data_20049 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; NMR solution structure of Mu-KIIIA[C1A,C9A] ; _BMRB_accession_number 20049 _BMRB_flat_file_name bmr20049.str _Entry_type new _Submission_date 2008-09-28 _Accession_date 2008-09-28 _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 Khoo Keith K. . 2 Feng Zhiping . . 3 Norton Raymond S. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 88 "13C chemical shifts" 15 "15N chemical shifts" 12 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2010-06-03 update BMRB 'edit assembly name' 2009-05-18 update BMRB 'complete entry citation' 2009-04-04 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 20048 'Native toxin' stop_ save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Structure of the Analgesic mu-Conotoxin KIIIA and Effects on the Structure and Function of Disulfide Deletion' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 19170536 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Khoo Keith K. . 2 Feng Zhi-Ping . . 3 Smith Brian J. . 4 Zhang Min-Min . . 5 Yoshikami Doju . . 6 Olivera Baldomero M. . 7 Bulaj Grzegorz . . 8 Norton Raymond S. . stop_ _Journal_abbreviation Biochemistry _Journal_name_full Biochemistry _Journal_volume 48 _Journal_issue 6 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 1210 _Page_last 1219 _Year 2009 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name Mu-KIIIA[C1A,C9A] _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label KIIIA[Ala1,9] $KIIIA[Ala1,9] stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_KIIIA[Ala1,9] _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common KIIIA[Ala1,9] _Molecular_mass 1831.085 _Mol_thiol_state 'all disulfide bound' _Details ; Disulfide pairing Cys2-Cys15 , Cys4-Cys16 ; ############################## # Polymer residue sequence # ############################## _Residue_count 16 _Mol_residue_sequence ACNCSSKWARDHSRCC loop_ _Residue_seq_code _Residue_label 1 ALA 2 CYS 3 ASN 4 CYS 5 SER 6 SER 7 LYS 8 TRP 9 ALA 10 ARG 11 ASP 12 HIS 13 SER 14 ARG 15 CYS 16 CYS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ######################################## # Molecular bond linkage definitions # ######################################## save_crosslink_bonds _Saveframe_category crosslink_bonds loop_ _Bond_order _Bond_type _Atom_one_mol_system_component_name _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_atom_name _Atom_two_mol_system_component_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_atom_name 1 disulfide KIIIA[Ala1,9] 2 CYS SG KIIIA[Ala1,9] 15 CYS SG 1 disulfide KIIIA[Ala1,9] 4 CYS SG KIIIA[Ala1,9] 16 CYS SG stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $KIIIA[Ala1,9] Conuskinoshitai 376876 Eukaryota Conidae Conus kinoshitai 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 $KIIIA[Ala1,9] 'recombinant technology' . . . . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_KIIIA[Ala1,9]_H2O _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $KIIIA[Ala1,9] 3.7 mM 'natural abundance' stop_ save_ save_KIIIA[Ala1,9]_D2O _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $KIIIA[Ala1,9] 3.7 mM 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version 1.3 loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task collection processing stop_ _Details . save_ save_XEASY _Saveframe_category software _Name XEASY _Version 1.3.13 loop_ _Vendor _Address _Electronic_address 'Bartels et al.' . . stop_ loop_ _Task 'peak picking' 'chemical shift assignment' stop_ _Details . save_ save_CYANA _Saveframe_category software _Name CYANA _Version . loop_ _Vendor _Address _Electronic_address 'Guntert, Mumenthaler and Wuthrich' . . stop_ loop_ _Task 'structure solution' stop_ _Details . save_ save_X-PLOR_NIH _Saveframe_category software _Name X-PLOR_NIH _Version . loop_ _Vendor _Address _Electronic_address 'Schwieters, Kuszewski, Tjandra and Clore' . . stop_ loop_ _Task refinement 'geometry optimization' 'structure solution' stop_ _Details . save_ save_ProcheckNMR _Saveframe_category software _Name ProcheckNMR _Version . loop_ _Vendor _Address _Electronic_address 'Laskowski and MacArthur' . . stop_ loop_ _Task 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model DRX _Field_strength 600 _Details . save_ save_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_1H-13C_HSQC_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HSQC' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_1H-1H_TOCSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_DQF-COSY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D DQF-COSY' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_1H-1H_NOESY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_1H-1H_TOCSY_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_1H-1H_NOESY_7 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $KIIIA[Ala1,9]_H2O save_ save_2D_1H-1H_TOCSY_8 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $KIIIA[Ala1,9]_D2O save_ save_2D_1H-1H_NOESY_9 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $KIIIA[Ala1,9]_D2O save_ save_2D_1H-1H_NOESY_10 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $KIIIA[Ala1,9]_H2O save_ ####################### # Sample conditions # ####################### save_pH_5.3,_278_K _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 278 . K pH 5.3 . pH stop_ save_ save_pH_5.3,_283_K _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 283 . K pH 5.3 . pH stop_ save_ save_pH_5.3,_293_K _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 293 . K pH 5.3 . pH stop_ save_ save_pH_3.2,_278_K _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 278 . K pH 3.2 . pH stop_ save_ save_pH_4.2,_278_K _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 278 . K pH 4.2 . pH stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chemical_shift_reference_1 _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 water H 1 protons ppm 5.004 internal direct . . . 1.000000000 water C 13 protons ppm 5.004 internal indirect . . . 0.251449530 water N 15 protons ppm 5.004 internal 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_assigned_chem_shift_list_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-15N HSQC' '2D 1H-13C HSQC' '2D 1H-1H TOCSY' '2D 1H-1H NOESY' stop_ loop_ _Sample_label $KIIIA[Ala1,9]_H2O stop_ _Sample_conditions_label $pH_5.3,_278_K _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name KIIIA[Ala1,9] _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 1 ALA HA H 4.06 0.02 1 2 1 1 ALA HB H 1.50 0.02 1 3 1 1 ALA CA C 51.8 0.5 1 4 2 2 CYS H H 8.93 0.02 1 5 2 2 CYS HA H 4.58 0.02 1 6 2 2 CYS HB2 H 3.09 0.02 2 7 2 2 CYS HB3 H 2.99 0.02 2 8 2 2 CYS CA C 56.3 0.5 