data_20033 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Solution structure of dicarba-ImI: an alpha-conotoxin with a non-reducible cystine analogue ; _BMRB_accession_number 20033 _BMRB_flat_file_name bmr20033.str _Entry_type original _Submission_date 2008-08-13 _Accession_date 2008-08-13 _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 MacRaild Christopher A. . 2 Illesinghe Jayamini . . 3 'van Lierop' Bianca . . 4 Robinson Andrea J. . 5 Norton Raymond S. . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 structure_coordinate_set 1 coupling_constants 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 65 "13C chemical shifts" 34 "15N chemical shifts" 10 "coupling constants" 13 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2009-03-18 original author . stop_ _Original_release_date 2009-03-18 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title 'Structure and activity of (2,8)-dicarba-(3,12)-cystino alpha-ImI, an alpha-conotoxin containing a nonreducible cystine analogue' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 19125616 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 MacRaild Christopher A. . 2 Illesinghe Jayamini . . 3 'van Lierop' Bianca . . 4 Townsend Amanda L. . 5 Chebib Mary . . 6 Livett Bruce G. . 7 Robinson Andrea J. . 8 Norton Raymond S. . stop_ _Journal_abbreviation 'J. Med. Chem.' _Journal_volume 52 _Journal_issue 3 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 755 _Page_last 762 _Year 2009 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name dicarba-ImI _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label dicarba-ImI $dicarba-ImI stop_ _System_molecular_weight . _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . loop_ _Biological_function 'nAChR antagonist' Toxin stop_ _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_dicarba-ImI _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common dicarba-ImI _Molecular_mass . _Mol_thiol_state 'all disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 13 _Mol_residue_sequence GXCSDPRXAWRCX loop_ _Residue_seq_code _Residue_label 1 GLY 2 ABA 3 CYS 4 SER 5 ASP 6 PRO 7 ARG 8 ABA 9 ALA 10 TRP 11 ARG 12 CYS 13 NH2 stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ###################### # Polymer residues # ###################### save_chem_comp_ABA _Saveframe_category polymer_residue _Mol_type 'L-PEPTIDE LINKING' _Name_common 'ALPHA-AMINOBUTYRIC ACID' _BMRB_code ABA _PDB_code ABA _Standard_residue_derivative . _Molecular_mass 103.120 _Mol_paramagnetic . _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? CA CA C . 0 . ? C C C . 0 . ? O O O . 0 . ? CB CB C . 0 . ? CG CG C . 0 . ? OXT OXT O . 0 . ? H H H . 0 . ? HN2 HN2 H . 0 . ? HA HA H . 0 . ? HB3 HB3 H . 0 . ? HB2 HB2 H . 0 . ? HG1 HG1 H . 0 . ? HG3 HG3 H . 0 . ? HG2 HG2 H . 0 . ? HXT HXT H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING N CA ? ? SING N H ? ? SING N HN2 ? ? SING CA C ? ? SING CA CB ? ? SING CA HA ? ? DOUB C O ? ? SING C OXT ? ? SING CB CG ? ? SING CB HB3 ? ? SING CB HB2 ? ? SING CG HG1 ? ? SING CG HG3 ? ? SING CG HG2 ? ? SING OXT HXT ? ? stop_ save_ save_chem_comp_NH2 _Saveframe_category polymer_residue _Mol_type NON-POLYMER _Name_common 'AMINO GROUP' _BMRB_code NH2 _PDB_code NH2 _Standard_residue_derivative . _Molecular_mass 16.023 _Mol_paramagnetic . _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons N N N . 0 . ? HN1 HN1 H . 0 . ? HN2 HN2 H . 0 . ? stop_ loop_ _Bond_order _Bond_atom_one_atom_name _Bond_atom_two_atom_name _PDB_bond_atom_one_atom_name _PDB_bond_atom_two_atom_name SING N HN1 ? ? SING N HN2 ? ? stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $dicarba-ImI 'Conus imperialis' 35631 Eukaryota Metazoa Conus imperialis 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 $dicarba-ImI 'chemical synthesis' . . . . . 