data_5948 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Structure Analysis of Integrin alpha IIb beta 3 - Specific Disintegrin with the AKGDWN Motif ; _BMRB_accession_number 5948 _BMRB_flat_file_name bmr5948.str _Entry_type original _Submission_date 2003-09-16 _Accession_date 2003-09-16 _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 Chuang Woei-Jer . . 2 Chen Chiu-Yueh . . 3 Shiu Jia-Hau . . 4 Liu Yu-Chen . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 345 "15N chemical shifts" 64 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2006-01-12 original author . stop_ loop_ _Related_BMRB_accession_number _Relationship 5949 'Disintegrin kistrin, ARGDWN Mutant' stop_ _Original_release_date 2006-01-12 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Structure Analysis of Integrin alpha IIb beta 3 - Specific Disintegrin with the AKGDWN Motif ; _Citation_status 'in preparation' _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Chuang Woei-Jer . . 2 Chen Chiu-Yueh . . 3 Shiu Jia-Hau . . 4 Liu Yu-Chen . . stop_ _Journal_abbreviation . _Journal_volume . _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year . _Details . loop_ _Keyword disintegrin integrin NMR rhodostomin stop_ save_ ####################################### # Cited references within the entry # ####################################### save_ref_1 _Saveframe_category citation _Citation_full ; Guo RT, Chou LJ, Chen YC, Chen CY, Pari K, Jen CJ, Lo SJ, Huang SL, Lee CY, Chang TW, Chaung WJ. Expression in Pichia pastoris and characterization by circular dichroism and NMR of rhodostomin. Proteins. 2001 Jun 1;43(4):499-508. ; _Citation_title 'Expression in Pichia pastoris and characterization by circular dichroism and NMR of rhodostomin.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 11340665 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Guo 'R T' T. . 2 Chou 'L J' J. . 3 Chen 'Y C' C. . 4 Chen 'C Y' Y. . 5 Pari K . . 6 Jen 'C J' J. . 7 Lo 'S J' J. . 8 Huang 'S L' L. . 9 Lee 'C Y' Y. . 10 Chang 'T W' W. . 11 Chaung 'W J' J. . stop_ _Journal_abbreviation Proteins _Journal_name_full Proteins _Journal_volume 43 _Journal_issue 4 _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 499 _Page_last 508 _Year 2001 _Details ; Rhodostomin (Rho) is a snake venom protein isolated from Calloselasma rhodostoma. Rho is a disintegrin that inhibits platelet aggregation by blocking the binding of fibrinogen to the integrin alpha(IIb)beta3 of platelets. Rho produced in Escherichia coli inhibited platelet aggregation with a K(I) value of 263 nM. Although functional, Rho