data_27297 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Methyl-methionine resonance assignments for Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, bound to nanobody Nb6B9 and full agonist isoprenaline ; _BMRB_accession_number 27297 _BMRB_flat_file_name bmr27297.str _Entry_type original _Submission_date 2017-10-26 _Accession_date 2017-10-26 _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 Solt Andras . . 2 Bostock Mark J. . 3 Shrestha Binesh . . 4 Kumar Prashant . . 5 Warne Tony . . 6 Tate Christopher G. . 7 Nietlispach Daniel . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 2 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 14 "13C chemical shifts" 14 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2018-01-29 original BMRB . stop_ loop_ _Related_BMRB_accession_number _Relationship 27292 'Turkey beta-1 adrenergic receptor, Delta5 mutant, Apo form' 27293 'Turkey beta-1 adrenergic receptor, Delta5 mutant, Isoprenaline-bound form' 27294 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, Apo form' 27295 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, Isoprenaline-bound form' 27296 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, bound to nanobody Nb6B9, without orthosteric ligand' stop_ _Original_release_date 2017-10-26 save_ ############################# # Citation for this entry # ############################# save_entry_citation _Saveframe_category entry_citation _Citation_full . _Citation_title ; Insight into partial agonism by observing multiple equilibria for ligand-bound and Gs-mimetic nanobody-bound beta1-adrenergic receptor ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 29176642 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Solt Andras . . 2 Bostock Mark J. . 3 Shrestha Binesh . . 4 Kumar Prashant . . 5 Warne Tony . . 6 Tate Christopher G. . 7 Nietlispach Daniel . . stop_ _Journal_abbreviation 'Nat. Commun.' _Journal_name_full 'Nature communications' _Journal_volume 8 _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 1795 _Page_last 1795 _Year 2017 _Details . loop_ _Keyword GPCR 'Solution NMR' stop_ save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, bound to nanobody Nb6B9 and full agonist isoprenaline' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, conformer 1' $Turkey_B1AR_D5_L190M 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, conformer 2' $Turkey_B1AR_D5_L190M 'Nanobody Nb6B9' $Nb6B9 Isoprenaline $entity_5FW 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_Turkey_B1AR_D5_L190M _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Turkey_B1AR_D5_L190M _Molecular_mass . _Mol_thiol_state 'free and disulfide bound' loop_ _Biological_function 'G-protein coupled receptor' Gs-mimetic stop_ _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 317 _Mol_residue_sequence ; MGAELLSQQWEAGLSLLLAL VVLLIVAGNVLVIAAIGSTQ RLQTLTNLFITSLACADLVM GLLVVPFGATLVVRGTWLWG SFLCELWTSLDVLCVTASIW TLCVIAIDRYLAITSPFRYQ SLMTRARAKVIICTVWAISA LVSFLPIMLHWWRDEDPQAM KCYQDPGCCDFVTNRAYAIA SSIISFYIPLLIMIFVYLRV YREAKEQIRKIDRASKRKTS RVAAMREHKALKTLGIIMGV FTLCWLPFFLVNIVNVFNRD LVPDWLFVAFNWLGYANSAA NPIIYCRSPDFRKAFKRLLA FPRKADRRLHHHHHHHH ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 1 MET 2 2 GLY 3 33 ALA 4 34 GLU 5 35 LEU 6 36 LEU 7 37 SER 8 38 GLN 9 39 GLN 10 40 TRP 11 41 GLU 12 42 ALA 13 43 GLY 14 