data_5117 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Structural and Dynamic Differences of Rhodostomin, an RGD-containing Disintegrin, and its D51E mutant ; _BMRB_accession_number 5117 _BMRB_flat_file_name bmr5117.str _Entry_type original _Submission_date 2001-08-20 _Accession_date 2001-08-20 _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 . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 350 "15N chemical shifts" 68 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2013-05-02 update BMRB 'update related entry loop' 2003-04-09 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 19210 'rhodostomin P48A/M52W/P53N mutant' 19211 'rhodostomin 48ARGDWN-67NGLYG mutant' 19212 'rhodostomin 48ARGDWN-67NPWNG mutant' stop_ save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Structural and Dynamic Differences of Rhodostomin, an RGD-containing Disintegrin, and its D51E mutant ; _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 RT . . 2 Chou LJ . . 3 Chen YC . . 4 Chen CY . . 5 Pari K . . 6 Jen CJ . . 7 Lo SJ . . 8 Huang SL . . 9 Lee CY . . 10 Chang TW . . 11 Chuang Woei-Jer . . stop_ _Journal_abbreviation 'Proteins: Struct., Funct., Genet.' _Journal_volume 43 _Journal_issue 4 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 499 _Page_last 508 _Year 2001 _Details . loop_ _Keyword dynamics expression 'Pichia pastoris' rhodostomin structure 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 Rhodostomin _Abbreviation_common Rho _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label Rhodostomin $Rho stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'all disulfide bound' loop_ _Biological_function 'integrin antagonist' stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_Rho _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Rhodostomin _Abbreviation_common Rho _Molecular_mass 8417 _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 76 _Mol_residue_sequence ; EFHHHHHHGKECDCSSPENP CCDAATCKLRPGAQCGEGLC CEQCKFSRAGKICRIPRGDM PDDRCTGQSADCPRYH ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 -7 GLU 2 -6 PHE 3 -5 HIS 4 -4 HIS 5 -3 HIS 6 -2 HIS 7 -1 HIS 8 0 HIS 9 1 GLY 10 2 LYS 11 3 GLU 12 4 CYS 13 5 ASP 14 6 CYS 15 7 SER 16 8 SER 17 9 PRO 18 10 GLU 19 11 ASN 20 12 PRO 21 13 CYS 22 14 CYS 23 15 ASP 24 16 ALA 25 17 ALA 26 18 THR 27 19 CYS 28 20 LYS 29 21 LEU 30 22 ARG 31 23 PRO 32 24 GLY 33 25 ALA 34 26 GLN 35 27 CYS 36 28 GLY 37 29 GLU 