data_19030 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Analysis of the structural and molecular basis of voltage-sensitive sodium channel inhibition by the spider toxin, Huwentoxin-IV ( -TRTX-Hh2a). ; _BMRB_accession_number 19030 _BMRB_flat_file_name bmr19030.str _Entry_type original _Submission_date 2013-02-12 _Accession_date 2013-02-12 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details HWTX-IV(W30A) loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Gibbs Al . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 167 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2013-08-07 update BMRB 'update entry citation' 2013-06-17 original author 'original release' stop_ loop_ _Related_BMRB_accession_number _Relationship 19026 HWTX-IV 19032 Huwentoxin-IV stop_ save_ ############################# # Citation for this entry # ############################# save_citations _Saveframe_category entry_citation _Citation_full . _Citation_title 'Analysis of the Structural and Molecular Basis of Voltage-sensitive Sodium Channel Inhibition by the Spider Toxin Huwentoxin-IV (-TRTX-Hh2a).' _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 23760503 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Minassian Natali A. . 2 Gibbs Alan . . 3 Shih Amy Y. . 4 Liu Yi . . 5 Neff Robert A. . 6 Sutton Steven W. . 7 Mirzadegan Tara . . 8 Connor Judith . . 9 Fellows Ross . . 10 Husovsky Matthew . . 11 Nelson Serena . . 12 Hunter Michael J. . 13 Flinspach Mack . . 14 Wickenden Alan D. . stop_ _Journal_abbreviation 'J. Biol. Chem.' _Journal_name_full 'The Journal of biological chemistry' _Journal_volume 288 _Journal_issue 31 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 22707 _Page_last 22720 _Year 2013 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name Huwentoxin-IV _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label Huwentoxin-IV $Huwentoxin-IV 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_Huwentoxin-IV _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Huwentoxin-IV _Molecular_mass 4005.742 _Mol_thiol_state . _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 35 _Mol_residue_sequence ; ECLEIFKACNPSNDQCCKSS KLVCSRKTRACKYQI ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 GLU 2 2 CYS 3 3 LEU 4 4 GLU 5 5 ILE 6 6 PHE 7 7 LYS 8 8 ALA 9 9 CYS 10 10 ASN 11 11 PRO 12 12 SER 13 13 ASN 14 14 ASP 15 15 GLN 16 16 CYS 17 17 CYS 18 18 LYS 19 19 SER 20 20 SER 21 21 LYS 22 22 LEU 23 23 VAL 24 24 CYS 25 25 SER 26 26 ARG 27 27 LYS 28 28 THR 29 29 ARG 30 30 ALA 31 31 CYS 32 32 LYS 33 33 TYR 34 34 GLN 35 35 ILE stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date 2015-10-14 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 19026 HWTX-IV 100.00 35 97.14 97.14 5.18e-14 PDB 1MB6 "Three Dimensional Solution Structure Of Huwentoxin-Iv By 2d 1h-Nmr" 100.00 35 97.14 97.14 5.18e-14 PDB 2M4X "Analysis Of The Structural And Molecular Basis Of Voltage-sensitive Sodium Channel Inhibition By The Spider Toxin, Huwentoxin-i" 100.00 35 97.14 97.14 5.18e-14 PDB 2M4Z "Analysis Of The Structural And Molecular Basis Of Voltage-sensitive Sodium Channel Inhibition By The Spider Toxin, Huwentoxin-i" 100.00 35 100.00 100.00 8.78e-15 GB AAP33074 "huwentoxin-IV precursor [Haplopelma schmidti]" 100.00 89 97.14 97.14 1.24e-14 GB ABY77744 "HWTX-IVa precursor [Haplopelma schmidti]" 100.00 86 97.14 97.14 1.44e-14 GB ABY77746 "HWTX-IVc precursor [Haplopelma schmidti]" 100.00 86 97.14 97.14 1.42e-14 SP P83303 "RecName: Full=Mu-theraphotoxin-Hs2a; Short=Mu-TRTX-Hs2a; AltName: Full=Huwentoxin-4; AltName: Full=Huwentoxin-IV; Short=HwTx-IV" 100.00 89 97.14 97.14 1.24e-14 stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $Huwentoxin-IV . . . . . . 