produced in E. coli is misfolded based on our 2D and 3D NMR studies. In order to correct the folding problem, Rho was expressed in Pichia pastoris. The recombinant Rho expressed in P. pastoris inhibited platelet aggregation with a resulting K(I) value of 70 nM. This is the same potency as that of native Rho. CD analysis showed that the secondary structures of Rho are pH-independent and contain 3.5-7.9% alpha-helix, 48.2-50.5% beta-structures, and 42.3-47% coil. The sequential assignment and structure analysis of Rho were obtained using 2D and 3D 15N-edited NMR spectra. These results provide the first direct evidence that highly disulfide-bonded disintegrin can be expressed in P. pastoris with the correct fold. This evidence may serve as the basis for exploring the structure and function relationships as well as the dynamics of disintegrin and its variants. ; save_ ################################## # Molecular system description # ################################## save_system_Rho _Saveframe_category molecular_system _Mol_system_name 'Disintegrin kistrin, AKGDWN Mutant' _Abbreviation_common Rho _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Disintegrin kistrin' $Rho_monomer stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Rho_monomer _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Rhodostomin _Name_variant P48A,M52W,P53N _Abbreviation_common Rho _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 68 _Mol_residue_sequence ; GKECDCSSPENPCCDAATCK LRPGAQCGEGLCCEQCKFSR AGKICRIAKGDWNDDRCTGQ SADCPRYH ; loop_ _Residue_seq_code _Residue_label 1 GLY 2 LYS 3 GLU 4 CYS 5 ASP 6 CYS 7 SER 8 SER 9 PRO 10 GLU 11 ASN 12 PRO 13 CYS 14 CYS 15 ASP 16 ALA 17 ALA 18 THR 19 CYS 20 LYS 21 LEU 22 ARG 23 PRO 24 GLY 25 ALA 26 GLN 27 CYS 28 GLY 29 GLU 30 GLY 31 LEU 32 CYS 33 CYS 34 GLU 35 GLN 36 CYS 37 LYS 38 PHE 39 SER 40 ARG 41 ALA 42 GLY 43 LYS 44 ILE 45 CYS 46 ARG 47 ILE 48 ALA 49 LYS 50 GLY 51 ASP 52 TRP 53 ASN 54 ASP 55 ASP 56 ARG 57 CYS 58 THR 59 GLY 60 GLN 61 SER 62 ALA 63 ASP 64 CYS 65 PRO 66 ARG 67 TYR 68 HIS 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 BMRB 19210 rhodostomin 100.00 76 98.53 100.00 9.15e-40 BMRB 19211 rhodostomin 97.06 79 98.48 100.00 1.73e-37 BMRB 19212 rhodostomin 97.06 79 98.48 100.00 3.05e-37 BMRB 5949 Rho_monomer 100.00 68 98.53 100.00 2.04e-39 PDB 1Q7I "Structural Analysis Of Integrin Alpha Iib Beta 3- Disintegrin With The Akgdwn Motif" 100.00 68 98.53 100.00 2.04e-39 PDB 1Q7J "Structural Analysis Of Integrin Alpha Iib Beta 3- Disintegrin With The Akgdwn Motif" 100.00 68 100.00 100.00 8.53e-40 PDB 2M75 "The C-terminal Region Of Disintegrin Modulate Its 3d Conformation And Cooperate With Rgd Loop In Regulating Recognitions Of Int" 100.00 76 98.53 100.00 9.15e-40 PDB 2M7F "The C-terminal Region Of Disintegrin Modulate Its 3d Conformation And Cooperate With Rgd Loop In Regulating Integrins Recogniti" 97.06 79 98.48 100.00 1.73e-37 PDB 2M7H "The C-terminal Region Of Disintegrin Modulate Its 3d Conformation And Cooperate With Rgd Loop In Regulating Integrin Alpha-iib " 97.06 79 98.48 100.00 3.05e-37 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $Rho_monomer 'Calloselasma rhodostoma' 8717 Eukaryota Metazoa Calloselasma rhodostoma stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Vector_type _Vector_name $Rho_monomer 'recombinant technology' 'P. pastoris' Pichia pastoris X33 plasmid pPICZa 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 _Isotopic_labeling $Rho_monomer 3 mM . H2O 90 % . D2O 10 % . stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Rho_monomer 3 mM . D2O 100 % . stop_ save_ save_sample_3 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Rho_monomer 2.5 mM [U-15N] H2O 90 % . D2O 10 % . stop_ save_ ############################ # Computer software used # ############################ save_XWINNMR _Saveframe_category software _Name xwinnmr _Version 2.6 loop_ _Task processing stop_ _Details Bruker save_ save_AURELIA _Saveframe_category software _Name AURELIA _Version 2.7.10 loop_ _Task 'data analysis' stop_ _Details Neidig save_ save_X-PLOR _Saveframe_category software _Name X-PLOR _Version 3.85 loop_ _Task refinement stop_ _Details Brunger save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model AVANCE _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_NOESY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D NOESY' _Sample_label . save_ save_2D_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D TOCSY' _Sample_label . save_ save_3D_15N-separated_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 15N-separated NOESY' _Sample_label . save_ save_3D_15N-separated_TOCSY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 15N-separated TOCSY' _Sample_label . save_ save_HNHA_5 _Saveframe_category NMR_applied_experiment _Experiment_name HNHA _Sample_label . save_ save_15N-HSQC_6 _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 15N-separated NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 15N-separated TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_5 _Saveframe_category NMR_applied_experiment _Experiment_name HNHA _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_6 _Saveframe_category NMR_applied_experiment _Experiment_name 15N-HSQC _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_sample_cond_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units 'ionic strength' 0 . M pH 6.0 0.01 pH pressure 1 . atm temperature 300 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 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_chemical_shift_set_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D NOESY' '2D TOCSY' '3D 15N-separated NOESY' '3D 15N-separated TOCSY' HNHA 15N-HSQC stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_cond_1 _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name 'Disintegrin kistrin' _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 . 1 GLY H H 8.56 0.01 1 2 . 1 GLY HA2 H 4.68 0.01 2 3 . 1 GLY HA3 H 4.01 0.01 2 4 . 1 GLY N N 114.4 0.1 1 5 . 2 LYS H H 8.33 0.01 1 6 . 2 LYS HB2 H 1.90 0.01 2 7 . 2 LYS HB3 H 1.81 0.01 2 8 . 2 LYS HG2 H 1.48 0.01 1 9 . 2 LYS HG3 H 1.48 0.01 1 10 . 2 LYS N N 125.3 0.1 1 11 . 3 GLU H H 8.66 0.01 1 12 . 3 GLU HA H 4.32 0.01 1 13 . 3 GLU HB2 H 2.05 0.01 2 14 . 3 GLU HB3 H 1.97 0.01 2 15 . 3 GLU HG2 H 2.28 0.01 1 16 . 3 GLU HG3 H 2.28 0.01 1 17 . 3 GLU N N 125.4 0.1 1 18 . 4 CYS H H 8.12 0.01 1 19 . 4 CYS HA H 4.98 0.01 1 20 . 4 CYS HB2 H 3.46 0.01 2 21 . 4 CYS HB3 H 2.58 0.01 2 22 . 4 CYS N N 123.8 0.1 1 23 . 5 ASP H H 8.78 0.01 1 24 . 5 ASP HA H 4.82 0.01 1 25 . 5 ASP HB2 H 2.72 0.01 1 26 . 5 ASP HB3 H 2.72 0.01 1 27 . 5 ASP N N 123.8 0.1 1 28 . 6 CYS H H 6.98 0.01 1 29 . 6 CYS HA H 4.93 0.01 1 30 . 6 CYS HB2 H 3.22 0.01 2 31 . 6 CYS HB3 H 2.99 0.01 2 32 . 6 CYS N N 116.7 0.1 1 33 . 7 SER H H 9.66 0.01 1 34 . 7 SER HA H 4.23 0.01 1 35 . 7 SER HB2 H 3.95 0.01 2 36 . 7 SER HB3 H 3.86 0.01 2 37 . 7 SER N N 123.5 0.1 1 38 . 8 SER H H 7.86 0.01 1 39 . 8 SER HA H 5.14 0.01 1 40 . 8 SER HB2 H 4.01 0.01 2 41 . 8 SER HB3 H 3.82 0.01 2 42 . 8 SER N N 119.7 0.1 1 43 . 9 PRO HA H 4.56 0.01 1 44 . 9 PRO HB2 H 2.39 0.01 1 45 . 9 PRO HB3 H 2.39 0.01 1 46 . 9 PRO HG2 H 2.04 0.01 1 47 . 9 PRO HG3 H 2.04 0.01 1 48 . 9 PRO HD2 H 3.97 0.01 2 49 . 9 PRO HD3 H 3.82 0.01 2 50 . 10 GLU H H 8.24 0.01 1 51 . 10 GLU HA H 4.29 0.01 1 52 . 10 GLU HB2 H 2.20 0.01 2 53 . 10 GLU HB3 H 2.03 0.01 2 54 . 10 GLU HG2 H 2.26 0.01 1 55 . 10 GLU HG3 H 2.26 0.01 1 56 . 10 GLU N N 120.9 0.1 1 57 . 11 ASN H H 7.26 0.01 1 58 . 11 ASN HA H 4.79 0.01 1 59 . 11 ASN HB2 H 3.15 0.01 2 60 . 11 ASN HB3 H 2.87 0.01 2 61 . 11 ASN HD21 H 8.30 0.01 2 62 . 11 ASN HD22 H 7.44 0.01 2 63 . 11 ASN N N 124.8 0.1 1 64 . 12 PRO HA H 4.70 0.01 1 65 . 12 PRO HB2 H 2.39 0.01 2 66 . 12 PRO HB3 H 1.98 0.01 2 67 . 