38 30 GLY 39 31 LEU 40 32 CYS 41 33 CYS 42 34 GLU 43 35 GLN 44 36 CYS 45 37 LYS 46 38 PHE 47 39 SER 48 40 ARG 49 41 ALA 50 42 GLY 51 43 LYS 52 44 ILE 53 45 CYS 54 46 ARG 55 47 ILE 56 48 PRO 57 49 ARG 58 50 GLY 59 51 ASP 60 52 MET 61 53 PRO 62 54 ASP 63 55 ASP 64 56 ARG 65 57 CYS 66 58 THR 67 59 GLY 68 60 GLN 69 61 SER 70 62 ALA 71 63 ASP 72 64 CYS 73 65 PRO 74 66 ARG 75 67 TYR 76 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 17963 Rhodostomin_G50L_mutant 89.47 68 98.53 98.53 1.90e-38 BMRB 2856 kistrin 89.47 68 100.00 100.00 1.60e-39 BMRB 7399 Rhodostoxin-disintegrin_rhodostomin 89.47 68 98.53 100.00 4.58e-39 BMRB 7400 Rhodostoxin-disintegrin_rhodostomin 89.47 68 98.53 98.53 9.92e-39 PDB 1N4Y "Refined Structure Of Kistrin" 89.47 68 100.00 100.00 1.60e-39 PDB 2LJV "Solution Structure Of Rhodostomin G50l Mutant" 89.47 68 98.53 98.53 1.90e-38 PDB 2PJF "Solution Structure Of Rhodostomin" 89.47 68 100.00 100.00 1.60e-39 PDB 2PJG "Solution Structure Of Rhodostomin D51e Mutant" 89.47 68 98.53 100.00 4.58e-39 PDB 2PJI "Solution Structure Of Rhodostomin P48a Mutant" 89.47 68 98.53 98.53 9.92e-39 PDB 4M4C "Crystal Structure Of Rhodostomin Argdp Mutant" 89.47 68 97.06 97.06 3.07e-37 GB AAA57568 "rhodostomin [synthetic construct]" 89.47 71 100.00 100.00 1.18e-39 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 Snake . Eukaryota Metazoa Calloslasma 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_name $Rho 'recombinant technology' Yeast Pichia pastoris X33 . 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 3.0 mM . stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $Rho 2.5 mM '[U-98% 15N]' stop_ save_ ############################ # Computer software used # ############################ save_xwinnmr _Saveframe_category software _Name xwinnmr _Version 2.6 loop_ _Task processing stop_ _Details . 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_1H-1H_DQF-COSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H DQF-COSY' _Sample_label . save_ save_2D_1H-1H_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label . save_ save_2D_1H-1H_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label . save_ save_2D_1H-15N_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label . save_ save_3D_1H-1H-15N_TOCSY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N TOCSY' _Sample_label . save_ save_3D_1H-1H-15N_NOESY_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N