data_4246 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 3D Structure of the M8L Mutant of Squash Trypsin Inhibitor CMTI-I, NMR, 6 Structures ; _BMRB_accession_number 4246 _BMRB_flat_file_name bmr4246.str _Entry_type original _Submission_date 1998-10-15 _Accession_date 1998-10-15 _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 Zhukov Igor Yu . 2 Bolewska Krystina . . 3 Bierzynski Andrzej . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 166 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2000-05-02 original author . stop_ _Original_release_date 2000-05-02 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full ; Zhukov, I., Bolewska, K., and Bierzynski, A., "Conservative Mutation Met8-->Leu Affects the Folding Process and Structural Stability of Squash Trypsin Inhibitor CMTI-I," Protein Sci. 9, 273-279 (2000). ; _Citation_title ; Conservative Mutation Met8-->Leu Affects the Folding Process and Structural Stability of Squash Trypsin Inhibitor CMTI-I ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code 20178923 _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Zhukov Igor Yu . 2 Bolewska Krystina . . 3 Bierzynski Andrzej . . stop_ _Journal_abbreviation 'Protein Sci.' _Journal_name_full 'Protein Science' _Journal_volume 9 _Journal_issue . _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 273 _Page_last 279 _Year 2000 _Details . loop_ _Keyword 'serine protease inhibitor' 'trypsin inhibitor' stop_ save_ ####################################### # Cited references within the entry # ####################################### save_citation_one _Saveframe_category citation _Citation_full ; Wishart, D. S., Bigam, C. G., Yao, J., Abildgaard, F., Dyson, H. J., Oldfield, E., Markley, J. L., and Sykes, B. D. J. Biomol. NMR 6, 135-140 (1995). ; _Citation_title '1H, 13C and 15N chemical shift referencing in biomolecular NMR.' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 8589602 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Wishart 'D S' S. . 2 Bigam 'C G' G. . 3 Yao J . . 4 Abildgaard F . . 5 Dyson 'H J' J. . 6 Oldfield E . . 7 Markley 'J L' L. . 8 Sykes 'B D' D. . stop_ _Journal_abbreviation 'J. Biomol. NMR' _Journal_name_full 'Journal of biomolecular NMR' _Journal_volume 6 _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 140 _Year 1995 _Details ; A considerable degree of variability exists in the way that 1H, 13C and 15N chemical shifts are reported and referenced for biomolecules. In this article we explore some of the reasons for this situation and propose guidelines for future chemical shift referencing and for conversion from many common 1H, 13C and 15N chemical shift standards, now used in biomolecular NMR, to those proposed here. ; save_ save_citation_two _Saveframe_category citation _Citation_full ; Bartels, C., Xia, T., Guntert, P., Billeter, M. & Wuthrich,K. The program Xeasy for computer-supported NMR