1 9 3 3 ASN H H 8.92 0.02 1 10 3 3 ASN HA H 4.81 0.02 1 11 3 3 ASN HB2 H 3.01 0.02 2 12 3 3 ASN HB3 H 2.79 0.02 2 13 3 3 ASN HD21 H 7.86 0.02 2 14 3 3 ASN HD22 H 7.17 0.02 2 15 3 3 ASN CA C 52.9 0.5 1 16 4 4 CYS H H 8.91 0.02 1 17 4 4 CYS HA H 4.96 0.02 1 18 4 4 CYS HB2 H 3.44 0.02 2 19 4 4 CYS HB3 H 2.63 0.02 2 20 5 5 SER H H 8.23 0.02 1 21 5 5 SER HA H 4.31 0.02 1 22 5 5 SER HB2 H 4.09 0.02 2 23 5 5 SER HB3 H 4.00 0.02 2 24 5 5 SER HG H 6.15 0.02 1 25 5 5 SER CA C 60.7 0.5 1 26 5 5 SER N N 116.5 0.5 5 27 6 6 SER H H 7.72 0.02 1 28 6 6 SER HA H 4.83 0.02 1 29 6 6 SER HB2 H 4.33 0.02 2 30 6 6 SER HB3 H 4.02 0.02 2 31 6 6 SER HG H 6.31 0.02 1 32 6 6 SER CA C 56.2 0.5 1 33 6 6 SER N N 115.4 0.5 1 34 7 7 LYS H H 9.13 0.02 1 35 7 7 LYS HA H 3.90 0.02 1 36 7 7 LYS HB2 H 1.89 0.02 2 37 7 7 LYS HB3 H 1.89 0.02 2 38 7 7 LYS HD2 H 1.78 0.02 2 39 7 7 LYS HD3 H 1.65 0.02 2 40 7 7 LYS HE2 H 3.05 0.02 2 41 7 7 LYS HE3 H 3.05 0.02 2 42 7 7 LYS HG2 H 1.45 0.02 2 43 7 7 LYS HG3 H 1.45 0.02 2 44 7 7 LYS HZ H 7.81 0.02 1 45 7 7 LYS CA C 59.7 0.02 1 46 7 7 LYS N N 124.9 0.02 1 47 8 8 TRP H H 8.80 0.02 1 48 8 8 TRP HA H 4.34 0.02 1 49 8 8 TRP HB2 H 3.44 0.02 2 50 8 8 TRP HB3 H 3.15 0.02 2 51 8 8 TRP HD1 H 7.14 0.02 1 52 8 8 TRP HE1 H 10.04 0.02 1 53 8 8 TRP HE3 H 7.48 0.02 1 54 8 8 TRP CA C 61.9 0.5 1 55 8 8 TRP N N 117.7 0.5 1 56 9 9 ALA H H 8.30 0.02 1 57 9 9 ALA HA H 4.14 0.02 1 58 9 9 ALA HB H 1.68 0.02 1 59 9 9 ALA CA C 55.5 0.5 1 60 9 9 ALA N N 122.5 0.5 1 61 10 10 ARG H H 8.26 0.02 1 62 10 10 ARG HA H 3.99 0.02 1 63 10 10 ARG HB2 H 2.00 0.02 2 64 10 10 ARG HB3 H 1.84 0.02 2 65 10 10 ARG HD2 H 3.43 0.02 2 66 10 10 ARG HD3 H 3.20 0.02 2 67 10 10 ARG HE H 7.47 0.02 1 68 10 10 ARG HG2 H 1.93 0.02 2 69 10 10 ARG HG3 H 1.62 0.02 2 70 10 10 ARG CA C 60.2 0.5 1 71 10 10 ARG N N 118.0 0.5 1 72 11 11 ASP H H 8.24 0.02 1 73 11 11 ASP HA H 4.33 0.02 1 74 11 11 ASP HB2 H 2.34 0.02 2 75 11 11 ASP HB3 H 2.00 0.02 2 76 11 11 ASP CA C 55.5 0.5 1 77 11 11 ASP N N 116.5 0.5 5 78 12 12 HIS H H 7.64 0.02 1 79 12 12 HIS HA H 4.49 0.02 1 80 12 12 HIS HB2 H 3.03 0.02 2 81 12 12 HIS HB3 H 2.03 0.02 2 82 12 12 HIS HD2 H 6.75 0.02 1 83 12 12 HIS HE1 H 7.76 0.02 1 84 12 12 HIS CA C 57.2 0.5 1 85 12 12 HIS N N 114.2 0.5 1 86 13 13 SER H H 8.51 0.02 1 87 13 13 SER HA H 4.75 0.02 1 88 13 13 SER HB2 H 4.11 0.02 2 89 13 13 SER HB3 H 4.11 0.02 2 90 13 13 SER HG H 6.01 0.02 1 91 13 13 SER CA C 57.9 0.5 1 92 13 13 SER N N 117.1 0.5 1 93 14 14 ARG H H 9.32 0.02 1 94 14 14 ARG HA H 4.09 0.02 1 95 14 14 ARG HB2 H 1.88 0.02 2 96 14 14 ARG HB3 H 1.88 0.02 2 97 14 14 ARG HD2 H 3.21 0.02 2 98 14 14 ARG HD3 H 3.21 0.02 2 99 14 14 ARG HE H 7.26 0.02 1 100 14 14 ARG HG2 H 1.69 0.02 2 101 14 14 ARG HG3 H 1.69 0.02 2 102 14 14 ARG CA C 58.6 0.5 1 103 14 14 ARG N N 130.1 0.5 1 104 15 15 CYS H H 8.71 0.02 1 105 15 15 CYS HA H 4.59 0.02 1 106 15 15 CYS HB2 H 3.45 0.02 2 107 15 15 CYS HB3 H 3.19 0.02 2 108 15 15 CYS CA C 57.2 0.5 1 109 15 15 CYS N N 115.0 0.5 1 110 16 16 CYS H H 7.76 0.02 1 111 16 16 CYS HA H 4.55 0.02 1 112 16 16 CYS HB2 H 3.27 0.02 2 113 16 16 CYS HB3 H 3.04 0.02 2 114 16 16 CYS CA C 55.9 0.5 1 115 16 16 CYS N N 121.8 0.5 1 stop_ save_ save_constraint_statistics _Saveframe_category constraint_statistics _Text_data_format . _Text_data . _NOE_interproton_distance_evaluation . _NOE_pseudoatom_corrections . _Details . save_