NOESY' _Sample_label . save_ save_3D_HNHA_7 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHA' _Sample_label . save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H DQF-COSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N TOCSY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_6 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-1H-15N NOESY' _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_7 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNHA' _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_condition _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 6.0 0.01 n/a 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 . . . 1.0 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_RGD6 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-1H DQF-COSY' '2D 1H-1H TOCSY' '2D 1H-1H NOESY' '2D 1H-15N HSQC' '3D 1H-1H-15N TOCSY' '3D 1H-1H-15N NOESY' '3D HNHA' stop_ _Sample_conditions_label $condition _Chem_shift_reference_set_label $chemical_shift_reference _Mol_system_component_name Rhodostomin _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 9 GLY H H 8.577 0.01 1 2 1 9 GLY HA2 H 4.678 0.01 2 3 1 9 GLY HA3 H 4.008 0.01 2 4 1 9 GLY N N 114.35 0.1 1 5 2 10 LYS H H 8.331 0.01 1 6 2 10 LYS HB2 H 1.90 0.01 2 7 2 10 LYS HB3 H 1.81 0.01 2 8 2 10 LYS HG2 H 1.484 0.01 4 9 2 10 LYS HG3 H 1.484 0.01 4 10 2 10 LYS N N 125.285 0.1 1 11 3 11 GLU H H 8.659 0.01 1 12 3 11 GLU HA H 4.322 0.01 1 13 3 11 GLU HB2 H 2.051 0.01 2 14 3 11 GLU HB3 H 1.970 0.01 2 15 3 11 GLU HG2 H 2.281 0.01 1 16 3 11 GLU HG3 H 2.281 0.01 1 17 3 11 GLU N N 125.356 0.1 1 18 4 12 CYS H H 8.116 0.01 1 19 4 12 CYS HA H 4.975 0.01 1 20 4 12 CYS HB2 H 3.456 0.01 2 21 4 12 CYS HB3 H 2.581 0.01 2 22 4 12 CYS N N 123.773 0.1 1 23 5 13 ASP H H 8.776 0.01 1 24 5 13 ASP HA H 4.817 0.01 1 25 5 13 ASP HB2 H 2.721 0.01 1 26 5 13 ASP HB3 H 2.721 0.01 1 27 5 13 ASP N N 123.756 0.1 1 28 6 14 CYS H H 6.975 0.01 1 29 6 14 CYS HA H 4.931 0.01 1 30 6 14 CYS HB2 H 3.218 0.01 2 31 6 14 CYS HB3 H 2.987 0.01 2 32 6 14 CYS N N 116.724 0.1 1 33 7 15 SER H H 9.659 0.01 1 34 7 15 SER HA H 4.226 0.01 1 35 7 15 SER HB2 H 3.953 0.01 2 36 7 15 SER HB3 H 3.858 0.01 2 37 7 15 SER N N 123.545 0.1 1 38 8 16 SER H H 7.862 0.01 1 39 8 16 SER HA H 5.137 0.01 1 40 8 16 SER HB2 H 4.005 0.01 2 41 8 16 SER HB3 H 3.816 0.01 2 42 8 16 SER N N 119.712 0.1 1 43 9 17 PRO HA H 4.559 0.01 2 44 9 17 PRO HB2 H 2.392 0.01 1 