12 PRO HG2 H 2.15 0.01 1 68 . 12 PRO HG3 H 2.15 0.01 1 69 . 12 PRO HD2 H 4.17 0.01 2 70 . 12 PRO HD3 H 3.97 0.01 2 71 . 13 CYS H H 8.93 0.01 1 72 . 13 CYS HA H 4.65 0.01 1 73 . 13 CYS HB2 H 3.22 0.01 2 74 . 13 CYS HB3 H 2.58 0.01 2 75 . 13 CYS N N 121.9 0.1 1 76 . 14 CYS H H 7.54 0.01 1 77 . 14 CYS HA H 5.06 0.01 1 78 . 14 CYS HB2 H 2.95 0.01 2 79 . 14 CYS HB3 H 2.89 0.01 2 80 . 14 CYS N N 116.5 0.1 1 81 . 15 ASP H H 8.21 0.01 1 82 . 15 ASP HA H 4.57 0.01 1 83 . 15 ASP HB2 H 3.14 0.01 2 84 . 15 ASP HB3 H 2.48 0.01 2 85 . 15 ASP N N 125.2 0.1 1 86 . 16 ALA H H 9.00 0.01 1 87 . 16 ALA HA H 4.15 0.01 1 88 . 16 ALA HB H 1.55 0.01 1 89 . 16 ALA N N 135.3 0.1 1 90 . 17 ALA H H 8.67 0.01 1 91 . 17 ALA HA H 4.36 0.01 1 92 . 17 ALA HB H 1.55 0.01 1 93 . 17 ALA N N 121.9 0.1 1 94 . 18 THR H H 7.74 0.01 1 95 . 18 THR HA H 4.45 0.01 1 96 . 18 THR HB H 4.34 0.01 1 97 . 18 THR HG2 H 1.20 0.01 1 98 . 18 THR N N 109.5 0.1 1 99 . 19 CYS H H 8.60 0.01 1 100 . 19 CYS HA H 4.64 0.01 1 101 . 19 CYS HB2 H 3.54 0.01 2 102 . 19 CYS HB3 H 3.32 0.01 2 103 . 19 CYS N N 123.2 0.1 1 104 . 20 LYS H H 7.96 0.01 1 105 . 20 LYS HA H 4.95 0.01 1 106 . 20 LYS HB2 H 1.98 0.01 2 107 . 20 LYS HB3 H 1.54 0.01 2 108 . 20 LYS HG2 H 1.68 0.01 1 109 . 20 LYS HG3 H 1.68 0.01 1 110 . 20 LYS N N 122.5 0.1 1 111 . 21 LEU H H 9.21 0.01 1 112 . 21 LEU HA H 4.29 0.01 1 113 . 21 LEU HB2 H 1.42 0.01 1 114 . 21 LEU HB3 H 1.42 0.01 1 115 . 21 LEU HG H 1.68 0.01 1 116 . 21 LEU HD1 H 0.63 0.01 2 117 . 21 LEU HD2 H 0.52 0.01 2 118 . 21 LEU N N 124.6 0.1 1 119 . 22 ARG H H 8.49 0.01 1 120 . 22 ARG HA H 4.45 0.01 1 121 . 22 ARG HB2 H 1.72 0.01 2 122 . 22 ARG HB3 H 1.63 0.01 2 123 . 22 ARG HD2 H 3.25 0.01 1 124 . 22 ARG HD3 H 3.25 0.01 1 125 . 22 ARG N N 126.6 0.1 1 126 . 23 PRO HA H 4.39 0.01 1 127 . 23 PRO HB2 H 2.38 0.01 2 128 . 23 PRO HB3 H 1.92 0.01 2 129 . 23 PRO HG2 H 2.09 0.01 1 130 . 23 PRO HG3 H 2.09 0.01 1 131 . 24 GLY H H 8.83 0.01 1 132 . 24 GLY HA2 H 4.30 0.01 2 133 . 24 GLY HA3 H 3.66 0.01 2 134 . 24 GLY N N 117.7 0.1 1 135 . 25 ALA H H 7.94 0.01 1 136 . 25 ALA HA H 4.54 0.01 1 137 . 25 ALA HB H 1.62 0.01 1 138 . 25 ALA N N 127.6 0.1 1 139 . 26 GLN H H 8.83 0.01 1 140 . 26 GLN HA H 4.11 0.01 1 141 . 26 GLN HB2 H 1.24 0.01 2 142 . 26 GLN HB3 H 0.57 0.01 2 143 . 26 GLN HG2 H 2.17 0.01 2 144 . 26 GLN HG3 H 2.06 0.01 2 145 . 26 GLN HE21 H 6.99 0.01 2 146 . 26 GLN HE22 H 7.72 0.01 2 147 . 26 GLN N N 122.1 0.1 1 148 . 27 CYS H H 7.81 0.01 1 149 . 27 CYS HA H 4.56 0.01 1 150 . 27 CYS HB2 H 3.29 0.01 2 151 . 27 CYS HB3 H 3.15 0.01 2 152 . 27 CYS N N 117.4 0.1 1 153 . 28 GLY H H 9.16 0.01 1 154 . 