data_27679 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; 1H/13C/15N Assignments for the TM domains of the KcsA potassium channel ; _BMRB_accession_number 27679 _BMRB_flat_file_name bmr27679.str _Entry_type original _Submission_date 2018-11-05 _Accession_date 2018-11-05 _Entry_origination author _NMR_STAR_version 2.1.1 _Experimental_method NMR _Details 'Assignments for the KcsA E71I mutant' loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Medeiros-Silva Joao . . 2 Jekhmane Shehrazade . . 3 Li Jing . . 4 Kummerer Felix . . 5 Muller-Hermes Christoph . . 6 Baldus Marc . . 7 Roux Benoit . . 8 Weingarth Markus . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 34 "13C chemical shifts" 79 "15N chemical shifts" 34 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2019-11-05 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 27676 'Wild Type' 27678 'E71A mutant' 27680 'E71Q mutant' stop_ _Original_release_date 2018-11-05 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Shifts in the selectivity filter dynamics cause modal gating in K+ channels ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 30631074 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Jekhmane Shehrazade . . 2 Medeiros-Silva Joao . . 3 Li Jing . . 4 Kummerer Felix . . 5 Muller-Hermes Christoph . . 6 Baldus Marc . . 7 Roux Benoit . . 8 Weingarth Markus . . stop_ _Journal_abbreviation 'Nat. Commun.' _Journal_volume 10 _Journal_issue 1 _Journal_ISSN 2041-1723 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 123 _Page_last 123 _Year 2019 _Details . loop_ _Keyword KcsA 'KcsA Mutants' 'Potassium Channel' 'Solid-State NMR' stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'KcsA Tetramer, E71I mutant' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'KcsA Monomer E71I, 1' $KcsA_E71I 'KcsA Monomer E71I, 2' $KcsA_E71I 'KcsA Monomer E71I, 3' $KcsA_E71I 'KcsA Monomer E71I, 4' $KcsA_E71I stop_ _System_molecular_weight 70776 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . loop_ _Biological_function 'Potassium Channel' stop_ _Database_query_date . _Details 'Homo-tetramer; Membrane Protein; Potassium channel' save_ ######################## # Monomeric polymers # ######################## save_KcsA_E71I _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common KcsA_E71I _Molecular_mass 17694 _Mol_thiol_state 'not present' loop_ _Biological_function 'Potassium Channel; Low-P0 mode probability' stop_ _Details 'E71I Mutant, Homo-tetramer' ############################## # Polymer residue sequence # ############################## _Residue_count 166 _Mol_residue_sequence ; MPPMLSGLLARLVKLLLGRH GSALHWRAAGAATVLLVIVL LAGSYLAVLAERGAPGAQLI TYPRALWWSVITATTVGYGD LYPVTLWGRLVAVVVMVAGI TSFGLVTAALATWFVGREQE RRGHFVRHSEKAAEEAYTRT TRALHERFDRLERMLDDNRR HHHHHH ; loop_ _Residue_seq_code _Residue_label 1 MET 2 PRO 3 PRO 4 MET 5 LEU 6 SER 7 GLY 8 LEU 9 LEU 10 ALA 11 ARG 12 LEU 13 VAL 14 LYS 15 LEU 16 LEU 17 LEU 18 GLY 19 ARG 20 HIS 21 GLY 22 SER 23 ALA 24 LEU 25 HIS 26 TRP 27 ARG 28 ALA 29 ALA 30 GLY 31 ALA 32 ALA 33 THR 34 VAL 35 LEU 36 LEU 37 VAL 38 ILE 39 VAL 40 LEU 41 LEU 42 ALA 43 GLY 44 SER 45 TYR 46 LEU 47 ALA 48 VAL 49 LEU 50 ALA 51 GLU 52 ARG 53 GLY 54 ALA 55 PRO 56 GLY 57 ALA 58 GLN 59 LEU 60 ILE 61 THR 62 TYR 63 PRO 64 ARG 65 ALA 66 LEU 67 TRP 68 TRP 69 SER 70 VAL 71 ILE 72 THR 73 ALA 74 THR 75 THR 76 VAL 77 GLY 78 TYR 79 GLY 80 ASP 81 LEU 82 TYR 83 PRO 84 VAL 85 THR 86 LEU 87 TRP 88 GLY 89 ARG 90 LEU 91 VAL 92 ALA 93 VAL 94 VAL 95 VAL 96 MET 97 VAL 98 ALA 99 GLY 100 ILE 101 THR 102 SER 103 PHE 104 GLY 105 LEU 106 VAL 107 THR 108 ALA 109 ALA 110 LEU 111 ALA 112 THR 113 TRP 114 PHE 115 VAL 116 GLY 117 ARG 118 GLU 119 GLN 120 GLU 121 ARG 122 ARG 123 GLY 124 HIS 125 PHE 126 VAL 127 ARG 128 HIS 129 SER 130 GLU 131 LYS 132 ALA 133 ALA 134 GLU 135 GLU 136 ALA 137 TYR 138 THR 139 ARG 140 THR 141 THR 142 ARG 143 ALA 144 LEU 145 HIS 146 GLU 147 ARG 148 PHE 149 ASP 150 ARG 151 LEU 152 GLU 153 ARG 154 MET 155 LEU 156 ASP 157 ASP 158 ASN 159 ARG 160 ARG 161 HIS 162 HIS 163 HIS 164 HIS 165 HIS 166 HIS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . 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 UNP P0A334 . . . . . . EMBL 1f6g 'POTASSIUM CHANNEL (KCSA)' . . . . . NCBI 1916 . . . . . . PIR S60172 'potassium channel protein' . . . . . PDB 1BL8 'POTASSIUM CHANNEL (KCSA)' . . . . . stop_ save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $KcsA_E71I 'Streptomyces lividans' 1916 Bacteria . Streptomyces lividans 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 $KcsA_E71I 'recombinant technology' . Escherichia coli M15 PQE60 stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type liposome _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $KcsA_E71I 1.5 mg '[U-100% 13C; U-100% 15N]' 'Potassium phosphate' 15 mM 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version 4.0.2 loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task collection 'data analysis' processing stop_ _Details . save_ save_SPARKY _Saveframe_category software _Name SPARKY _Version . loop_ _Vendor _Address _Electronic_address Goddard . . stop_ loop_ _Task 'chemical shift assignment' 'data analysis' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_700 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 700 _Details . save_ save_800 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 800 _Details . save_ save_950 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 950 _Details . save_ ############################# # NMR applied experiments # ############################# save_3D_HNCACB_1 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCACB' _Sample_label $sample_1 save_ save_3D_C(CO)NH_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D C(CO)NH' _Sample_label $sample_1 save_ save_3D_HNCA_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCA' _Sample_label $sample_1 save_ save_3D_HNCO_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCO' _Sample_label $sample_1 save_ save_NH_5 _Saveframe_category NMR_applied_experiment _Experiment_name NH _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 'ionic strength' 150 . mM pH 7.0 . pH temperature 273 . 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 adamantane C 13 'methyl carbon' ppm 31.48 external indirect . . . 1.0 water H 1 protons ppm 4.8 internal indirect . . . 1.0 Histidine N 15 nitrogen ppm 175.17 external indirect . . . 1.0 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_chemical_shifts_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '3D HNCACB' '3D HNCA' '3D HNCO' NH 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 'KcsA Monomer E71I, 1' _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 42 42 ALA H H 9.41 0.03 1 2 42 42 ALA C C 179.82 0.30 1 3 42 42 ALA CA C 54.87 0.30 1 4 42 42 ALA N N 121.01 0.50 1 5 43 43 GLY H H 9.19 0.03 1 6 43 43 GLY C C 175.95 0.30 1 7 43 43 GLY CA C 47.64 0.30 1 8 43 43 GLY N N 109.89 0.50 1 9 44 44 SER H H 7.61 0.03 1 10 44 44 SER C C 174.46 0.30 1 11 44 44 SER CA C 64.65 0.30 1 12 44 44 SER CB C 63.10 0.30 1 13 44 44 SER N N 115.52 0.50 1 14 45 45 TYR H H 7.07 0.03 1 15 45 45 TYR CA C 61.56 0.30 1 16 45 45 TYR N N 120.38 0.50 1 17 47 47 ALA C C 178.24 0.30 1 18 48 48 VAL H H 7.04 0.03 1 19 48 48 VAL C C 178.30 0.30 1 20 48 48 VAL CA C 65.76 0.30 1 21 48 48 VAL CB C 31.55 0.30 1 22 48 48 VAL N N 116.82 0.50 1 23 49 49 LEU H H 7.43 0.03 1 24 49 49 LEU CA C 58.18 0.30 1 25 49 49 LEU N N 117.29 0.50 1 26 53 53 GLY C C 174.47 0.30 1 27 54 54 ALA H H 7.91 0.03 1 28 54 54 ALA C C 176.96 0.30 1 29 54 54 ALA CA C 50.02 0.30 1 30 54 54 ALA N N 126.21 0.50 1 31 55 55 PRO C C 178.27 0.30 1 32 55 55 PRO CA C 64.09 0.30 1 33 55 55 PRO CB C 31.95 0.30 1 34 56 56 GLY H H 8.79 0.03 1 35 56 56 GLY N N 112.69 0.50 1 36 57 57 ALA H H 7.54 0.03 1 37 57 57 ALA CA C 54.10 0.30 1 38 57 57 ALA N N 122.33 0.50 1 39 59 59 LEU CA C 53.65 0.30 1 40 59 59 LEU CB C 39.78 0.30 1 41 60 60 ILE H H 7.04 0.03 1 42 60 60 ILE N N 106.90 0.50 1 43 61 61 THR H H 7.62 0.03 1 44 61 61 THR C C 173.58 0.30 1 45 61 61 THR CA C 59.25 0.30 1 46 61 61 THR CB C 72.15 0.30 1 47 61 61 THR N N 111.31 0.50 1 48 62 62 TYR H H 10.09 0.03 1 49 62 62 TYR C C 174.03 0.30 1 50 62 62 TYR CA C 64.16 0.30 1 51 62 62 TYR CB C 36.70 0.30 1 52 62 62 TYR N N 124.55 0.50 1 53 63 63 PRO C C 179.21 0.30 1 54 64 64 ARG H H 6.99 0.03 1 55 64 64 ARG CA C 60.31 0.30 1 56 64 64 ARG N N 113.41 0.50 1 57 65 65 ALA C C 178.63 0.30 1 58 66 66 