45 9 17 PRO HB3 H 2.392 0.01 1 46 9 17 PRO HG2 H 2.040 0.01 1 47 9 17 PRO HG3 H 2.040 0.01 1 48 9 17 PRO HD2 H 3.965 0.01 2 49 9 17 PRO HD3 H 3.824 0.01 2 50 10 18 GLU H H 8.244 0.01 1 51 10 18 GLU HA H 4.294 0.01 1 52 10 18 GLU HB2 H 2.201 0.01 2 53 10 18 GLU HB3 H 2.031 0.01 2 54 10 18 GLU HG2 H 2.261 0.01 1 55 10 18 GLU HG3 H 2.261 0.01 1 56 10 18 GLU N N 120.855 0.1 1 57 11 19 ASN H H 7.260 0.01 1 58 11 19 ASN HA H 4.794 0.01 1 59 11 19 ASN HB2 H 3.146 0.01 2 60 11 19 ASN HB3 H 2.872 0.01 2 61 11 19 ASN HD21 H 8.296 0.01 2 62 11 19 ASN HD22 H 7.438 0.01 2 63 11 19 ASN N N 124.776 0.1 1 64 11 19 ASN ND2 N 118.519 0.1 1 65 12 20 PRO HA H 4.704 0.01 2 66 12 20 PRO HB2 H 2.393 0.01 2 67 12 20 PRO HB3 H 1.979 0.01 2 68 12 20 PRO HG2 H 2.151 0.01 1 69 12 20 PRO HG3 H 2.151 0.01 1 70 12 20 PRO HD2 H 4.165 0.01 2 71 12 20 PRO HD3 H 3.969 0.01 2 72 13 21 CYS H H 8.931 0.01 1 73 13 21 CYS HA H 4.648 0.01 1 74 13 21 CYS HB2 H 3.222 0.01 2 75 13 21 CYS HB3 H 2.578 0.01 2 76 13 21 CYS N N 121.945 0.1 1 77 14 22 CYS H H 7.543 0.01 1 78 14 22 CYS HA H 5.056 0.01 1 79 14 22 CYS HB2 H 2.95 0.01 2 80 14 22 CYS HB3 H 2.89 0.01 2 81 14 22 CYS N N 116.46 0.1 1 82 15 23 ASP H H 8.212 0.01 1 83 15 23 ASP HA H 4.570 0.01 1 84 15 23 ASP HB2 H 3.144 0.01 2 85 15 23 ASP HB3 H 2.481 0.01 2 86 15 23 ASP N N 125.197 0.1 1 87 16 24 ALA H H 8.996 0.01 1 88 16 24 ALA HA H 4.148 0.01 1 89 16 24 ALA HB H 1.546 0.01 1 90 16 24 ALA N N 135.341 0.1 1 91 17 25 ALA H H 8.669 0.01 1 92 17 25 ALA HA H 4.361 0.01 1 93 17 25 ALA HB H 1.544 0.01 1 94 17 25 ALA N N 109.481 0.1 1 95 18 26 THR H H 7.743 0.01 1 96 18 26 THR HA H 4.445 0.01 2 97 18 26 THR HB H 4.34 0.01 2 98 18 26 THR HG2 H 1.198 0.01 1 99 18 26 THR N N 109.481 0.1 1 100 19 27 CYS H H 8.601 0.01 1 101 19 27 CYS HA H 4.642 0.01 1 102 19 27 CYS HB2 H 3.543 0.01 2 103 19 27 CYS HB3 H 3.323 0.01 2 104 19 27 CYS N N 123.176 0.1 1 105 20 28 LYS H H 7.964 0.01 1 106 20 28 LYS HA H 4.945 0.01 1 107 20 28 LYS HB2 H 1.984 0.01 2 108 20 28 LYS HB3 H 1.536 0.01 2 109 20 28 LYS HD2 H 1.675 0.01 1 110 20 28 LYS HD3 H 1.675 0.01 1 111 20 28 LYS N N 122.525 0.1 1 112 21 29 LEU H H 9.211 0.01 1 113 21 29 LEU HA H 4.285 0.01 1 114 21 29 LEU HB2 H 1.424 0.01 1 115 21 29 LEU HB3 H 1.424 0.01 1 116 21 29 LEU HG H 1.681 0.01 2 117 21 29 LEU HD1 H 0.632 0.01 2 118 21 29 LEU HD2 H 0.519 0.01 2 119 21 29 LEU N N 124.582 0.1 1 120 22 30 ARG H H 8.490 0.01 1 121 22 30 ARG HA H 4.448 