data_27316 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; NMR assignment of the transmembrane helix of BclxL in phospholipid nanodiscs ; _BMRB_accession_number 27316 _BMRB_flat_file_name bmr27316.str _Entry_type original _Submission_date 2017-11-24 _Accession_date 2017-11-24 _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 Raltchev Kolio . . 2 Pipercevic Joka . . 3 Hagn Franz . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 26 "13C chemical shifts" 62 "15N chemical shifts" 26 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2018-07-12 original BMRB . stop_ _Original_release_date 2017-11-27 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Structural analysis of natively-folded membrane-anchored proteins obtained by SortaseA-mediated ligation ; _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 Raltchev Kolio . . 2 Pipercevic Joka . . 3 Hagn Franz . . stop_ _Journal_abbreviation 'Not known' _Journal_volume . _Journal_issue . _Journal_CSD . _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 BclxL _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label BclxL $BclxL 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_BclxL _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common BclxL _Molecular_mass . _Mol_thiol_state 'not present' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 35 _Mol_residue_sequence ; GSGESRKGQERFNRWFLTGM TVAGVVLLGSLFSRK ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 209 GLY 2 210 SER 3 211 GLY 4 212 GLU 5 213 SER 6 214 ARG 7 215 LYS 8 216 GLY 9 217 GLN 10 218 GLU 11 219 ARG 12 220 PHE 13 221 ASN 14 222 ARG 15 223 TRP 16 224 PHE 17 225 LEU 18 226 THR 19 227 GLY 20 228 MET 21 229 THR 22 230 VAL 23 231 ALA 24 232 GLY 25 233 VAL 26 234 VAL 27 235 LEU 28 236 LEU 29 237 GLY 30 238 SER 31 239 LEU 32 240 PHE 33 241 SER 34 242 ARG 35 243 LYS 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 $BclxL Human 9606 Eukaryota Metazoa Homo sapiens stop_ save_ ######################### # Experimental source # ######################### save_experimental_source _Saveframe_category experimental_source loop_ _Mol_label _Production_method _Host_organism_name_common _Genus _Species _Strain _Variant _Vector_name _Details $BclxL 'recombinant technology' . Escherichia coli K12 BL(DE3) pET21 'N-terminal GB1 expression tag and Thrombin cleavage site' stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type solution _Details 'BclxL TM helix in phospholipid nanodsics' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling $BclxL 400 mM '[U-13C; U-15N; U-2H]' DMPC 15 mM [U-2H] DPMG 5 mM [U-2H] 'potassium phosphate' 20 mM 'natural abundance' 'sodium chloride' 50 mM 'natural abundance' EDTA 0.5 mM 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version . loop_ _Vendor _Address _Electronic_address Bruker . . stop_ loop_ _Task collection processing stop_ _Details . save_ save_SPARKY _Saveframe_category software _Name SPARKY _Version . loop_ _Vendor _Address _Electronic_address Goddard . . stop_ loop_ _Task 'chemical shift assignment' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model Avance _Field_strength 800 _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_3D_HNCO_2 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCO' _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_HNCACB_4 _Saveframe_category NMR_applied_experiment _Experiment_name '3D HNCACB' _Sample_label $sample_1 save_ save_3D_1H-15N_NOESY_5 _Saveframe_category NMR_applied_experiment _Experiment_name '3D 1H-15N NOESY' _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' 0.1 . M pH 7 . pH pressure 1 . atm temperature 318 . K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_DSS _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 0.00 internal direct . . . 0.251449530 DSS H 1 'methyl protons' ppm 0.00 internal direct . . . 1.000000000 DSS N 15 'methyl protons' ppm 0.00 internal direct . . . 