28 GLY HA2 H 4.45 0.01 2 155 . 28 GLY HA3 H 3.57 0.01 2 156 . 28 GLY N N 109.0 0.1 1 157 . 29 GLU H H 7.84 0.01 1 158 . 29 GLU HA H 4.78 0.01 1 159 . 29 GLU HB2 H 2.31 0.01 2 160 . 29 GLU HB3 H 1.98 0.01 2 161 . 29 GLU HG2 H 2.26 0.01 2 162 . 29 GLU HG3 H 2.17 0.01 2 163 . 29 GLU N N 122.2 0.1 1 164 . 30 GLY H H 8.39 0.01 1 165 . 30 GLY HA2 H 4.92 0.01 2 166 . 30 GLY HA3 H 3.93 0.01 2 167 . 30 GLY N N 112.7 0.1 1 168 . 31 LEU H H 8.96 0.01 1 169 . 31 LEU HA H 4.35 0.01 1 170 . 31 LEU HB2 H 1.83 0.01 2 171 . 31 LEU HB3 H 1.74 0.01 2 172 . 31 LEU HG H 1.99 0.01 1 173 . 31 LEU HD1 H 1.14 0.01 1 174 . 31 LEU HD2 H 1.14 0.01 1 175 . 31 LEU N N 125.0 0.1 1 176 . 32 CYS H H 8.47 0.01 1 177 . 32 CYS HA H 5.11 0.01 1 178 . 32 CYS HB2 H 3.89 0.01 2 179 . 32 CYS HB3 H 2.51 0.01 2 180 . 32 CYS N N 119.2 0.1 1 181 . 33 CYS H H 7.37 0.01 1 182 . 33 CYS HA H 5.29 0.01 1 183 . 33 CYS HB2 H 2.99 0.01 2 184 . 33 CYS HB3 H 2.56 0.01 2 185 . 33 CYS N N 125.3 0.1 1 186 . 34 GLU H H 9.74 0.01 1 187 . 34 GLU HA H 4.68 0.01 1 188 . 34 GLU HB2 H 2.06 0.01 2 189 . 34 GLU HB3 H 1.91 0.01 2 190 . 34 GLU HG2 H 2.23 0.01 1 191 . 34 GLU HG3 H 2.23 0.01 1 192 . 34 GLU N N 132.7 0.1 1 193 . 35 GLN H H 9.45 0.01 1 194 . 35 GLN HA H 4.00 0.01 1 195 . 35 GLN HB2 H 2.40 0.01 2 196 . 35 GLN HB3 H 2.13 0.01 2 197 . 35 GLN HG2 H 2.24 0.01 1 198 . 35 GLN HG3 H 2.24 0.01 1 199 . 35 GLN N N 129.6 0.1 1 200 . 36 CYS H H 8.49 0.01 1 201 . 36 CYS HA H 4.80 0.01 1 202 . 36 CYS HB2 H 3.76 0.01 2 203 . 36 CYS HB3 H 3.37 0.01 2 204 . 36 CYS N N 109.6 0.1 1 205 . 37 LYS H H 8.02 0.01 1 206 . 37 LYS HA H 4.70 0.01 1 207 . 37 LYS HB2 H 1.88 0.01 2 208 . 37 LYS HB3 H 1.79 0.01 2 209 . 37 LYS HG2 H 1.54 0.01 2 210 . 37 LYS HG3 H 1.46 0.01 2 211 . 37 LYS HD2 H 1.64 0.01 1 212 . 37 LYS HD3 H 1.64 0.01 1 213 . 37 LYS HE2 H 3.04 0.01 1 214 . 37 LYS HE3 H 3.04 0.01 1 215 . 37 LYS N N 125.1 0.1 1 216 . 38 PHE H H 8.28 0.01 1 217 . 38 PHE HA H 5.14 0.01 1 218 . 38 PHE HB2 H 3.05 0.01 2 219 . 38 PHE HB3 H 2.72 0.01 2 220 . 38 PHE HD1 H 7.03 0.01 1 221 . 38 PHE HD2 H 7.03 0.01 1 222 . 38 PHE HE1 H 7.26 0.01 1 223 . 38 PHE HE2 H 7.26 0.01 1 224 . 38 PHE N N 123.9 0.1 1 225 . 39 SER H H 9.04 0.01 1 226 . 39 SER HA H 4.27 0.01 1 227 . 39 SER HB2 H 3.93 0.01 2 228 . 39 SER HB3 H 3.77 0.01 2 229 . 39 SER N N 125.7 0.1 1 230 . 40 ARG H H 8.57 0.01 1 231 . 40 ARG HA H 4.10 0.01 1 232 . 40 ARG HB2 H 1.82 0.01 1 233 . 40 ARG HB3 H 1.82 0.01 1 234 . 40 ARG HG2 H 1.69 0.01 1 235 . 40 ARG HG3 H 1.69 0.01 1 236 . 40 ARG HD2 H 3.27 0.01 1 237 . 40 ARG HD3 H 3.27 0.01 1 238 . 40 ARG N N 125.8 0.1 1 239 . 