0.01 1 122 22 30 ARG HB2 H 1.719 0.01 2 123 22 30 ARG HB3 H 1.627 0.01 2 124 22 30 ARG HD2 H 3.249 0.01 2 125 22 30 ARG HD3 H 3.172 0.01 2 126 22 30 ARG HE H 6.925 0.01 2 127 22 30 ARG N N 126.604 0.1 1 128 22 30 ARG NE N 125.132 0.1 1 129 23 31 PRO HA H 4.389 0.01 2 130 23 31 PRO HB2 H 2.378 0.01 2 131 23 31 PRO HB3 H 1.921 0.01 2 132 23 31 PRO HG2 H 2.091 0.01 1 133 23 31 PRO HG3 H 2.091 0.01 1 134 24 32 GLY H H 8.830 0.01 1 135 24 32 GLY HA2 H 4.295 0.01 2 136 24 32 GLY HA3 H 3.664 0.01 2 137 24 32 GLY N N 117.708 0.1 1 138 25 33 ALA H H 7.942 0.01 1 139 25 33 ALA HA H 4.540 0.01 1 140 25 33 ALA HB H 1.621 0.01 1 141 25 33 ALA N N 127.553 0.1 1 142 26 34 GLN H H 8.827 0.01 1 143 26 34 GLN HA H 4.113 0.01 1 144 26 34 GLN HB2 H 1.236 0.01 2 145 26 34 GLN HB3 H 0.574 0.01 2 146 26 34 GLN HG2 H 2.168 0.01 2 147 26 34 GLN HG3 H 2.056 0.01 2 148 26 34 GLN HE21 H 6.988 0.01 2 149 26 34 GLN HE22 H 7.717 0.01 2 150 26 34 GLN N N 122.086 0.1 1 151 26 34 GLN NE2 N 117.676 0.1 1 152 27 35 CYS H H 7.805 0.01 1 153 27 35 CYS HA H 4.555 0.01 1 154 27 35 CYS HB2 H 3.287 0.01 2 155 27 35 CYS HB3 H 3.153 0.01 2 156 27 35 CYS N N 117.409 0.1 1 157 28 36 GLY H H 9.163 0.01 1 158 28 36 GLY HA2 H 4.446 0.01 1 159 28 36 GLY HA3 H 3.574 0.01 1 160 28 36 GLY N N 109.041 0.1 1 161 29 37 GLU H H 7.844 0.01 1 162 29 37 GLU HA H 4.782 0.01 1 163 29 37 GLU HB2 H 2.311 0.01 2 164 29 37 GLU HB3 H 1.981 0.01 2 165 29 37 GLU HG2 H 2.260 0.01 2 166 29 37 GLU HG3 H 2.171 0.01 2 167 29 37 GLU N N 122.174 0.1 1 168 30 38 GLY H H 8.391 0.01 1 169 30 38 GLY HA2 H 4.915 0.01 2 170 30 38 GLY HA3 H 3.925 0.01 2 171 30 38 GLY N N 112.663 0.1 1 172 31 39 LEU H H 8.960 0.01 1 173 31 39 LEU HA H 4.354 0.01 1 174 31 39 LEU HB2 H 1.829 0.01 2 175 31 39 LEU HB3 H 1.741 0.01 2 176 31 39 LEU HG H 1.987 0.01 2 177 31 39 LEU HD1 H 1.142 0.01 1 178 31 39 LEU HD2 H 1.142 0.01 1 179 31 39 LEU N N 125.039 0.1 1 180 32 40 CYS H H 8.467 0.01 1 181 32 40 CYS HA H 5.108 0.01 1 182 32 40 CYS HB2 H 3.894 0.01 2 183 32 40 CYS HB3 H 2.509 0.01 2 184 32 40 CYS N N 119.167 0.1 1 185 33 41 CYS H H 7.365 0.01 1 186 33 41 CYS HA H 5.291 0.01 1 187 33 41 CYS HB2 H 2.994 0.01 2 188 33 41 CYS HB3 H 2.560 0.01 2 189 33 41 CYS N N 125.303 0.1 1 190 34 42 GLU H H 9.742 0.01 1 191 34 42 GLU HA H 4.682 0.01 1 192 34 42 GLU HB2 H 2.055 0.01 2 193 34 42 GLU HB3 H 1.911 0.01 2 194 34 42 GLU HG2 H 2.228 0.01 1 195 34 42 GLU HG3 H 2.228 0.01 1 196 34 42 