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 $Huwentoxin-IV 'recombinant technology' . Homo Sapien . pCDNA 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 $Huwentoxin-IV 2 mM 'natural abundance' 'sodium phosphate' 20 mM 'natural abundance' 'sodium azide' 150 uM 'natural abundance' EDTA 100 uM [U-2H] H2O 90 % 'natural abundance' D2O 10 % '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_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 950 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_1H-15N_HSQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_1 save_ save_2D_1H-1H_TOCSY_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H TOCSY' _Sample_label $sample_1 save_ save_2D_1H-1H_NOESY_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-1H NOESY' _Sample_label $sample_1 save_ save_2D_1H-13C_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C HSQC' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details . loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units temperature 298 . K pH 6.7 . pH pressure 1 . atm 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 4.7 internal direct . . . 1 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-1H TOCSY' '2D 1H-1H NOESY' '2D 1H-13C HSQC' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name Huwentoxin-IV _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 CYS H H 8.262 0.000 1 2 2 2 CYS HA H 4.982 0.000 1 3 2 2 CYS HB2 H 3.217 0.000 2 4 2 2 CYS HB3 H 3.363 0.000 2 5 3 3 LEU H H 8.712 0.000 1 6 3 3 LEU HA H 4.426 0.000 1 7 3 3 LEU HB2 H 1.588 0.000 2 8 3 3 LEU HB3 H 1.716 0.000 2 9 3 3 LEU HG H 1.377 0.000 1 10 3 3 LEU HD1 H 0.998 0.000 1 11 3 3 LEU HD2 H 0.881 0.000 1 12 4 4 GLU H H 7.833 0.000 1 13 4 4 GLU HA H 4.145 0.000 1 14 4 4 GLU HB2 H 2.265 0.000 2 15 4 4 GLU HG2 H 2.397 0.000 2 16 4 4 GLU HG3 H 2.484 0.000 2 17 5 5 ILE H H 7.910 0.000 1 18 5 5 ILE HA H 3.160 0.000 1 19 5 5 ILE HB H 1.362 0.000 1 20 5 5 ILE HG12 H -0.062 0.000 2 21 5 5 ILE HG13 H 0.663 0.000 2 22 5 5 ILE HG2 H 0.294 0.000 1 23 5 5 ILE HD1 H 0.245 0.000 1 24 6 6 PHE H H 8.035 0.000 1 25 6 6 PHE HA H 4.208 0.000 1 26 6 6 PHE HB2 H 3.473 0.000 2 27 6 6 PHE HB3 H 3.554 0.000 2 28 6 6 PHE HE1 H 7.306 0.000 1 29 6 6 PHE HE2 H 7.306 0.000 1 30 7 7 LYS H H 7.824 0.000 1 31 7 7 LYS HA H 4.346 0.000 1 32 7 7 LYS HD2 H 1.837 0.000 2 33 7 7 LYS HD3 H 2.143 0.000 2 34 8 8 ALA H H 8.296 0.000 1 35 8 8 ALA HA H 4.753 0.000 1 36 8 8 ALA HB H 1.497 0.000 1 37 9 9 CYS H H 8.497 0.000 1 38 9 9 CYS HA H 4.827 0.000 1 39 9 9 CYS HB2 H 2.948 0.000 2 40 9 9 CYS HB3 H 3.096 0.000 2 41 10 10 ASN H H 9.168 0.000 1 42 10 10 ASN HA H 5.178 0.000 1 43 10 10 ASN HB2 H 2.798 0.000 2 44 10 10 ASN HB3 H 3.048 0.000 2 45 10 10 ASN HD21 H 7.840 0.000 2 46 10 10 ASN HD22 H 7.163 0.000 2 47 11 11 PRO HA H 3.940 0.000 1 48 11 11 PRO HB2 H 1.945 0.000 2 49 11 11 PRO HB3 H 1.945 0.000 2 50 11 11 PRO HG2 H 1.819 0.000 2 51 11 11 PRO HG3 H 1.854 0.000 2 52 11 11 PRO HD2 H 3.275 0.000 2 53 11 11 PRO HD3 H 3.275 0.000 2 54 12 12 SER H H 7.850 0.000 1 55 12 12 SER HA H 4.468 0.000 1 56 12 12 SER HB2 H 3.848 0.000 2 57 12 12 SER HB3 H 3.970 0.000 2 58 13 13 ASN H H 7.671 0.000 1 59 13 13 ASN HA H 4.678 0.000 1 60 13 13 ASN HB2 H 2.643 0.000 2 61 13 13 ASN HB3 H 2.677 0.000 2 62 14 14 ASP