data_30645 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Influenza B M2 Proton Channel in the Closed State - SSNMR Structure at pH 7.5 ; _BMRB_accession_number 30645 _BMRB_flat_file_name bmr30645.str _Entry_type original _Submission_date 2019-07-21 _Accession_date 2019-07-21 _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 Mandala V. S. . 2 Loftis A. R. . 3 Shcherbakov A. S. . 4 Pentelute B. L. . 5 Hong M. . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "13C chemical shifts" 171 "15N chemical shifts" 27 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2020-01-30 original BMRB . stop_ _Original_release_date 2019-07-29 save_ ############################# # Citation for this entry # ############################# save_citation_1 _Saveframe_category entry_citation _Citation_full . _Citation_title ; Atomic Structures of a Closed and Open Proton Channel in Lipid Bilayers Reveal the Asymmetric Conduction Mechanism ; _Citation_status 'in preparation' _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID ? loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Mandala V. S. . 2 Loftis A. R. . 3 Shcherbakov A. S. . 4 Pentelute B. L. . 5 Hong M. . . stop_ _Journal_abbreviation . _Journal_volume . _Journal_issue . _Journal_CSD 0353 _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year . _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'BM2 protein' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'entity_1, 1' $entity_1 'entity_1, 2' $entity_1 'entity_1, 3' $entity_1 'entity_1, 4' $entity_1 stop_ _System_molecular_weight . _System_oligomer_state ? _System_paramagnetic no _System_thiol_state 'all free' _Database_query_date . _Details . save_ ######################## # Monomeric polymers # ######################## save_entity_1 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common entity_1 _Molecular_mass 5953.146 _Mol_thiol_state 'all free' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 51 _Mol_residue_sequence ; MFEPFQILSICSFILSALHF MAWTIGHLNQIKRGVNMKIR IKGPNKETINR ; loop_ _Residue_seq_code _Residue_label 1 MET 2 PHE 3 GLU 4 PRO 5 PHE 6 GLN 7 ILE 8 LEU 9 SER 10 ILE 11 CYS 12 SER 13 PHE 14 ILE 15 LEU 16 SER 17 ALA 18 LEU 19 HIS 20 PHE 21 MET 22 ALA 23 TRP 24 THR 25 ILE 26 GLY 27 HIS 28 LEU 29 ASN 30 GLN 31 ILE 32 LYS 33 ARG 34 GLY 35 VAL 36 ASN 37 MET 38 LYS 39 ILE 40 ARG 41 ILE 42 LYS 43 GLY 44 PRO 45 ASN 46 LYS 47 GLU 48 THR 49 ILE 50 ASN 51 ARG stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species _Strain _Gene_mnemonic $entity_1 'Influenza B virus' 1601064 Viruses . Betainfluenzavirus 'Influenza B virus' B/Maryland/1/2001 BM2 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 $entity_1 'recombinant technology' . . . BL21(DE3) Plasmid pET-SUMO stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type membrane _Details '0.26 mg/uL [U-13C; U-15N] Influenza B M2 Proton Channel, 0.48 mg/uL POPE, aqueous buffer' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $entity_1 0.26 mg/uL '[U-13C; U-15N]' POPE 0.48 mg/uL 'natural abundance' stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type membrane _Details '0.13 mg/uL [U-13C; U-15N] and 4-19F-F5, 4-19F-F20 Influenza B M2 Proton Channel, 0.39 mg/uL POPE, aqueous buffer' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $entity_1 0.13 mg/uL '[U-13C; U-15N] and 4-19F-F5, 4-19F-F20' POPE 0.39 mg/uL 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_software_1 _Saveframe_category software _Name 'X-PLOR NIH' _Version 2.47 loop_ _Vendor _Address _Electronic_address 'Schwieters, Kuszewski, Tjandra and Clore' . . stop_ loop_ _Task refinement 'structure calculation' stop_ _Details . save_ save_software_2 _Saveframe_category software _Name Sparky _Version 3.1 loop_ _Vendor _Address _Electronic_address Goddard . . stop_ loop_ _Task 'chemical shift assignment' 'peak picking' stop_ _Details . save_ save_software_3 _Saveframe_category software _Name TopSpin _Version 2.1-3.6 loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task processing stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model 'AVANCE II' _Field_strength 800 _Details . save_ save_NMR_spectrometer_2 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model 'AVANCE III' _Field_strength 600 _Details . save_ ############################# # NMR applied experiments # ############################# save_2D_CORD_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D CORD' _Sample_label $sample_1 save_ save_2D_NC_TEDOR_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D NC TEDOR' _Sample_label $sample_1 save_ save_3D_NCCX_TEDOR-CORD_3 _Saveframe_category NMR_applied_experiment _Experiment_name '3D NCCX TEDOR-CORD' _Sample_label $sample_1 save_ save_3D_CCC_CORD-CORD_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D CCC CORD-CORD' _Sample_label $sample_1 save_ save_2D_NC_NH-DIPSHIFT_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D NC NH-DIPSHIFT' _Sample_label $sample_1 save_ save_2D_CC_Water-edited_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D CC Water-edited' _Sample_label $sample_1 save_ save_1D_13C-19F_REDOR_7 _Saveframe_category NMR_applied_experiment _Experiment_name '1D 13C-19F REDOR' _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 'ionic strength' 20 0.02 mM pH 7.5 0.2 pH pressure 1 0.05 atm temperature 285 5 K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chem_shift_reference_1 _Saveframe_category chemical_shift_reference _Details ; 13C chemical shifts were referenced to the tetramethylsilane (TMS) scale and 15N chemical shifts were referenced to the liquid ammonia scale. ; 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 carbons' ppm 38.48 external direct . . . 0.25 NAV N 15 nitrogen ppm 122.00 external direct . . . 