spectral analysis. J. Biomol. NMR 5, 1-10 (1995). ; _Citation_title . _Citation_status . _Citation_type . _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? _Journal_abbreviation . _Journal_name_full . _Journal_volume . _Journal_issue . _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 . _Page_last . _Year . _Details . save_ ################################## # Molecular system description # ################################## save_system_CMTI-I(M8L) _Saveframe_category molecular_system _Mol_system_name CMTI-I(M8L) _Abbreviation_common CMTI-I(M8L) _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label CMTI-I(M8L) $CMTI-I(M8L) stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state monomer _System_paramagnetic no _System_thiol_state 'not reported' loop_ _Biological_function 'trypsin inhibitor' stop_ _Database_query_date . _Details 'Residue Met8 has been replaced by Leu' save_ ######################## # Monomeric polymers # ######################## save_CMTI-I(M8L) _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common CMTI-I(M8L) _Name_variant M8L _Abbreviation_common CMTI-I _Molecular_mass . _Mol_thiol_state 'not reported' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 29 _Mol_residue_sequence ; RVCPRILLECKKDSDCLAEC VCLEHGYCG ; loop_ _Residue_seq_code _Residue_label 1 ARG 2 VAL 3 CYS 4 PRO 5 ARG 6 ILE 7 LEU 8 LEU 9 GLU 10 CYS 11 LYS 12 LYS 13 ASP 14 SER 15 ASP 16 CYS 17 LEU 18 ALA 19 GLU 20 CYS 21 VAL 22 CYS 23 LEU 24 GLU 25 HIS 26 GLY 27 TYR 28 CYS 29 GLY stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2008-08-19 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 PDB 1LU0 'Atomic Resolution Structure Of Squash Trypsin Inhibitor: Unexpected Metal Coordination' 100.00 29 100.00 100.00 9.62e-08 PDB 2V1V '3d Structure Of The M8l Mutant Of Squash Trypsin Inhibitor Cmti-I' 100.00 29 100.00 100.00 9.62e-08 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Organ $CMTI-I(M8L) squash 3661 Eukaryota Viridiplantae Cucurbita maxima seed 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 $CMTI-I(M8L) 'recombinant technology' 'E. coli' Escherichia coli BL21(DE3) plasmid pLys_S stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_one _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_max_value _Isotopic_labeling $CMTI-I(M8L) . mM 10.0 . NaCl 50 mM . . stop_ save_ ############################ # Computer software used # ############################ save_software_one _Saveframe_category software _Name XEASY _Version 3.21 loop_ _Task 'assignment and integration of two-dimensional NMR spectra' stop_ _Details . _Citation_label $citation_two save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_one _Saveframe_category NMR_spectrometer _Manufacturer Varian _Model UNITYplus _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_NOESY_1 _Saveframe_category NMR_applied_experiment _Experiment_name NOESY _Sample_label $sample_one save_ save_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name TOCSY _Sample_label $sample_one save_ save_DQF-COSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name DQF-COSY _Sample_label $sample_one save_ save_NMR_spec_expt__0_1 _Saveframe_category NMR_applied_experiment _Experiment_name NOESY _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_2 _Saveframe_category NMR_applied_experiment _Experiment_name TOCSY _BMRB_pulse_sequence_accession_number . _Details . save_ save_NMR_spec_expt__0_3 _Saveframe_category NMR_applied_experiment _Experiment_name DQF-COSY _BMRB_pulse_sequence_accession_number . _Details . save_ ####################### # Sample conditions # ####################### save_sample_conditions_one _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 5.6 0.2 n/a pressure 1 . atm temperature 303 0.2 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference_one _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 TSP H 1 'methyl protons' ppm 0.00 internal direct spherical internal parallel_to_Bo 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_assignment_one _Saveframe_category assigned_chemical_shifts _Details . loop_ _Sample_label $sample_one stop_ _Sample_conditions_label $sample_conditions_one _Chem_shift_reference_set_label $chemical_shift_reference_one _Mol_system_component_name CMTI-I(M8L) _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 ARG HA H 4.12 . 1 2 . 1 ARG HB2 H 1.89 . 1 3 . 1 ARG HB3 H 1.89 . 1 4 . 1 ARG HG2 H 1.58 . 2 5 . 1 ARG HG3 H 1.42 . 2 6 . 1 ARG HD2 H 3.30 . 2 7 . 1 ARG HD3 H 3.24 . 2 8 . 1 ARG HE H 7.37 . 1 9 . 2 VAL H H 9.14 . 1 10 . 2 VAL HA H 4.10 . 1 11 . 2 VAL HB H 2.05 . 1 12 . 2 VAL HG1 H 0.94 . 2 13 . 2 VAL HG2 H 0.89 . 2 14 . 3 CYS H H 8.55 . 1 15 . 3 CYS HA H 5.23 . 1 16 . 3 CYS HB2 H 2.80 . 2 17 . 3 CYS HB3 H 2.72 . 2 18 . 4 PRO HA H 4.46 . 1 19 . 4 PRO HB2 H 2.33 . 2 20 . 4 PRO HB3 H 1.84 . 2 21 . 4 PRO HG2 H 1.99 . 1 22 . 4 PRO HG3 H 1.99 . 1 23 . 4 PRO HD2 H 3.85 . 2 24 . 4 PRO HD3 H 3.63 . 2 25 . 5 ARG H H 8.49 . 1 26 . 5 ARG HA H 4.40 . 1 27 . 5 ARG HB2 H 1.94 . 1 28 . 5 ARG HB3 H 1.94 . 1 29 . 5 ARG HG2 H 1.74 . 1 30 . 5 ARG HG3 H 1.74 . 1 31 . 5 ARG HD2 H 3.25 . 1 32 . 5 ARG HD3 H 3.25 . 1 33 . 5 ARG HE H 7.21 . 1 34 . 6 ILE H H 7.48 . 1 35 . 6 ILE HA H 4.27 . 1 36 . 6 ILE HB H 1.73 . 1 37 . 6 ILE HG2 H 1.03 . 1 38 . 6 ILE HG12 H 1.37 . 1 39 . 6 ILE HG13 H 1.37 . 1 40 . 6 ILE HD1 H 0.86 . 1 41 . 7 LEU H H 8.62 . 1 42 . 7 LEU HA H 4.43 . 1 43 . 7 LEU HB2 H 1.73 . 1 44 . 7 LEU HB3 H 1.73 . 1 45 . 7 LEU HG H 1.46 . 1 46 . 7 LEU HD1 H 0.76 . 2 47 . 7 LEU HD2 H 0.69 . 2 48 . 8 LEU H H 8.89 . 1 49 . 8 LEU HA H 4.65 . 1 50 . 8 LEU HB2 H 1.62 . 1 51 . 8 LEU HB3 H 1.62 . 1 52 . 8 LEU HG H 1.32 . 1 53 . 8 LEU HD1 H 1.02 . 1 54 . 8 LEU HD2 H 1.02 . 1 55 . 9 GLU H H 8.63 . 1 56 . 9 GLU HA H 4.37 . 1 57 . 9 GLU HB2 H 1.87 . 1 58 . 9 GLU HB3 H 1.87 . 1 59 . 9 GLU HG2 H 2.19 . 