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 $dicarba-ImI 10 mM 'natural abundance' H2O 95 % 'natural abundance' D2O 5 % 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $dicarba-ImI 10 mM 'natural abundance' D2O 100 % 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ 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 'structure solution' stop_ _Details . save_ save_XEASY _Saveframe_category software _Name XEASY _Version . loop_ _Vendor _Address _Electronic_address 'Bartels et al.' . . stop_ loop_ _Task 'chemical shift assignment' 'chemical shift calculation' 'peak picking' stop_ _Details . save_ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version . loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task collection processing 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 800 _Details . save_ save_spectrometer_3 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 500 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-1H_TOCSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $sample_1 save_ save_2D_1H-1H_NOESY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $sample_1 save_ save_2D_DQF-COSY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D DQF-COSY' _Sample_label $sample_1 save_ save_2D_1H-1H_NOESY_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $sample_2 save_ save_2D_1H-1H_eCOSY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H eCOSY' _Sample_label $sample_2 save_ save_2D_1H-15N_HSQC_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_1 save_ save_2D_1H-13C_HSQC_7 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HSQC' _Sample_label $sample_2 save_ save_2D_1H-13C_HMQC-TOCSY_8 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HMQC-TOCSY' _Sample_label $sample_2 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 5.9 . pH temperature 288 . K 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 DSS C 13 'methyl protons' ppm 0 na indirect . . . 0.251449530 DSS H 1 'methyl protons' ppm 0 internal direct . . . 1.0 DSS N 15 'methyl protons' ppm 0 na 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_ _Software_label $XEASY stop_ loop_ _Experiment_label '2D 1H-1H TOCSY' '2D 1H-1H NOESY' '2D 1H-15N HSQC' '2D 1H-13C HSQC' '2D 1H-13C HMQC-TOCSY' stop_ loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name dicarba-ImI _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 2 2 ABA HA H 4.693 0.002 1 2 2 2 ABA HB2 H 2.393 0.002 2 3 2 2 ABA HB3 H 2.718 0.002 2 4 2 2 ABA HG H 5.544 0.001 1 5 2 2 ABA CA C 56.158 0.020 1 6 2 2 ABA CB C 31.877 0.020 1 7 2 2 ABA CG C 75.928 0.020 1 8 3 3 CYS H H 8.077 0.002 1 9 3 3 CYS HA H 4.259 0.002 1 10 3 3 CYS HB2 H 2.904 0.002 2 11 3 3 CYS HB3 H 3.355 0.002 2 12 3 3 CYS CA C 56.772 0.020 1 13 3 3 CYS CB C 40.382 0.020 1 14 3 3 CYS N N 114.041 0.010 1 15 4 4 SER H H 8.502 0.001 1 16 4 4 SER HA H 4.352 0.002 1 17 4 4 SER HB2 H 3.946 0.003 2 18 4 4 SER HB3 H 3.946 0.003 2 19 4 4 SER CA C 58.846 0.020 1 20 4 4 SER CB C 63.020 0.020 1 21 4 4 SER N N 114.237 0.010 1 22 5 5 ASP H H 8.050 0.002 1 23 5 5 ASP HA H 5.074 0.002 1 24 5 5 ASP HB2 H 2.645 0.002 2 25 5 5 ASP HB3 H 3.094 0.002 2 26 5 5 ASP CA C 50.883 0.020 1 27 5 5 ASP CB C 43.122 0.020 1 28 6 6 PRO HA H 4.353 0.001 1 29 6 6 PRO HB2 H 1.966 0.002 2 30 6 6 PRO HB3 H 2.413 0.003 2 31 6 6 PRO HD2 H 3.938 0.002 2 32 6 6 PRO HD3 H 3.938 0.002 2 33 6 6 PRO HG2 H 2.055 0.005 2 34 6 6 PRO HG3 H 2.055 0.005 2 35 6 6 PRO CA C 64.929 0.020 