0.1 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_chemical_shifts_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D CORD' '2D NC TEDOR' '3D NCCX TEDOR-CORD' '3D CCC CORD-CORD' '2D NC NH-DIPSHIFT' '2D CC Water-edited' '1D 13C-19F REDOR' stop_ loop_ _Sample_label $sample_1 $sample_2 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chem_shift_reference_1 _Mol_system_component_name 'entity_1, 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 2 2 PHE CA C 56.5 0.5 1 2 2 2 PHE CB C 37.9 0.5 1 3 3 3 GLU C C 174.6 0.5 1 4 3 3 GLU CA C 52.5 0.5 1 5 3 3 GLU CB C 27.3 0.5 1 6 3 3 GLU CG C 34.1 0.5 1 7 3 3 GLU CD C 181.6 0.5 1 8 3 3 GLU N N 121 1.0 1 9 4 4 PRO C C 176.1 0.5 1 10 4 4 PRO CA C 65.2 0.5 1 11 4 4 PRO CB C 30.6 0.5 1 12 4 4 PRO CG C 26.3 0.5 1 13 4 4 PRO CD C 48.5 0.5 1 14 4 4 PRO N N 137.1 1.0 1 15 5 5 PHE CA C 57 0.5 1 16 5 5 PHE CB C 35.3 0.5 1 17 5 5 PHE N N 121.4 1.0 1 18 6 6 GLN CA C 53.6 0.5 1 19 6 6 GLN CB C 27.3 0.5 1 20 6 6 GLN CG C 32.2 0.5 1 21 6 6 GLN CD C 178.5 0.5 1 22 7 7 ILE C C 176 0.5 1 23 7 7 ILE CA C 63.9 0.5 1 24 7 7 ILE CB C 36 0.5 1 25 7 7 ILE CG1 C 27.6 0.5 1 26 7 7 ILE CG2 C 15.8 0.5 1 27 7 7 ILE CD1 C 12.1 0.5 1 28 7 7 ILE N N 122 1.0 1 29 8 8 LEU C C 176.4 0.5 1 30 8 8 LEU CA C 56.6 0.5 1 31 8 8 LEU CB C 41.2 0.5 1 32 8 8 LEU CG C 25.9 0.5 1 33 8 8 LEU CD1 C 24.3 0.5 1 34 8 8 LEU CD2 C 22.3 0.5 1 35 8 8 LEU N N 120.8 1.0 1 36 9 9 SER C C 177 0.5 1 37 9 9 SER CA C 60.7 0.5 1 38 9 9 SER CB C 60.5 0.5 1 39 9 9 SER N N 115.9 1.0 1 40 10 10 ILE C C 175.7 0.5 1 41 10 10 ILE CA C 63.7 0.5 1 42 10 10 ILE CB C 36.3 0.5 1 43 10 10 ILE CG1 C 27.6 0.5 1 44 10 10 ILE CG2 C 15.6 0.5 1 45 10 10 ILE CD1 C 12.2 0.5 1 46 10 10 ILE N N 118.7 1.0 1 47 11 11 CYS C C 175.1 0.5 1 48 11 11 CYS CA C 63.1 0.5 1 49 11 11 CYS CB C 25 0.5 1 50 11 11 CYS N N 117.7 1.0 1 51 12 12 SER C C 175.6 0.5 1 52 12 12 SER CA C 60.3 0.5 1 53 12 12 SER CB C 61.4 0.5 1 54 12 12 SER N N 116.2 1.0 1 55 13 13 PHE C C 176.1 0.5 1 56 13 13 PHE CA C 60 0.5 1 57 13 13 PHE CB C 36.6 0.5 1 58 13 13 PHE CD1 C 129.1 0.5 3 59 13 13 PHE CD2 C 129.1 0.5 3 60 13 13 PHE CE1 C 128.9 0.5 3 61 13 13 PHE CE2 C 128.9 0.5 3 62 13 13 PHE N N 119.7 1.0 1 63 14 14 ILE C C 176.3 0.5 1 64 14 14 ILE CA C 63.4 0.5 1 65 14 14 ILE CB C 36.1 0.5 1 66 14 14 ILE CG1 C 27 0.5 1 67 14 14 ILE CG2 C 15.8 0.5 1 68 14 14 ILE CD1 C 11.7 0.5 1 69 14 14 ILE N N 118.9 1.0 1 70 15 15 LEU C C 177 0.5 1 71 15 15 LEU CA C 56.6 0.5 1 72 15 15 LEU CB C 40.2 0.5 1 73 15 15 LEU CG C 25.1 0.5 1 74 15 15 LEU CD1 C 24.8 0.5 1 75 15 15 LEU CD2 C 23.1 0.5 1 76 15 15 LEU N N 120 1.0 1 77 16 16 SER C C 175.3 0.5 1 78 16 16 SER CA C 59.9 0.5 1 79 16 16 SER CB C 60.7 0.5 1 80 16 16 SER N N 116 1.0 1 81 17 17 ALA C C 176.7 0.5 1 82 17 17 ALA CA C 53.6 0.5 1 83 17 17 ALA CB C 15.8 0.5 1 84 17 17 ALA N N 127.3 1.0 1 85 18 18 LEU C C 176.4 0.5 1 86 18 18 LEU CA C 56.3 0.5 1 87 18 18 LEU CB C 39.9 0.5 1 88 18 18 LEU CG C 24.7 0.5 1 89 18 18 LEU CD1 C 24.7 0.5 1 90 18 18 LEU CD2 C 21.5 0.5 1 91 18 18 LEU N N 118.1 1.0 1 92 19 19 HIS C C 175.6 0.5 1 93 19 19 HIS CA C 55.6 0.5 1 94 19 19 HIS CB C 30 0.5 1 95 19 19 HIS CG C 135.3 0.5 1 96 19 19 HIS CD2 C 114.4 0.5 1 97 19 19 HIS CE1 C 136 0.5 1 98 19 19 HIS N N 119.2 1.0 1 99 20 20 PHE C C 174.4 