41 ALA H H 8.50 0.01 1 240 . 41 ALA HA H 3.72 0.01 1 241 . 41 ALA HB H 1.30 0.01 1 242 . 41 ALA N N 127.7 0.1 1 243 . 42 GLY H H 8.62 0.01 1 244 . 42 GLY HA2 H 4.49 0.01 2 245 . 42 GLY HA3 H 3.59 0.01 2 246 . 42 GLY N N 114.6 0.1 1 247 . 43 LYS H H 7.54 0.01 1 248 . 43 LYS HA H 4.15 0.01 1 249 . 43 LYS HB2 H 1.87 0.01 1 250 . 43 LYS HB3 H 1.87 0.01 1 251 . 43 LYS HG2 H 1.53 0.01 2 252 . 43 LYS HG3 H 1.42 0.01 2 253 . 43 LYS HD2 H 1.70 0.01 1 254 . 43 LYS HD3 H 1.70 0.01 1 255 . 43 LYS HE2 H 3.12 0.01 1 256 . 43 LYS HE3 H 3.12 0.01 1 257 . 43 LYS N N 126.8 0.1 1 258 . 44 ILE H H 8.90 0.01 1 259 . 44 ILE HA H 4.12 0.01 1 260 . 44 ILE HB H 1.85 0.01 1 261 . 44 ILE HG12 H 1.02 0.01 1 262 . 44 ILE HG13 H 1.02 0.01 1 263 . 44 ILE HD1 H 0.96 0.01 1 264 . 44 ILE N N 132.9 0.1 1 265 . 45 CYS H H 9.42 0.01 1 266 . 45 CYS HA H 5.19 0.01 1 267 . 45 CYS HB2 H 3.21 0.01 2 268 . 45 CYS HB3 H 3.04 0.01 2 269 . 45 CYS N N 127.9 0.1 1 270 . 46 ARG H H 7.61 0.01 1 271 . 46 ARG HA H 4.48 0.01 1 272 . 46 ARG HB2 H 2.11 0.01 1 273 . 46 ARG HB3 H 2.11 0.01 1 274 . 46 ARG HG2 H 1.76 0.01 2 275 . 46 ARG HG3 H 1.59 0.01 2 276 . 46 ARG N N 127.4 0.1 1 277 . 47 ILE H H 8.52 0.01 1 278 . 47 ILE HA H 4.29 0.01 1 279 . 47 ILE HB H 1.84 0.01 1 280 . 47 ILE HG12 H 1.16 0.01 1 281 . 47 ILE HG13 H 1.16 0.01 1 282 . 47 ILE HG2 H 1.03 0.01 1 283 . 47 ILE HD1 H 0.93 0.01 1 284 . 47 ILE N N 131.3 0.1 1 285 . 48 ALA H H 9.37 0.01 1 286 . 48 ALA HA H 4.32 0.01 1 287 . 48 ALA HB H 1.22 0.01 1 288 . 48 ALA N N 135.6 0.1 1 289 . 49 LYS H H 8.17 0.01 1 290 . 49 LYS HA H 4.41 0.01 1 291 . 49 LYS HB2 H 1.98 0.01 2 292 . 49 LYS HB3 H 1.82 0.01 2 293 . 49 LYS HG2 H 1.50 0.01 1 294 . 49 LYS HG3 H 1.50 0.01 1 295 . 49 LYS HD2 H 1.58 0.01 1 296 . 49 LYS HD3 H 1.58 0.01 1 297 . 49 LYS HE2 H 3.09 0.01 1 298 . 49 LYS HE3 H 3.09 0.01 1 299 . 49 LYS N N 125.3 0.1 1 300 . 50 GLY H H 7.40 0.01 1 301 . 50 GLY HA2 H 3.76 0.01 2 302 . 50 GLY HA3 H 3.27 0.01 2 303 . 50 GLY N N 113.5 0.1 1 304 . 51 ASP H H 8.14 0.01 1 305 . 51 ASP HA H 4.59 0.01 1 306 . 51 ASP HB2 H 2.66 0.01 2 307 . 51 ASP HB3 H 2.47 0.01 2 308 . 51 ASP N N 125.8 0.1 1 309 . 52 TRP H H 7.86 0.01 1 310 . 52 TRP HA H 4.80 0.01 1 311 . 52 TRP HB2 H 3.44 0.01 2 312 . 52 TRP HB3 H 3.27 0.01 2 313 . 52 TRP HD1 H 7.37 0.01 1 314 . 52 TRP HE1 H 10.22 0.01 1 315 . 52 TRP HE3 H 7.48 0.01 1 316 . 52 TRP HZ2 H 7.59 0.01 1 317 . 52 TRP HZ3 H 7.17 0.01 1 318 . 52 TRP HH2 H 7.23 0.01 1 319 . 52 TRP N N 123.6 0.1 1 320 . 53 ASN H H 8.33 0.01 1 321 . 53 ASN HA H 4.80 0.01 1 322 . 53 ASN HB2 H 2.59 0.01 2 323 . 53 ASN HB3 H 2.85 0.01 2 324 . 