GLU N N 132.652 0.1 1 197 35 43 GLN H H 9.453 0.01 1 198 35 43 GLN HA H 3.999 0.01 1 199 35 43 GLN HB2 H 2.399 0.01 2 200 35 43 GLN HB3 H 2.128 0.01 2 201 35 43 GLN HG2 H 2.241 0.01 1 202 35 43 GLN HG3 H 2.241 0.01 1 203 35 43 GLN N N 129.593 0.1 1 204 35 43 GLN NE2 N 116.264 0.1 1 205 36 44 CYS H H 8.492 0.01 1 206 36 44 CYS HA H 4.801 0.01 1 207 36 44 CYS HB2 H 3.763 0.01 2 208 36 44 CYS HB3 H 3.372 0.01 2 209 36 44 CYS N N 109.569 0.1 1 210 37 45 LYS H H 8.024 0.01 1 211 37 45 LYS HA H 4.703 0.01 1 212 37 45 LYS HB2 H 1.88 0.01 2 213 37 45 LYS HB3 H 1.79 0.01 2 214 37 45 LYS HG2 H 1.46 0.01 2 215 37 45 LYS HG3 H 1.54 0.01 2 216 37 45 LYS HD2 H 1.64 0.01 1 217 37 45 LYS HD3 H 1.64 0.01 1 218 37 45 LYS HE2 H 3.039 0.01 1 219 37 45 LYS HE3 H 3.039 0.01 1 220 37 45 LYS N N 125.11 0.1 1 221 38 46 PHE H H 8.276 0.01 1 222 38 46 PHE HA H 5.141 0.01 1 223 38 46 PHE HB2 H 3.050 0.01 2 224 38 46 PHE HB3 H 2.724 0.01 2 225 38 46 PHE HD1 H 7.029 0.01 1 226 38 46 PHE HD2 H 7.029 0.01 1 227 38 46 PHE HE1 H 7.261 0.01 1 228 38 46 PHE HE2 H 7.261 0.01 1 229 38 46 PHE N N 123.897 0.1 1 230 39 47 SER H H 9.044 0.01 1 231 39 47 SER HA H 4.274 0.01 1 232 39 47 SER HB2 H 3.932 0.01 2 233 39 47 SER HB3 H 3.765 0.01 2 234 39 47 SER N N 125.655 0.1 1 235 40 48 ARG H H 8.573 0.01 1 236 40 48 ARG HA H 4.101 0.01 1 237 40 48 ARG HB2 H 1.82 0.01 1 238 40 48 ARG HB3 H 1.82 0.01 1 239 40 48 ARG HG2 H 1.688 0.01 1 240 40 48 ARG HG3 H 1.688 0.01 1 241 40 48 ARG HD2 H 3.269 0.01 1 242 40 48 ARG HD3 H 3.269 0.01 1 243 40 48 ARG N N 125.848 0.1 1 244 41 49 ALA H H 8.501 0.01 1 245 41 49 ALA HA H 3.724 0.01 1 246 41 49 ALA HB H 1.298 0.01 1 247 41 49 ALA N N 127.729 0.1 1 248 42 50 GLY H H 8.619 0.01 1 249 42 50 GLY HA2 H 4.486 0.01 2 250 42 50 GLY HA3 H 3.587 0.01 2 251 42 50 GLY N N 114.579 0.1 1 252 43 51 LYS H H 7.535 0.01 1 253 43 51 LYS HA H 4.149 0.01 1 254 43 51 LYS HB2 H 1.87 0.01 1 255 43 51 LYS HB3 H 1.87 0.01 1 256 43 51 LYS HG2 H 1.529 0.01 2 257 43 51 LYS HG3 H 1.415 0.01 2 258 43 51 LYS HD2 H 1.697 0.01 1 259 43 51 LYS HD3 H 1.697 0.01 1 260 43 51 LYS HE2 H 3.117 0.01 1 261 43 51 LYS HE3 H 3.117 0.01 1 262 43 51 LYS N N 126.78 0.1 1 263 44 52 ILE H H 8.810 0.01 1 264 44 52 ILE HA H 4.167 0.01 1 265 44 52 ILE HB H 1.816 0.01 2 266 44 52 ILE HG2 H 1.093 0.01 1 267 44 52 ILE HD1 H 0.998 0.01 1 268 44 52 ILE N N 133.179 0.1 1 269 45 53 CYS H H 9.355 0.01 1 270 45 53 CYS HA H 5.160 0.01 1 271 45 