1 36 6 6 PRO CB C 32.517 0.020 1 37 6 6 PRO CD C 51.420 0.020 1 38 6 6 PRO CG C 27.415 0.020 1 39 7 7 ARG H H 8.511 0.001 1 40 7 7 ARG HA H 4.241 0.002 1 41 7 7 ARG HB2 H 1.806 0.002 2 42 7 7 ARG HB3 H 1.899 0.033 2 43 7 7 ARG HD2 H 3.179 0.001 2 44 7 7 ARG HD3 H 3.251 0.002 2 45 7 7 ARG HE H 7.570 0.002 1 46 7 7 ARG HG2 H 1.799 0.002 2 47 7 7 ARG HG3 H 1.911 0.002 2 48 7 7 ARG CB C 27.304 0.020 1 49 7 7 ARG CD C 43.263 0.020 1 50 7 7 ARG CG C 30.316 0.020 1 51 7 7 ARG N N 116.111 0.010 1 52 7 7 ARG NE N 84.390 0.010 1 53 8 8 ABA H H 7.891 0.001 1 54 8 8 ABA HA H 4.188 0.002 1 55 8 8 ABA HB2 H 2.485 0.007 1 56 8 8 ABA HB3 H 2.539 0.002 2 57 8 8 ABA HG H 5.496 0.001 2 58 8 8 ABA CA C 55.339 0.020 1 59 8 8 ABA CB C 32.687 0.020 1 60 8 8 ABA CG C 73.710 0.020 1 61 8 8 ABA N N 119.720 0.010 1 62 9 9 ALA H H 8.050 0.001 1 63 9 9 ALA HA H 4.083 0.002 1 64 9 9 ALA HB H 1.324 0.002 1 65 9 9 ALA CA C 54.519 0.020 1 66 9 9 ALA CB C 18.622 0.020 1 67 10 10 TRP H H 7.715 0.001 1 68 10 10 TRP HA H 4.579 0.002 1 69 10 10 TRP HB2 H 3.283 0.002 2 70 10 10 TRP HB3 H 3.331 0.002 2 71 10 10 TRP HD1 H 7.317 0.002 1 72 10 10 TRP HE1 H 10.121 0.002 1 73 10 10 TRP HE3 H 7.561 0.001 1 74 10 10 TRP HH2 H 7.240 0.002 1 75 10 10 TRP HZ2 H 7.493 0.002 1 76 10 10 TRP HZ3 H 7.168 0.002 1 77 10 10 TRP CA C 56.465 0.020 1 78 10 10 TRP CB C 28.690 0.020 1 79 10 10 TRP CD1 C 127.309 0.020 1 80 10 10 TRP CE3 C 120.978 0.020 1 81 10 10 TRP CH2 C 124.847 0.020 1 82 10 10 TRP CZ2 C 114.715 0.020 1 83 10 10 TRP CZ3 C 122.333 0.020 1 84 10 10 TRP N N 118.181 0.010 1 85 10 10 TRP NE1 N 129.568 0.010 1 86 11 11 ARG H H 7.985 0.002 1 87 11 11 ARG HA H 4.030 0.002 1 88 11 11 ARG HB2 H 1.512 0.003 2 89 11 11 ARG HB3 H 1.512 0.003 2 90 11 11 ARG HD2 H 2.924 0.002 2 91 11 11 ARG HD3 H 2.924 0.002 2 92 11 11 ARG HE H 7.084 0.002 1 93 11 11 ARG HG2 H 0.965 0.002 2 94 11 11 ARG HG3 H 0.965 0.002 2 95 11 11 ARG CA C 56.414 0.020 1 96 11 11 ARG CB C 29.793 0.020 1 97 11 11 ARG CD C 43.291 0.020 1 98 11 11 ARG CG C 26.743 0.020 1 99 11 11 ARG N N 120.028 0.010 1 100 11 11 ARG NE N 84.710 0.010 1 101 12 12 CYS H H 8.193 0.002 1 102 12 12 CYS HA H 4.576 0.002 1 103 12 12 CYS HB2 H 3.180 0.002 2 104 12 12 CYS HB3 H 3.210 0.002 2 105 12 12 CYS CA C 57.182 0.020 1 106 12 12 CYS CB C 43.600 0.020 1 107 12 12 CYS N N 119.739 0.010 1 108 12 13 NH2 HN1 H 6.610 0.020 2 109 12 13 NH2 HN2 H 6.870 0.020 2 stop_ save_ ######################## # Coupling constants # ######################## save_coupling_constant_list_1 _Saveframe_category coupling_constants _Details . loop_ _Software_label $XEASY stop_ loop_ _Experiment_label '2D DQF-COSY' '2D 1H-1H eCOSY' stop_ _Sample_conditions_label $sample_conditions_1 _Spectrometer_frequency_1H 600 _Mol_system_component_name dicarba-ImI _Text_data_format . _Text_data . loop_ _Coupling_constant_ID _Coupling_constant_code _Atom_one_residue_seq_code _Atom_one_residue_label _Atom_one_name _Atom_two_residue_seq_code _Atom_two_residue_label _Atom_two_name _Coupling_constant_value _Coupling_constant_min_value _Coupling_constant_max_value _Coupling_constant_value_error 1 3JHNHA 3 CYS HN 3 CYS HA 3.5 . . 1.0 2 3JHNHA 4 SER HN 4 SER HA 10.9 . . 1.0 3 3JHNHA 7 ARG HN 7 ARG HA 7.7 . . 1.0 4 3JHNHA 8 ABA HN 8 ABA HA 8.2 . . 1.0 5 3JHNHA 10 TRP HN 10 TRP HA 4.2 . . 1.0 6 3JHNHA 11 ARG HN 11 ARG HA 4.6 . . 1.0 7 3JHNHA 12 CYS HN 12 CYS HA 6.9 . . 1.0 8 3JHAHB 3 CYS HA 3 CYS HB2 10.5 . . 2.0 9 3JHAHB 3 CYS HA 3 CYS HB3 4.1 . . 2.0 10 3JHAHB 5 ASP HA 5 ASP HB2 4.5 . . 2.0 11 3JHAHB 5 ASP HA 5 ASP HB3 10.4 . . 2.0 12 3JHAHB 10 TRP HA 10 TRP HB2 6.2 . . 2.0 13 3JHAHB 10 TRP HA 10 TRP HB3 5.5 . . 2.0 stop_ save_