LEU H H 8.91 0.03 1 59 66 66 LEU C C 180.09 0.30 1 60 66 66 LEU CA C 57.79 0.30 1 61 66 66 LEU N N 121.35 0.50 1 62 67 67 TRP H H 7.58 0.03 1 63 67 67 TRP HE1 H 11.79 0.03 1 64 67 67 TRP C C 178.10 0.30 1 65 67 67 TRP CA C 59.59 0.30 1 66 67 67 TRP N N 119.72 0.50 1 67 67 67 TRP NE1 N 133.06 0.50 1 68 68 68 TRP H H 8.56 0.03 1 69 68 68 TRP HE1 H 10.28 0.03 1 70 68 68 TRP C C 179.46 0.30 1 71 68 68 TRP CA C 60.94 0.30 1 72 68 68 TRP N N 118.83 0.50 1 73 68 68 TRP NE1 N 129.15 0.50 1 74 69 69 SER H H 9.77 0.03 1 75 69 69 SER C C 176.19 0.30 1 76 69 69 SER CA C 63.25 0.30 1 77 69 69 SER N N 122.79 0.50 1 78 70 70 VAL H H 7.48 0.03 1 79 70 70 VAL CA C 66.88 0.30 1 80 70 70 VAL N N 123.78 0.50 1 81 72 72 THR C C 174.81 0.30 1 82 72 72 THR CA C 67.45 0.30 1 83 73 73 ALA H H 8.39 0.03 1 84 73 73 ALA C C 176.22 0.30 1 85 73 73 ALA CA C 55.75 0.30 1 86 73 73 ALA N N 123.81 0.50 1 87 74 74 THR H H 6.95 0.03 1 88 74 74 THR C C 176.38 0.30 1 89 74 74 THR CA C 61.77 0.30 1 90 74 74 THR CB C 70.60 0.30 1 91 74 74 THR N N 99.26 0.50 1 92 75 75 THR H H 7.31 0.03 1 93 75 75 THR C C 172.86 0.30 1 94 75 75 THR CA C 63.80 0.30 1 95 75 75 THR CB C 69.23 0.30 1 96 75 75 THR N N 110.27 0.50 1 97 76 76 VAL H H 7.19 0.03 1 98 76 76 VAL C C 179.86 0.30 1 99 76 76 VAL CA C 66.31 0.30 1 100 76 76 VAL N N 122.66 0.50 1 101 77 77 GLY H H 7.78 0.03 1 102 77 77 GLY C C 175.06 0.30 1 103 77 77 GLY CA C 48.55 0.30 1 104 77 77 GLY N N 101.99 0.50 1 105 78 78 TYR H H 5.85 0.03 1 106 78 78 TYR C C 178.13 0.30 1 107 78 78 TYR CA C 60.23 0.30 1 108 78 78 TYR CB C 39.08 0.30 1 109 78 78 TYR N N 114.24 0.50 1 110 79 79 GLY H H 9.32 0.03 1 111 79 79 GLY C C 174.20 0.30 1 112 79 79 GLY CA C 45.76 0.30 1 113 79 79 GLY N N 101.29 0.50 1 114 80 80 ASP H H 9.86 0.03 1 115 80 80 ASP C C 179.40 0.30 1 116 80 80 ASP CA C 56.25 0.30 1 117 80 80 ASP CB C 36.98 0.30 1 118 80 80 ASP CG C 176.16 0.30 1 119 80 80 ASP N N 119.56 0.50 1 120 81 81 LEU H H 7.01 0.03 1 121 81 81 LEU C C 176.20 0.30 1 122 81 81 LEU CA C 53.30 0.30 1 123 81 81 LEU CB C 46.85 0.30 1 124 81 81 LEU CG C 27.45 0.30 1 125 81 81 LEU N N 117.20 0.50 1 126 82 82 TYR H H 7.74 0.03 1 127 82 82 TYR C C 172.13 0.30 1 128 82 82 TYR CA C 55.45 0.30 1 129 82 82 TYR CB C 35.34 0.30 1 130 82 82 TYR N N 115.01 0.50 1 131 83 83 PRO C C 176.81 0.30 1 132 84 84 VAL H H 11.09 0.03 1 133 84 84 VAL C C 176.46 0.30 1 134 84 84 VAL CA C 60.95 0.30 1 135 84 84 VAL CB C 32.77 0.30 1 136 84 84 VAL N N 119.18 0.50 1 137 85 85 THR H H 9.27 0.03 1 138 85 85 THR C C 175.41 0.30 1 139 85 85 THR CA C 61.13 0.30 1 140 85 85 THR CB C 72.74 0.30 1 141 85 85 THR N N 117.40 0.50 1 142 86 86 LEU H H 8.39 0.03 1 143 86 86 LEU CA C 59.05 0.30 1 144 86 86 LEU N N 122.50 0.50 1 145 87 87 TRP H H 7.47 0.03 1 146 87 87 TRP CA C 59.88 0.30 1 147 87 87 TRP N N 115.75 0.50 1 stop_ save_