53 ASN HD21 H 6.99 0.01 2 325 . 53 ASN HD22 H 7.54 0.01 2 326 . 53 ASN N N 121.3 0.1 1 327 . 54 ASP H H 8.19 0.01 1 328 . 54 ASP HA H 4.70 0.01 1 329 . 54 ASP HB2 H 3.00 0.01 2 330 . 54 ASP HB3 H 2.41 0.01 2 331 . 54 ASP N N 124.1 0.1 1 332 . 55 ASP H H 8.64 0.01 1 333 . 55 ASP HA H 5.05 0.01 1 334 . 55 ASP HB2 H 3.03 0.01 2 335 . 55 ASP HB3 H 2.68 0.01 2 336 . 55 ASP N N 125.6 0.1 1 337 . 56 ARG H H 8.23 0.01 1 338 . 56 ARG HA H 5.34 0.01 1 339 . 56 ARG HB2 H 1.57 0.01 1 340 . 56 ARG HB3 H 1.57 0.01 1 341 . 56 ARG HG2 H 1.43 0.01 1 342 . 56 ARG HG3 H 1.43 0.01 1 343 . 56 ARG N N 124.0 0.1 1 344 . 57 CYS H H 9.34 0.01 1 345 . 57 CYS HA H 4.88 0.01 1 346 . 57 CYS HB2 H 3.92 0.01 2 347 . 57 CYS HB3 H 2.73 0.01 2 348 . 57 CYS N N 123.8 0.1 1 349 . 58 THR H H 10.18 0.01 1 350 . 58 THR HA H 4.44 0.01 1 351 . 58 THR HB H 4.51 0.01 1 352 . 58 THR HG2 H 1.42 0.01 1 353 . 58 THR N N 117.4 0.1 1 354 . 59 GLY H H 8.55 0.01 1 355 . 59 GLY HA2 H 4.46 0.01 1 356 . 59 GLY HA3 H 4.46 0.01 1 357 . 59 GLY N N 115.9 0.1 1 358 . 60 GLN H H 7.99 0.01 1 359 . 60 GLN HA H 4.30 0.01 1 360 . 60 GLN HB2 H 2.22 0.01 2 361 . 60 GLN HB3 H 1.69 0.01 2 362 . 60 GLN N N 119.0 0.1 1 363 . 61 SER H H 6.63 0.01 1 364 . 61 SER HA H 3.88 0.01 1 365 . 61 SER HB2 H 3.88 0.01 2 366 . 61 SER HB3 H 3.62 0.01 2 367 . 61 SER N N 114.9 0.1 1 368 . 62 ALA H H 9.03 0.01 1 369 . 62 ALA HA H 4.44 0.01 1 370 . 62 ALA HB H 1.83 0.01 1 371 . 62 ALA N N 129.6 0.1 1 372 . 63 ASP H H 8.08 0.01 1 373 . 63 ASP HA H 4.90 0.01 1 374 . 63 ASP HB2 H 2.67 0.01 2 375 . 63 ASP HB3 H 2.54 0.01 2 376 . 63 ASP N N 121.5 0.1 1 377 . 64 CYS H H 8.92 0.01 1 378 . 64 CYS HA H 5.40 0.01 1 379 . 64 CYS HB2 H 3.10 0.01 2 380 . 64 CYS HB3 H 2.80 0.01 2 381 . 64 CYS N N 125.9 0.1 1 382 . 65 PRO HA H 4.38 0.01 1 383 . 65 PRO HB2 H 2.22 0.01 2 384 . 65 PRO HB3 H 1.80 0.01 2 385 . 65 PRO HG2 H 2.13 0.01 2 386 . 65 PRO HG3 H 2.01 0.01 2 387 . 65 PRO HD2 H 3.99 0.01 2 388 . 65 PRO HD3 H 3.73 0.01 2 389 . 66 ARG H H 8.20 0.01 1 390 . 66 ARG HA H 4.36 0.01 1 391 . 66 ARG HB2 H 1.82 0.01 2 392 . 66 ARG HB3 H 1.76 0.01 2 393 . 66 ARG HG2 H 1.64 0.01 1 394 . 66 ARG HG3 H 1.64 0.01 1 395 . 66 ARG HD2 H 3.21 0.01 1 396 . 66 ARG HD3 H 3.21 0.01 1 397 . 66 ARG N N 123.6 0.1 1 398 . 67 TYR H H 8.65 0.01 1 399 . 67 TYR HA H 4.46 0.01 1 400 . 67 TYR HB2 H 3.14 0.01 2 401 . 67 TYR HB3 H 3.01 0.01 2 402 . 67 TYR HD1 H 7.16 0.01 2 403 . 67 TYR HD2 H 6.86 0.01 2 404 . 67 TYR N N 128.9 0.1 1 405 . 68 HIS H H 7.66 0.01 1 406 . 68 HIS HA H 4.37 0.01 1 407 . 68 HIS HB2 H 3.23 0.01 2 408 . 68 HIS HB3 H 3.09 0.01 2 409 . 68 HIS N N 129.3 0.1 1 stop_ save_