H 8.43 0.03 1 236 . 74 SER HA H 4.48 0.03 1 237 . 74 SER CA C 56.47 0.05 1 238 . 74 SER N N 117.01 0.05 1 239 . 75 PRO C C 177.93 0.05 1 240 . 75 PRO CA C 63.67 0.05 1 241 . 76 GLY H H 8.50 0.03 1 242 . 76 GLY HA2 H 3.96 0.03 1 243 . 76 GLY HA3 H 3.96 0.03 1 244 . 76 GLY C C 174.66 0.05 1 245 . 76 GLY CA C 45.57 0.05 1 246 . 76 GLY N N 109.17 0.05 1 247 . 77 SER H H 8.15 0.03 1 248 . 77 SER HA H 4.44 0.03 1 249 . 77 SER C C 174.98 0.05 1 250 . 77 SER CA C 58.47 0.05 1 251 . 77 SER N N 115.28 0.05 1 252 . 78 SER H H 8.39 0.03 1 253 . 78 SER C C 174.23 0.05 1 254 . 78 SER CA C 58.47 0.05 1 255 . 78 SER N N 117.40 0.05 1 256 . 79 ALA H H 8.25 0.03 1 257 . 79 ALA C C 177.23 0.05 1 258 . 79 ALA CA C 52.37 0.05 1 259 . 79 ALA N N 125.47 0.05 1 260 . 80 ALA H H 8.23 0.03 1 261 . 80 ALA HA H 4.55 0.03 1 262 . 80 ALA CA C 50.67 0.05 1 263 . 80 ALA N N 124.50 0.05 1 264 . 81 PRO C C 176.95 0.05 1 265 . 81 PRO CA C 62.97 0.05 1 266 . 82 ALA H H 8.46 0.03 1 267 . 82 ALA HA H 4.34 0.03 1 268 . 82 ALA C C 178.17 0.05 1 269 . 82 ALA CA C 52.67 0.05 1 270 . 82 ALA N N 124.23 0.05 1 271 . 83 THR H H 8.08 0.03 1 272 . 83 THR HA H 4.27 0.03 1 273 . 83 THR C C 174.66 0.05 1 274 . 83 THR CA C 61.87 0.05 1 275 . 83 THR N N 112.84 0.05 1 276 . 84 GLU H H 8.39 0.03 1 277 . 84 GLU HA H 4.30 0.03 1 278 . 84 GLU C C 176.33 0.05 1 279 . 84 GLU CA C 56.47 0.05 1 280 . 84 GLU N N 123.17 0.05 1 281 . 85 VAL H H 8.28 0.03 1 282 . 85 VAL HA H 4.39 0.03 1 283 . 85 VAL CA C 59.87 0.05 1 284 . 85 VAL N N 122.64 0.05 1 285 . 86 PRO C C 176.89 0.05 1 286 . 86 PRO CA C 63.17 0.05 1 287 . 87 ASP H H 8.38 0.03 1 288 . 87 ASP HA H 4.51 0.03 1 289 . 87 ASP C C 176.20 0.05 1 290 . 87 ASP CA C 54.57 0.05 1 291 . 87 ASP N N 120.12 0.05 1 292 . 88 ARG H H 8.12 0.03 1 293 . 88 ARG C C 175.86 0.05 1 294 . 88 ARG CA C 55.77 0.05 1 295 . 88 ARG N N 120.28 0.05 1 296 . 89 ASP H H 8.51 0.03 1 297 . 89 ASP HA H 4.81 0.03 1 298 . 89 ASP CA C 52.67 0.05 1 299 . 89 ASP N N 123.57 0.05 1 300 . 90 PRO C C 177.53 0.05 1 301 . 90 PRO CA C 63.57 0.05 1 302 . 91 GLU H H 8.49 0.03 1 303 . 91 GLU HA H 4.20 0.03 1 304 . 91 GLU C C 176.95 0.05 1 305 . 91 GLU CA C 56.87 0.05 1 306 . 91 GLU N N 119.41 0.05 1 307 . 92 LYS H H 8.09 0.03 1 308 . 92 LYS HA H 4.11 0.03 1 309 . 92 LYS C C 175.02 0.05 1 310 . 92 LYS CA C 56.17 0.05 1 311 . 92 LYS N N 121.47 0.05 1 312 . 93 SER H H 8.43 0.03 1 313 . 93 SER C C 176.77 0.05 1 314 . 93 SER CA C 58.47 0.05 1 315 . 93 SER N N 117.48 0.05 1 316 . 94 SER H H 8.46 0.03 1 317 . 94 SER HA H 4.48 0.03 1 318 . 94 SER C C 175.08 0.05 1 319 . 94 SER CA C 58.57 0.05 1 320 . 94 SER N N 117.72 0.05 1 321 . 95 GLU H H 8.50 0.03 1 322 . 95 GLU HA H 4.28 0.03 1 323 . 95 GLU C C 176.49 0.05 1 324 . 95 GLU CA C 56.97 0.05 1 325 . 95 GLU N N 122.07 0.05 1 326 . 96 ASP H H 8.19 0.03 1 327 . 96 ASP C C 176.49 0.05 1 328 . 96 ASP CA C 54.67 0.05 1 329 . 96 ASP N N 119.84 0.05 1 330 . 97 ASP H H 8.17 0.03 1 331 . 97 ASP C C 175.92 0.05 1 332 . 97 ASP CA C 54.57 0.05 1 333 . 97 ASP N N 120.14 0.05 1 334 . 98 VAL H H 7.87 0.03 1 335 . 98 VAL HA H 4.03 0.03 1 336 . 98 VAL C C 175.17 0.05 1 337 . 98 VAL CA C 62.27 0.05 1 338 . 98 VAL N N 119.10 0.05 1 339 . 99 TYR H H 7.73 0.03 1 340 . 99 TYR HA H 4.38 0.03 1 341 . 99 TYR CA C 59.27 0.05 1 342 . 99 TYR N N 128.36 0.05 1 stop_ save_