2 60 . 9 GLU HG3 H 2.03 . 2 61 . 10 CYS H H 8.13 . 1 62 . 10 CYS HA H 4.88 . 1 63 . 10 CYS HB2 H 3.18 . 2 64 . 10 CYS HB3 H 3.03 . 2 65 . 11 LYS H H 9.35 . 1 66 . 11 LYS HA H 4.42 . 1 67 . 11 LYS HB2 H 1.86 . 2 68 . 11 LYS HB3 H 1.75 . 2 69 . 11 LYS HG2 H 1.42 . 1 70 . 11 LYS HG3 H 1.42 . 1 71 . 11 LYS HD2 H 1.65 . 1 72 . 11 LYS HD3 H 1.65 . 1 73 . 11 LYS HE2 H 3.00 . 1 74 . 11 LYS HE3 H 3.00 . 1 75 . 12 LYS H H 8.09 . 1 76 . 12 LYS HB2 H 1.98 . 2 77 . 12 LYS HB3 H 1.82 . 2 78 . 12 LYS HG2 H 1.39 . 2 79 . 12 LYS HG3 H 1.24 . 2 80 . 12 LYS HD2 H 1.64 . 1 81 . 12 LYS HD3 H 1.64 . 1 82 . 12 LYS HE2 H 2.99 . 1 83 . 12 LYS HE3 H 2.99 . 1 84 . 12 LYS HZ H 8.46 . 1 85 . 13 ASP H H 9.04 . 1 86 . 13 ASP HA H 4.13 . 1 87 . 13 ASP HB2 H 2.72 . 2 88 . 13 ASP HB3 H 2.80 . 2 89 . 14 SER H H 8.09 . 1 90 . 14 SER HA H 4.31 . 1 91 . 14 SER HB2 H 4.14 . 2 92 . 14 SER HB3 H 3.86 . 2 93 . 15 ASP H H 7.76 . 1 94 . 15 ASP HA H 4.60 . 1 95 . 15 ASP HB2 H 3.00 . 1 96 . 15 ASP HB3 H 3.00 . 1 97 . 16 CYS H H 7.76 . 1 98 . 16 CYS HA H 4.63 . 1 99 . 16 CYS HB2 H 2.89 . 1 100 . 16 CYS HB3 H 2.89 . 1 101 . 17 LEU H H 7.60 . 1 102 . 17 LEU HA H 4.34 . 1 103 . 17 LEU HB2 H 1.65 . 1 104 . 17 LEU HB3 H 1.65 . 1 105 . 17 LEU HG H 1.54 . 1 106 . 17 LEU HD1 H 0.89 . 1 107 . 17 LEU HD2 H 0.89 . 1 108 . 18 ALA H H 8.57 . 1 109 . 18 ALA HA H 4.01 . 1 110 . 18 ALA HB H 1.50 . 1 111 . 19 GLU H H 8.50 . 1 112 . 19 GLU HA H 4.51 . 1 113 . 19 GLU HB2 H 2.18 . 1 114 . 19 GLU HB3 H 2.18 . 1 115 . 19 GLU HG2 H 2.32 . 1 116 . 19 GLU HG3 H 2.32 . 1 117 . 20 CYS H H 8.45 . 1 118 . 20 CYS HA H 4.59 . 1 119 . 20 CYS HB2 H 3.31 . 1 120 . 20 CYS HB3 H 3.31 . 1 121 . 21 VAL H H 9.36 . 1 122 . 21 VAL HA H 4.41 . 1 123 . 21 VAL HB H 2.20 . 1 124 . 21 VAL HG1 H 0.84 . 2 125 . 21 VAL HG2 H 0.75 . 2 126 . 22 CYS H H 9.26 . 1 127 . 22 CYS HA H 4.86 . 1 128 . 22 CYS HB2 H 2.80 . 2 129 . 22 CYS HB3 H 2.44 . 2 130 . 23 LEU H H 8.14 . 1 131 . 23 LEU HA H 4.43 . 1 132 . 23 LEU HB2 H 2.01 . 2 133 . 23 LEU HB3 H 1.66 . 2 134 . 23 LEU HG H 1.85 . 1 135 . 23 LEU HD1 H 0.98 . 2 136 . 23 LEU HD2 H 0.84 . 2 137 . 24 GLU H H 8.83 . 1 138 . 24 GLU HA H 4.02 . 1 139 . 24 GLU HB2 H 2.08 . 1 140 . 24 GLU HB3 H 2.08 . 1 141 . 24 GLU HG2 H 2.23 . 1 142 . 24 GLU HG3 H 2.23 . 1 143 . 25 HIS H H 8.01 . 1 144 . 25 HIS HA H 4.78 . 1 145 . 25 HIS HB2 H 3.38 . 2 146 . 25 HIS HB3 H 3.26 . 2 147 . 25 HIS HD2 H 7.12 . 1 148 . 25 HIS HE1 H 8.02 . 1 149 . 26 GLY H H 8.26 . 1 150 . 26 GLY HA2 H 4.03 . 2 151 . 26 GLY HA3 H 3.69 . 2 152 . 27 TYR H H 6.79 . 1 153 . 27 TYR HA H 5.30 . 1 154 . 27 TYR HB2 H 3.05 . 2 155 . 27 TYR HB3 H 2.46 . 2 156 . 27 TYR HD1 H 6.77 . 1 157 . 27 TYR HD2 H 6.77 . 1 158 . 27 TYR HE1 H 6.77 . 1 159 . 27 TYR HE2 H 6.77 . 1 160 . 28 CYS H H 8.52 . 1 161 . 28 CYS HA H 5.32 . 1 162 . 28 CYS HB2 H 3.20 . 2 163 . 28 CYS HB3 H 2.86 . 2 164 . 29 GLY H H 9.75 . 1 165 . 29 GLY HA2 H 4.18 . 2 166 . 29 GLY HA3 H 3.87 . 2 stop_ save_