0.5 1 100 20 20 PHE CA C 59.7 0.5 1 101 20 20 PHE CB C 36.5 0.5 1 102 20 20 PHE CD1 C 129 0.5 3 103 20 20 PHE CD2 C 129 0.5 3 104 20 20 PHE CE1 C 129 0.5 3 105 20 20 PHE CE2 C 129 0.5 3 106 20 20 PHE N N 120.9 1.0 1 107 21 21 MET C C 175.8 0.5 1 108 21 21 MET CA C 58.1 0.5 1 109 21 21 MET CB C 30.8 0.5 1 110 21 21 MET CG C 32.4 0.5 1 111 21 21 MET N N 121.3 1.0 1 112 22 22 ALA C C 176.7 0.5 1 113 22 22 ALA CA C 53.7 0.5 1 114 22 22 ALA CB C 16.4 0.5 1 115 22 22 ALA N N 123.2 1.0 1 116 23 23 TRP C C 175.4 0.5 1 117 23 23 TRP CA C 60 0.5 1 118 23 23 TRP CB C 26.3 0.5 1 119 23 23 TRP CG C 111.8 0.5 1 120 23 23 TRP CD1 C 122.2 0.5 1 121 23 23 TRP CD2 C 128.3 0.5 1 122 23 23 TRP CE2 C 137 0.5 1 123 23 23 TRP N N 119.6 1.0 1 124 23 23 TRP NE1 N 130.8 1.0 1 125 24 24 THR C C 174.4 0.5 1 126 24 24 THR CA C 66 0.5 1 127 24 24 THR CB C 65.9 0.5 1 128 24 24 THR CG2 C 19.7 0.5 1 129 24 24 THR N N 117.8 1.0 1 130 25 25 ILE C C 176.5 0.5 1 131 25 25 ILE CA C 63.4 0.5 1 132 25 25 ILE CB C 36 0.5 1 133 25 25 ILE CG1 C 26.9 0.5 1 134 25 25 ILE CG2 C 15.8 0.5 1 135 25 25 ILE CD1 C 12.3 0.5 1 136 25 25 ILE N N 121.8 1.0 1 137 26 26 GLY C C 174.2 0.5 1 138 26 26 GLY CA C 45.5 0.5 1 139 26 26 GLY N N 107.5 1.0 1 140 27 27 HIS C C 175.9 0.5 1 141 27 27 HIS CA C 58 0.5 1 142 27 27 HIS CB C 28.3 0.5 1 143 27 27 HIS CG C 135.9 0.5 1 144 27 27 HIS CD2 C 114.5 0.5 1 145 27 27 HIS CE1 C 135.8 0.5 1 146 27 27 HIS N N 121.6 1.0 1 147 28 28 LEU CA C 55.8 0.5 1 148 28 28 LEU CB C 39.7 0.5 1 149 29 29 ASN C C 174 0.5 1 150 29 29 ASN CA C 51.5 0.5 1 151 29 29 ASN CB C 37.1 0.5 1 152 29 29 ASN N N 123.6 1.0 1 153 32 32 LYS CB C 30.9 0.5 1 154 32 32 LYS CD C 27.4 0.5 1 155 32 32 LYS CE C 40.3 0.5 1 156 32 32 LYS NZ N 33.7 1.0 1 157 33 33 ARG CA C 52.4 0.5 1 158 33 33 ARG CB C 29 0.5 1 159 33 33 ARG CG C 25.8 0.5 1 160 33 33 ARG CD C 41.6 0.5 1 161 33 33 ARG CZ C 157.5 0.5 1 162 34 34 GLY C C 172.9 0.5 1 163 34 34 GLY CA C 44.4 0.5 1 164 35 35 VAL CA C 59.7 0.5 1 165 35 35 VAL CB C 30.7 0.5 1 166 35 35 VAL CG1 C 19.6 0.5 2 167 35 35 VAL CG2 C 19.6 0.5 2 168 43 43 GLY C C 169.8 0.5 1 169 43 43 GLY CA C 42.7 0.5 1 170 44 44 PRO C C 174.4 0.5 1 171 44 44 PRO CA C 61 0.5 1 172 44 44 PRO CB C 30.5 0.5 1 173 44 44 PRO CG C 25.3 0.5 1 174 44 44 PRO CD C 48.3 0.5 1 175 45 45 ASN CA C 51.5 0.5 1 176 45 45 ASN CB C 37.1 0.5 1 177 46 46 LYS CA C 54.6 0.5 1 178 46 46 LYS CB C 31.1 0.5 1 179 46 46 LYS CG C 22.9 0.5 1 180 46 46 LYS CD C 27.3 0.5 1 181 46 46 LYS CE C 40.2 0.5 1 182 47 47 GLU CA C 54.8 0.5 1 183 47 47 GLU CB C 28.5 0.5 1 184 47 47 GLU CG C 34.5 0.5 1 185 48 48 THR CA C 60.2 0.5 1 186 48 48 THR CB C 67.9 0.5 1 187 48 48 THR CG2 C 19.9 0.5 1 188 49 49 ILE CA C 59.2 0.5 1 189 49 49 ILE CB C 37 0.5 1 190 49 49 ILE CG1 C 25.4 0.5 1 191 49 49 ILE CG2 C 15.7 0.5 1 192 49 49 ILE CD1 C 11.2 0.5 1 193 50 50 ASN CA C 51.4 0.5 1 194 50 50 ASN CB C 37.1 0.5 1 195 51 51 ARG CA C 55.6 0.5 1 196 51 51 ARG CB C 29.8 0.5 1 197 51 51 ARG CG C 25.4 0.5 1 198 51 51 ARG CD C 41.7 0.5 1 stop_ save_