44 LEU 15 45 SER 16 46 LEU 17 47 LEU 18 48 LEU 19 49 ALA 20 50 LEU 21 51 VAL 22 52 VAL 23 53 LEU 24 54 LEU 25 55 ILE 26 56 VAL 27 57 ALA 28 58 GLY 29 59 ASN 30 60 VAL 31 61 LEU 32 62 VAL 33 63 ILE 34 64 ALA 35 65 ALA 36 66 ILE 37 67 GLY 38 68 SER 39 69 THR 40 70 GLN 41 71 ARG 42 72 LEU 43 73 GLN 44 74 THR 45 75 LEU 46 76 THR 47 77 ASN 48 78 LEU 49 79 PHE 50 80 ILE 51 81 THR 52 82 SER 53 83 LEU 54 84 ALA 55 85 CYS 56 86 ALA 57 87 ASP 58 88 LEU 59 89 VAL 60 90 MET 61 91 GLY 62 92 LEU 63 93 LEU 64 94 VAL 65 95 VAL 66 96 PRO 67 97 PHE 68 98 GLY 69 99 ALA 70 100 THR 71 101 LEU 72 102 VAL 73 103 VAL 74 104 ARG 75 105 GLY 76 106 THR 77 107 TRP 78 108 LEU 79 109 TRP 80 110 GLY 81 111 SER 82 112 PHE 83 113 LEU 84 114 CYS 85 115 GLU 86 116 LEU 87 117 TRP 88 118 THR 89 119 SER 90 120 LEU 91 121 ASP 92 122 VAL 93 123 LEU 94 124 CYS 95 125 VAL 96 126 THR 97 127 ALA 98 128 SER 99 129 ILE 100 130 TRP 101 131 THR 102 132 LEU 103 133 CYS 104 134 VAL 105 135 ILE 106 136 ALA 107 137 ILE 108 138 ASP 109 139 ARG 110 140 TYR 111 141 LEU 112 142 ALA 113 143 ILE 114 144 THR 115 145 SER 116 146 PRO 117 147 PHE 118 148 ARG 119 149 TYR 120 150 GLN 121 151 SER 122 152 LEU 123 153 MET 124 154 THR 125 155 ARG 126 156 ALA 127 157 ARG 128 158 ALA 129 159 LYS 130 160 VAL 131 161 ILE 132 162 ILE 133 163 CYS 134 164 THR 135 165 VAL 136 166 TRP 137 167 ALA 138 168 ILE 139 169 SER 140 170 ALA 141 171 LEU 142 172 VAL 143 173 SER 144 174 PHE 145 175 LEU 146 176 PRO 147 177 ILE 148 178 MET 149 179 LEU 150 180 HIS 151 181 TRP 152 182 TRP 153 183 ARG 154 184 ASP 155 185 GLU 156 186 ASP 157 187 PRO 158 188 GLN 159 189 ALA 160 190 MET 161 191 LYS 162 192 CYS 163 193 TYR 164 194 GLN 165 195 ASP 166 196 PRO 167 197 GLY 168 198 CYS 169 199 CYS 170 200 ASP 171 201 PHE 172 202 VAL 173 203 THR 174 204 ASN 175 205 ARG 176 206 ALA 177 207 TYR 178 208 ALA 179 209 ILE 180 210 ALA 181 211 SER 182 212 SER 183 213 ILE 184 214 ILE 185 215 SER 186 216 PHE 187 217 TYR 188 218 ILE 189 219 PRO 190 220 LEU 191 221 LEU 192 222 ILE 193 223 MET 194 224 ILE 195 225 PHE 196 226 VAL 197 227 TYR 198 228 LEU 199 229 ARG 200 230 VAL 201 231 TYR 202 232 ARG 203 233 GLU 204 234 ALA 205 235 LYS 206 236 GLU 207 237 GLN 208 238 ILE 209 239 ARG 210 240 LYS 211 241 ILE 212 242 ASP 213 243 ARG 214 272 ALA 215 273 SER 216 274 LYS 217 275 ARG 218 276 LYS 219 277 THR 220 278 SER 221 279 ARG 222 280 VAL 223 281 ALA 224 282 ALA 225 283 MET 226 284 ARG 227 285 GLU 228 286 HIS 229 287 LYS 230 288 ALA 231 289 LEU 232 290 LYS 233 291 THR 234 292 LEU 235 293 GLY 236 294 ILE 237 295 ILE 238 296 MET 239 297 GLY 240 298 VAL 241 299 PHE 242 300 THR 243 301 LEU 244 302 CYS 245 303 TRP 246 304 LEU 247 305 PRO 248 306 PHE 249 307 PHE 250 308 LEU 251 309 VAL 252 310 ASN 253 311 ILE 254 312 VAL 255 313 ASN 256 314 VAL 257 315 PHE 258 316 ASN 259 317 ARG 260 318 ASP 261 319 LEU 262 320 VAL 263 321 PRO 264 322 ASP 265 323 TRP 266 324 LEU 267 325 PHE 268 326 VAL 269 327 ALA 270 328 PHE 271 329 ASN 272 330 TRP 273 331 LEU 274 332 GLY 275 333 TYR 276 334 ALA 277 335 ASN 278 336 SER 279 337 ALA 280 338 ALA 281 339 ASN 282 340 PRO 283 341 ILE 284 342 ILE 285 343 TYR 286 344 CYS 287 345 ARG 288 346 SER 289 347 PRO 290 348 ASP 291 349 PHE 292 350 ARG 293 351 LYS 294 352 ALA 295 353 PHE 296 354 LYS 297 355 ARG 298 356 LEU 299 357 LEU 300 358 ALA 301 359 PHE 302 360 PRO 303 