H H 8.755 0.000 1 63 14 14 ASP HA H 4.122 0.000 1 64 14 14 ASP HB2 H 2.526 0.000 2 65 14 14 ASP HB3 H 3.044 0.000 2 66 15 15 GLN H H 8.113 0.000 1 67 15 15 GLN HA H 4.636 0.000 1 68 15 15 GLN HB2 H 2.359 0.000 2 69 15 15 GLN HB3 H 2.359 0.000 2 70 15 15 GLN HG2 H 2.764 0.000 2 71 15 15 GLN HG3 H 2.438 0.000 2 72 16 16 CYS H H 9.300 0.000 1 73 16 16 CYS HA H 4.900 0.000 1 74 16 16 CYS HB2 H 2.639 0.000 2 75 16 16 CYS HB3 H 2.719 0.000 2 76 17 17 CYS H H 9.384 0.000 1 77 17 17 CYS HA H 4.549 0.000 1 78 17 17 CYS HB2 H 3.194 0.000 2 79 17 17 CYS HB3 H 2.783 0.000 2 80 18 18 LYS H H 9.050 0.000 1 81 18 18 LYS HA H 4.251 0.000 1 82 18 18 LYS HB2 H 1.919 0.000 2 83 18 18 LYS HB3 H 1.959 0.000 2 84 18 18 LYS HD2 H 1.633 0.000 2 85 18 18 LYS HD3 H 1.705 0.000 2 86 19 19 SER H H 9.073 0.000 1 87 19 19 SER HA H 4.289 0.000 1 88 19 19 SER HB2 H 3.964 0.000 2 89 19 19 SER HB3 H 4.047 0.000 2 90 20 20 SER H H 6.796 0.000 1 91 20 20 SER HA H 4.719 0.000 1 92 20 20 SER HB2 H 3.674 0.000 2 93 20 20 SER HB3 H 4.174 0.000 2 94 21 21 LYS H H 8.063 0.000 1 95 21 21 LYS HA H 3.880 0.000 1 96 21 21 LYS HB2 H 2.214 0.000 2 97 21 21 LYS HB3 H 1.991 0.000 2 98 22 22 LEU H H 6.978 0.000 1 99 22 22 LEU HA H 5.148 0.000 1 100 22 22 LEU HB2 H 1.756 0.000 2 101 22 22 LEU HB3 H 1.756 0.000 2 102 22 22 LEU HG H 1.434 0.000 1 103 22 22 LEU HD1 H 0.813 0.000 1 104 22 22 LEU HD2 H 0.813 0.000 1 105 23 23 VAL H H 9.164 0.000 1 106 23 23 VAL HA H 4.352 0.000 1 107 23 23 VAL HB H 1.893 0.000 1 108 23 23 VAL HG1 H 0.866 0.000 1 109 23 23 VAL HG2 H 0.798 0.000 1 110 24 24 CYS H H 9.637 0.000 1 111 24 24 CYS HA H 4.541 0.000 1 112 24 24 CYS HB2 H 3.128 0.000 2 113 24 24 CYS HB3 H 2.552 0.000 2 114 25 25 SER H H 7.843 0.000 1 115 25 25 SER HA H 4.466 0.000 1 116 25 25 SER HB2 H 3.857 0.000 2 117 25 25 SER HB3 H 3.975 0.000 2 118 26 26 ARG H H 9.127 0.000 1 119 26 26 ARG HA H 3.950 0.000 1 120 26 26 ARG HB2 H 1.861 0.000 2 121 26 26 ARG HB3 H 1.944 0.000 2 122 26 26 ARG HG2 H 1.686 0.000 2 123 26 26 ARG HG3 H 1.831 0.000 2 124 27 27 LYS H H 7.898 0.000 1 125 27 27 LYS HA H 4.433 0.000 1 126 28 28 THR H H 7.706 0.000 1 127 28 28 THR HA H 4.413 0.000 1 128 28 28 THR HB H 4.378 0.000 1 129 28 28 THR HG2 H 1.197 0.000 1 130 29 29 ARG H H 7.413 0.000 1 131 29 29 ARG HA H 3.838 0.000 1 132 29 29 ARG HB2 H 2.073 0.000 2 133 29 29 ARG HB3 H 2.371 0.000 2 134 29 29 ARG HG2 H 1.350 0.000 2 135 29 29 ARG HG3 H 1.556 0.000 2 136 29 29 ARG HD2 H 3.248 0.000 2 137 29 29 ARG HD3 H 3.342 0.000 2 138 30 30 ALA H H 7.349 0.000 1 139 30 30 ALA HA H 5.248 0.000 1 140 30 30 ALA HB H 1.095 0.000 1 141 31 31 CYS H H 8.573 0.000 1 142 31 31 CYS HA H 4.900 0.000 1 143 31 31 CYS HB2 H 3.047 0.000 2 144 31 31 CYS HB3 H 2.497 0.000 2 145 32 32 LYS H H 9.543 0.000 1 146 32 32 LYS HA H 4.625 0.000 1 147 32 32 LYS HB2 H 1.771 0.000 2 148 32 32 LYS HB3 H 1.909 0.000 2 149 32 32 LYS HG2 H 1.364 0.000 2 150 32 32 LYS HG3 H 1.364 0.000 2 151 32 32 LYS HD2 H 1.531 0.000 2 152 32 32 LYS HD3 H 1.531 0.000 2 153 33 33 TYR H H 8.391 0.000 1 154 33 33 TYR HA H 4.714 0.000 1 155 33 33 TYR HB2 H 2.793 0.000 2 156 33 33 TYR HB3 H 3.077 0.000 2 157 34 34 GLN H H 8.784 0.000 1 158 34 34 GLN HA H 4.217 0.000 1 159 34 34 GLN HB2 H 1.844 0.000 2 160 34 34 GLN HB3 H 1.844 0.000 2 161 34 34 GLN HG2 H 2.255 0.000 2 162 34 34 GLN HG3 H 2.255 0.000 2 163 35 35 ILE H H 8.148 0.000 1 164 35 35 ILE HA H 4.185 0.000 1 165 35 35 ILE HB H 1.866 0.000 1 166 35 35 ILE HG12 H 1.216 0.000 2 167 35 35 ILE HD1 H 0.953 0.000 1 stop_ save_