53 CYS HB2 H 3.216 0.01 1 272 45 53 CYS HB3 H 3.216 0.01 1 273 45 53 CYS N N 127.5 0.1 1 274 46 54 ARG H H 7.675 0.01 1 275 46 54 ARG HA H 4.469 0.01 1 276 46 54 ARG HB2 H 2.014 0.01 1 277 46 54 ARG HB3 H 2.014 0.01 1 278 46 54 ARG HG2 H 1.72 0.01 2 279 46 54 ARG HG3 H 1.49 0.01 2 280 46 54 ARG HD2 H 3.396 0.01 1 281 46 54 ARG HD3 H 3.396 0.01 1 282 46 54 ARG N N 126.375 0.1 1 283 47 55 ILE H H 8.400 0.01 1 284 47 55 ILE HA H 4.663 0.01 1 285 47 55 ILE HB H 1.897 0.01 2 286 47 55 ILE HG12 H 1.190 0.01 1 287 47 55 ILE HG13 H 1.190 0.01 1 288 47 55 ILE HG2 H 1.031 0.01 1 289 47 55 ILE HD1 H 0.930 0.01 1 290 47 55 ILE N N 129.698 0.1 1 291 48 56 PRO HA H 4.491 0.01 1 292 48 56 PRO HB2 H 2.304 0.01 1 293 48 56 PRO HB3 H 2.304 0.01 1 294 48 56 PRO HG2 H 2.007 0.01 1 295 48 56 PRO HG3 H 2.007 0.01 1 296 48 56 PRO HD2 H 3.902 0.01 1 297 48 56 PRO HD3 H 3.902 0.01 1 298 49 57 ARG H H 8.331 0.01 1 299 49 57 ARG HA H 4.398 0.01 1 300 49 57 ARG HB2 H 1.925 0.01 2 301 49 57 ARG HB3 H 1.812 0.01 2 302 49 57 ARG HG2 H 1.729 0.01 1 303 49 57 ARG HG3 H 1.729 0.01 1 304 49 57 ARG HD2 H 3.183 0.01 2 305 49 57 ARG HD3 H 3.136 0.01 2 306 49 57 ARG N N 122.965 0.1 1 307 50 58 GLY H H 8.426 0.01 1 308 50 58 GLY HA2 H 3.958 0.01 2 309 50 58 GLY HA3 H 3.915 0.01 2 310 50 58 GLY N N 114.157 0.1 1 311 51 59 ASP H H 8.610 0.01 1 312 51 59 ASP HA H 4.659 0.01 1 313 51 59 ASP HB2 H 2.791 0.01 2 314 51 59 ASP HB3 H 2.741 0.01 2 315 51 59 ASP N N 124.951 0.1 1 316 52 60 MET H H 7.745 0.01 1 317 52 60 MET HA H 4.839 0.01 1 318 52 60 MET HB2 H 2.08 0.01 2 319 52 60 MET HB3 H 2.01 0.01 2 320 52 60 MET HG2 H 2.693 0.01 2 321 52 60 MET HG3 H 2.574 0.01 2 322 52 60 MET N N 124.354 0.1 1 323 53 61 PRO HA H 4.485 0.01 1 324 53 61 PRO HB2 H 2.249 0.01 1 325 53 61 PRO HB3 H 2.249 0.01 1 326 53 61 PRO HG2 H 1.894 0.01 1 327 53 61 PRO HG3 H 1.894 0.01 1 328 53 61 PRO HD2 H 3.797 0.01 2 329 53 61 PRO HD3 H 3.472 0.01 2 330 54 62 ASP H H 8.114 0.01 1 331 54 62 ASP HA H 4.577 0.01 1 332 54 62 ASP HB2 H 2.764 0.01 2 333 54 62 ASP HB3 H 2.54 0.01 2 334 54 62 ASP N N 122.385 0.1 1 335 55 63 ASP H H 8.388 0.01 1 336 55 63 ASP HA H 5.052 0.01 1 337 55 63 ASP HB2 H 2.953 0.01 2 338 55 63 ASP HB3 H 2.566 0.01 2 339 55 63 ASP N N 124.758 0.1 1 340 56 64 ARG H H 8.358 0.01 1 341 56 64 ARG HA H 5.332 0.01 1 342 56 64 ARG HB2 H 1.557 0.01 1 343 56 64 ARG HB3 H 1.557 0.01 1 344 56 64 ARG HG2 H 1.408 0.01 1 345 56 64 ARG HG3 H 1.408 