361 ARG 304 362 LYS 305 363 ALA 306 364 ASP 307 365 ARG 308 366 ARG 309 367 LEU 310 484 HIS 311 485 HIS 312 486 HIS 313 487 HIS 314 488 HIS 315 489 HIS 316 490 HIS 317 491 HIS stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ save_Nb6B9 _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Nb6B9 _Molecular_mass . _Mol_thiol_state 'free and disulfide bound' loop_ _Biological_function Gs-mimetic stop_ _Details . _Residue_count 132 _Mol_residue_sequence ; HHHHHHLVPRGSQVQLQESG GGLVQAGGSLRLSCAASGSI FALNIMGWYRQAPGKQRELV AAIHSGGTTNYANSVKGRFT ISRDNAANTVYLQMNSLKPE DTAVYYCNVKDFGAIIYDYD YWGQGTQVTVSS ; loop_ _Residue_seq_code _Residue_label 1 HIS 2 HIS 3 HIS 4 HIS 5 HIS 6 HIS 7 LEU 8 VAL 9 PRO 10 ARG 11 GLY 12 SER 13 GLN 14 VAL 15 GLN 16 LEU 17 GLN 18 GLU 19 SER 20 GLY 21 GLY 22 GLY 23 LEU 24 VAL 25 GLN 26 ALA 27 GLY 28 GLY 29 SER 30 LEU 31 ARG 32 LEU 33 SER 34 CYS 35 ALA 36 ALA 37 SER 38 GLY 39 SER 40 ILE 41 PHE 42 ALA 43 LEU 44 ASN 45 ILE 46 MET 47 GLY 48 TRP 49 TYR 50 ARG 51 GLN 52 ALA 53 PRO 54 GLY 55 LYS 56 GLN 57 ARG 58 GLU 59 LEU 60 VAL 61 ALA 62 ALA 63 ILE 64 HIS 65 SER 66 GLY 67 GLY 68 THR 69 THR 70 ASN 71 TYR 72 ALA 73 ASN 74 SER 75 VAL 76 LYS 77 GLY 78 ARG 79 PHE 80 THR 81 ILE 82 SER 83 ARG 84 ASP 85 ASN 86 ALA 87 ALA 88 ASN 89 THR 90 VAL 91 TYR 92 LEU 93 GLN 94 MET 95 ASN 96 SER 97 LEU 98 LYS 99 PRO 100 GLU 101 ASP 102 THR 103 ALA 104 VAL 105 TYR 106 TYR 107 CYS 108 ASN 109 VAL 110 LYS 111 ASP 112 PHE 113 GLY 114 ALA 115 ILE 116 ILE 117 TYR 118 ASP 119 TYR 120 ASP 121 TYR 122 TRP 123 GLY 124 GLN 125 GLY 126 THR 127 GLN 128 VAL 129 THR 130 VAL 131 SER 132 SER stop_ _Sequence_homology_query_date . _Sequence_homology_query_revised_last_date . save_ ############# # Ligands # ############# save_5FW _Saveframe_category ligand _Mol_type "non-polymer (NON-POLYMER)" _Name_common "entity_5FW (ISOPRENALINE)" _BMRB_code 5FW _PDB_code 5FW _Molecular_mass 211.258 _Mol_charge 0 _Mol_paramagnetic . _Mol_aromatic yes _Details . loop_ _Atom_name _PDB_atom_name _Atom_type _Atom_chirality _Atom_charge _Atom_oxidation_number _Atom_unpaired_electrons CAI CAI C . 0 . ? CAG CAG C . 0 . ? CAJ CAJ C . 0 . ? NAD NAD N . 0 . ? CAF CAF C . 0 . ? CAE CAE C . 0 . ? OAA OAA O . 0 . ? CAH CAH C . 0 . ? CAK CAK C . 0 . ? CAL CAL C . 0 . ? CAN CAN C . 0 . ? CAO CAO C . 0 . ? OAC OAC O . 0 . ? CAM CAM C . 0 . ? OAB OAB O . 0 . ? HAI1 HAI1 H . 0 . ? HAI2 HAI2 H . 0 . ? HAI3 HAI3 H . 0 . ? HAG HAG H . 0 . ? HAJ1 HAJ1 H . 0 . ? HAJ2 HAJ2 H . 0 . ? HAJ3 HAJ3 H . 0 . ? HAD HAD H . 0 . ? HAF1 HAF1 H . 0 . ? HAF2 HAF2 H . 0 . ? HAE HAE H . 0 . ? HAA HAA H . 0 . ? HAK HAK H . 0 . ? HAL HAL H . 0 . ? HAN HAN H . 0 . ? HAC HAC H . 0 . ? HAB HAB 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 CAI CAG ? ? SING CAG CAJ ? ? SING CAG NAD ? ? SING NAD CAF ? ? SING CAF CAE ? ? SING CAE OAA ? ? SING CAE CAH ? ? SING CAH CAK ? ? DOUB CAH CAL ? ? DOUB CAK CAM ? ? SING CAL CAN ? ? DOUB CAN CAO ? ? SING CAO OAC ? ? SING CAO CAM ? ? SING CAM OAB ? ? SING CAI HAI1 ? ? SING CAI HAI2 ? ? SING CAI HAI3 ? ? SING CAG HAG ? ? SING CAJ HAJ1 ? ? SING CAJ HAJ2 ? ? SING CAJ HAJ3 ? ? SING NAD HAD ? ? SING CAF HAF1 ? ? SING CAF HAF2 ? ? SING CAE HAE ? ? SING OAA HAA ? ? SING CAK HAK ? ? SING CAL HAL ? ? SING CAN HAN ? ? SING OAC HAC ? ? SING OAB HAB ? ? stop_ _Mol_thiol_state . _Sequence_homology_query_date . save_ #################### # Natural source # #################### save_natural_source _Saveframe_category natural_source loop_ _Mol_label _Organism_name_common _NCBI_taxonomy_ID _Superkingdom _Kingdom _Genus _Species $Turkey_B1AR_D5_L190M Turkey 9103 Eukaryota Metazoa Meleagris gallopavo $Nb6B9 Llama 9844 Eukaryota Metazoa Lama glama 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 $Turkey_B1AR_D5_L190M 'recombinant technology' . Spodoptera frugiperda . pBacPAK8 $Nb6B9 'recombinant technology' . Escherichia coli . pET26b(+) stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_L190M-Nb6B9-Iso _Saveframe_category sample _Sample_type solution _Details . loop_ _Mol_label _Concentration_value _Concentration_value_units _Concentration_min_value _Concentration_max_value _Isotopic_labeling $Turkey_B1AR_D5_L190M 90 uM 80 100 13Ce-Met $Nb6B9 350 uM 300 400 'natural abundance' $entity_5FW 1 mM . . 'natural abundance' NaCl 150 mM . . 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_CCPN_Analysis _Saveframe_category software _Name CCPN_Analysis _Version 2.4.2 loop_ _Vendor _Address _Electronic_address CCPN . . stop_ loop_ _Task 'chemical shift assignment' 'data analysis' 'peak picking' 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-13C_SOFAST-HMQC_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-13C SOFAST-HMQC' _Sample_label $L190M-Nb6B9-Iso 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 8.0 . pH pressure 1 . atm temperature 308 . 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 0.00 na indirect . . . 0.251449530 DSS H 1 'methyl protons' ppm 0.00 internal direct . . . 1.000000000 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_ _Software_label $CCPN_Analysis stop_ loop_ _Experiment_label '2D 1H-13C SOFAST-HMQC' stop_ loop_ _Sample_label $L190M-Nb6B9-Iso stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, conformer 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 1 1 MET HE H 2.022 0.025 1 2 1 1 MET CE C 17.076 0.025 1 3 153 123 MET HE H 2.123 0.025 1 4 153 123 MET CE C 16.255 0.025 1 5 178 148 MET HE H 1.972 0.025 1 6 178 148 MET CE C 17.367 0.025 1 7 190 160 MET HE H 1.552 0.025 1 8 190 160 MET CE C 16.548 0.025 1 9 223 193 MET HE H 1.431 0.025 1 10 223 193 MET CE C 17.044 0.025 1 11 283 225 MET HE H 2.030 0.025 1 12 283 225 MET CE C 17.329 0.025 1 13 296 238 MET HE H 2.019 0.025 1 14 296 238 MET CE C 16.465 0.025 1 stop_ save_ save_assigned_chemical_shifts_conformation_2 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Software_label $CCPN_Analysis stop_ loop_ _Experiment_label '2D 1H-13C SOFAST-HMQC' stop_ loop_ _Sample_label $L190M-Nb6B9-Iso stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chemical_shift_reference_1 _Mol_system_component_name 'Turkey beta-1 adrenergic receptor, Delta5-L190M mutant, conformer 2' _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 1 1 MET HE H 2.022 0.025 1 2 1 1 MET CE C 17.076 0.025 1 3 153 123 MET HE H 2.123 0.025 1 4 153 123 MET CE C 16.255 0.025 1 5 178 148 MET HE H 1.906 0.025 1 6 178 148 MET CE C 17.550 0.025 1 7 190 160 MET HE H 1.552 0.025 1 8 190 160 MET CE C 16.548 0.025 1 9 223 193 MET HE H 1.431 0.025 1 10 223 193 MET CE C 17.044 0.025 1 11 283 225 MET HE H 2.030 0.025 1 12 283 225 MET CE C 17.329 0.025 1 13 296 238 MET HE H 2.019 0.025 1 14 296 238 MET CE C 16.465 0.025 1 stop_ save_