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' '3D HNCO' '3D HNCA' '3D HNCACB' stop_ loop_ _Sample_label $sample_1 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $DSS _Mol_system_component_name BclxL _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 1 209 11 ARG H H 8.22 0.02 1 2 209 11 ARG C C 174.10 0.05 1 3 209 11 ARG CA C 54.72 0.05 1 4 209 11 ARG CB C 29.76 0.05 1 5 209 11 ARG N N 121.06 0.05 1 6 210 12 PHE H H 8.20 0.02 1 7 210 12 PHE C C 173.50 0.05 1 8 210 12 PHE CA C 56.08 0.05 1 9 210 12 PHE CB C 38.80 0.05 1 10 210 12 PHE N N 118.92 0.05 1 11 211 13 ASN H H 8.46 0.02 1 12 211 13 ASN C C 173.80 0.05 1 13 211 13 ASN CA C 52.21 0.05 1 14 211 13 ASN CB C 40.44 0.05 1 15 211 13 ASN N N 119.34 0.05 1 16 212 14 ARG H H 8.52 0.02 1 17 212 14 ARG C C 176.20 0.05 1 18 212 14 ARG CA C 54.72 0.05 1 19 212 14 ARG CB C 28.50 0.05 1 20 212 14 ARG N N 121.59 0.05 1 21 213 15 TRP H H 8.30 0.02 1 22 213 15 TRP HE1 H 10.47 0.02 1 23 213 15 TRP C C 175.70 0.05 1 24 213 15 TRP CA C 56.79 0.05 1 25 213 15 TRP CB C 31.13 0.05 1 26 213 15 TRP N N 121.79 0.05 1 27 213 15 TRP NE1 N 130.03 0.05 1 28 214 16 PHE H H 8.38 0.02 1 29 214 16 PHE C C 174.60 0.05 1 30 214 16 PHE CA C 58.15 0.05 1 31 214 16 PHE N N 121.01 0.05 1 32 215 17 LEU H H 8.33 0.02 1 33 215 17 LEU C C 177.60 0.05 1 34 215 17 LEU CA C 55.21 0.05 1 35 215 17 LEU N N 119.21 0.05 1 36 216 18 THR H H 8.34 0.02 1 37 216 18 THR C C 173.70 0.05 1 38 216 18 THR CA C 64.42 0.05 1 39 216 18 THR N N 118.33 0.05 1 40 217 19 GLY H H 8.75 0.02 1 41 217 19 GLY C C 172.20 0.05 1 42 217 19 GLY CA C 44.86 0.05 1 43 217 19 GLY N N 109.22 0.05 1 44 218 20 MET H H 8.52 0.02 1 45 218 20 MET C C 175.74 0.05 1 46 218 20 MET CA C 55.16 0.05 1 47 218 20 MET N N 119.20 0.05 1 48 219 21 THR H H 7.94 0.02 1 49 219 21 THR C C 173.50 0.05 1 50 219 21 THR CA C 64.86 0.05 1 51 219 21 THR N N 116.19 0.05 1 52 220 22 VAL H H 8.18 0.02 1 53 220 22 VAL C C 174.70 0.05 1 54 220 22 VAL CA C 64.15 0.05 1 55 220 22 VAL N N 120.42 0.05 1 56 221 23 ALA H H 8.32 0.02 1 57 221 23 ALA C C 176.60 0.05 1 58 221 23 ALA CA C 52.49 0.05 1 59 221 23 ALA CB C 17.27 0.05 1 60 221 23 ALA N N 119.80 0.05 1 61 222 24 GLY H H 8.53 0.02 1 62 222 24 GLY C C 172.10 0.05 1 63 222 24 GLY CA C 44.64 0.05 1 64 222 24 GLY N N 104.42 0.05 1 65 223 25 VAL H H 8.33 0.02 1 66 223 25 VAL C C 175.50 0.05 1 67 223 25 VAL CA C 64.04 0.05 1 68 223 25 VAL CB C 29.81 0.05 1 69 223 25 VAL N N 121.06 0.05 1 70 224 26 VAL H H 8.23 0.02 1 71 224 26 VAL C C 176.20 0.05 1 72 224 26 VAL CA C 63.66 0.05 1 73 224 26 VAL CB C 30.20 0.05 1 74 224 26 VAL N N 119.35 0.05 1 75 225 27 LEU H H 8.55 0.02 1 76 225 27 LEU C C 176.50 0.05 1 77 225 27 LEU CA C 55.37 0.05 1 78 225 27 LEU CB C 41.26 0.05 1 79 225 27 LEU N N 120.68 0.05 1 80 226 28 LEU H H 8.50 0.02 1 81 226 28 LEU C C 176.30 0.05 1 82 226 28 LEU CA C 55.16 0.05 1 83 226 28 LEU N N 118.96 0.05 1 84 227 29 GLY H H 8.44 0.02 1 85 227 29 GLY C C 173.10 0.05 1 86 227 29 GLY CA C 44.15 0.05 1 87 227 29 GLY N N 105.28 0.05 1 88 228 30 SER H H 8.03 0.02 1 89 228 30 SER C C 173.40 0.05 1 90 228 30 SER CA C 58.32 0.05 1 91 228 30 SER N N 116.32 0.05 1 92 229 31 LEU H H 7.88 0.02 1 93 229 31 LEU C C 175.30 0.05 1 94 229 31 LEU CA C 54.07 0.05 1 95 229 31 LEU N N 121.22 0.05 1 96 230 32 PHE H H 7.85 0.02 1 97 230 32 PHE C C 173.30 0.05 1 98 230 32 PHE CA C 55.16 0.05 1 99 230 32 PHE CB C 38.70 0.05 1 100 230 32 PHE N N 115.41 0.05 1 101 231 33 SER H H 7.74 0.02 1 102 231 33 SER C C 171.80 0.05 1 103 231 33 SER CA C 55.97 0.05 1 104 231 33 SER CB C 63.56 0.05 1 105 231 33 SER N N 114.68 0.05 1 106 232 34 ARG H H 8.13 0.02 1 107 232 34 ARG C C 172.90 0.05 1 108 232 34 ARG CA C 53.30 0.05 1 109 232 34 ARG CB C 29.76 0.05 1 110 232 34 ARG N N 123.27 0.05 1 111 233 35 LYS H H 7.93 0.02 1 112 233 35 LYS CA C 54.88 0.05 1 113 233 35 LYS CB C 32.72 0.05 1 114 233 35 LYS N N 127.98 0.05 1 stop_ save_