0.01 1 346 56 64 ARG HD2 H 3.221 0.01 2 347 56 64 ARG HD3 H 3.117 0.01 2 348 56 64 ARG N N 123.703 0.1 1 349 56 64 ARG NE N 126.435 0.1 1 350 57 65 CYS H H 9.377 0.01 1 351 57 65 CYS HA H 4.862 0.01 1 352 57 65 CYS HB2 H 3.891 0.01 2 353 57 65 CYS HB3 H 2.730 0.01 2 354 57 65 CYS N N 123.897 0.1 1 355 58 66 THR H H 10.180 0.01 1 356 58 66 THR HA H 4.442 0.01 2 357 58 66 THR HB H 4.510 0.01 2 358 58 66 THR HG2 H 1.415 0.01 1 359 58 66 THR N N 117.445 0.1 1 360 59 67 GLY H H 8.546 0.01 1 361 59 67 GLY HA2 H 4.460 0.01 1 362 59 67 GLY HA3 H 4.460 0.01 1 363 59 67 GLY N N 115.88 0.1 1 364 60 68 GLN H H 7.986 0.01 1 365 60 68 GLN HA H 4.298 0.01 1 366 60 68 GLN HB2 H 2.215 0.01 2 367 60 68 GLN HB3 H 1.692 0.01 2 368 60 68 GLN N N 119.027 0.1 1 369 60 68 GLN NE2 N 115.688 0.1 1 370 61 69 SER H H 6.630 0.01 1 371 61 69 SER HA H 3.88 0.01 2 372 61 69 SER HB2 H 3.88 0.01 2 373 61 69 SER HB3 H 3.621 0.01 2 374 61 69 SER N N 114.913 0.1 1 375 62 70 ALA H H 9.027 0.01 1 376 62 70 ALA HA H 4.435 0.01 1 377 62 70 ALA HB H 1.827 0.01 1 378 62 70 ALA N N 129.628 0.1 1 379 63 71 ASP H H 8.08 0.01 1 380 63 71 ASP HA H 4.895 0.01 1 381 63 71 ASP HB2 H 2.671 0.01 2 382 63 71 ASP HB3 H 2.543 0.01 2 383 63 71 ASP N N 121.506 0.1 1 384 64 72 CYS H H 8.924 0.01 1 385 64 72 CYS HA H 5.403 0.01 1 386 64 72 CYS HB2 H 3.103 0.01 2 387 64 72 CYS HB3 H 2.802 0.01 2 388 64 72 CYS N N 125.936 0.1 1 389 65 73 PRO HA H 4.375 0.01 2 390 65 73 PRO HB2 H 2.216 0.01 2 391 65 73 PRO HB3 H 1.802 0.01 2 392 65 73 PRO HG2 H 2.130 0.01 2 393 65 73 PRO HG3 H 2.014 0.01 2 394 65 73 PRO HD2 H 3.994 0.01 2 395 65 73 PRO HD3 H 3.733 0.01 2 396 66 74 ARG H H 8.203 0.01 1 397 66 74 ARG HA H 4.362 0.01 1 398 66 74 ARG HB2 H 1.82 0.01 2 399 66 74 ARG HB3 H 1.76 0.01 2 400 66 74 ARG HG2 H 1.641 0.01 1 401 66 74 ARG HG3 H 1.641 0.01 1 402 66 74 ARG HD2 H 3.21 0.01 1 403 66 74 ARG HD3 H 3.21 0.01 1 404 66 74 ARG N N 123.58 0.1 1 405 67 75 TYR H H 8.650 0.01 1 406 67 75 TYR HA H 4.459 0.01 1 407 67 75 TYR HB2 H 3.135 0.01 2 408 67 75 TYR HB3 H 3.006 0.01 2 409 67 75 TYR HD1 H 7.163 0.01 1 410 67 75 TYR HD2 H 7.163 0.01 1 411 67 75 TYR HE1 H 6.863 0.01 1 412 67 75 TYR HE2 H 6.863 0.01 1 413 67 75 TYR N N 128.872 0.1 1 414 68 76 HIS H H 7.655 0.01 1 415 68 76 HIS HA H 4.365 0.01 1 416 68 76 HIS HB2 H 3.229 0.01 2 417 68 76 HIS HB3 H 3.09 0.01 2 418 68 76 HIS N N 129.258 0.1 1 stop_ loop_ _Atom_shift_assign_ID_ambiguity 9 8 stop_ save_