data_27328 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Chemical shifts of the human GPCR A2AAR in complex with the antagonist ZM241385 at pH 7 ; _BMRB_accession_number 27328 _BMRB_flat_file_name bmr27328.str _Entry_type original _Submission_date 2017-12-04 _Accession_date 2017-12-04 _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 Eddy Matthew T. . 2 Lee Ming-Yue . . 3 Gao Zhan-Guo . . 4 White Kate L. . 5 Didenko Tatiana . . 6 Horst Reto . . 7 Audet Martin . . 8 Stanczak Pawel . . 9 McClary Kyle M. . 10 Han 'Gye Won' . . 11 Jacobson Kenneth A. . 12 Stevens Raymond C. . 13 Wuthrich Kurt . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 14 "15N chemical shifts" 14 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2018-02-07 original BMRB . stop_ _Original_release_date 2017-12-05 save_ ############################# # Citation for this entry # ############################# save_Citation_1 _Saveframe_category entry_citation _Citation_full . _Citation_title ; Allosteric Coupling of Drug Binding and Intracellular Signaling in the A2A Adenosine Receptor ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 29290469 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Eddy Matthew T. . 2 Lee Ming-Yue . . 3 Gao Zhan-Guo . . 4 White Kate L. . 5 Didenko Tatiana . . 6 Horst Reto . . 7 Audet Martin . . 8 Stanczak Pawel . . 9 McClary Kyle M. . 10 Han 'Gye Won' . . 11 Jacobson Kenneth A. . 12 Stevens Raymond C. . 13 Wuthrich Kurt . . stop_ _Journal_abbreviation Cell _Journal_volume 172 _Journal_issue 1-2 _Journal_ISSN 1097-4172 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first 68 _Page_last 80 _Year 2018 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'Human A2A adenosine receptor in complex with ZM241385' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label 'Human A2AAR(1-316)' $Human_A2AAR stop_ _System_molecular_weight 37626.51 _System_physical_state native _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _Database_query_date . _Details 'ZM241385 is A2AAR antagonist.' save_ ######################## # Monomeric polymers # ######################## save_Human_A2AAR _Saveframe_category monomeric_polymer _Mol_type polymer _Mol_polymer_class protein _Name_common Human_A2AAR _Molecular_mass . _Mol_thiol_state 'not reported' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 335 _Mol_residue_sequence ; MDYKDDDDKMPIMGSSVYIT VELAIAVLAILGNVLVCWAV WLNSNLQNVTNYFVVSLAAA DIAVGVLAIPFAITISTGFC AACHGCLFIACFVLVLTQSS IFSLLAIAIDRYIAIRIPLR YNGLVTGTRAKGIIAICWVL SFAIGLTPMLGWNNCGQPKE GKQHSQGCGEGQVACLFEDV VPMNYMVYFNFFACVLVPLL LMLGVYLRIFLAARRQLKQM ESQPLPGERARSTLQKEVHA AKSLAIIVGLFALCWLPLHI INCFTFFCPDCSHAPLWLMY LAIVLSHTNSVVNPFIYAYR IREFRQTFRKIIRSHVLRQQ EPFKAHHHHHHHHHH ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 -8 MET 2 -7 ASP 3 -6 TYR 4 -5 LYS 5 -4 ASP 6 -3 ASP 7 -2 ASP 8 -1 ASP 9 0 LYS 10 1 MET 11 2 PRO 12 3 ILE 13 4 MET 14 5 GLY 15 6 SER 16 7 SER 17 8 VAL 18 9 TYR 19 10 ILE 20 11 THR 21 12 VAL 22 13 GLU 23 14 LEU 24 15 ALA 25 16 ILE 26 17 ALA 27 18 VAL 28 19 LEU 29 20 ALA 30 21 ILE 31 22 LEU 32 23 GLY 33 24 ASN 34 25 VAL 35 26 LEU 36 27 VAL 37 28 CYS 38 29 TRP 39 30 ALA 40 31 VAL 41 32 TRP 42 33 LEU 43 34 ASN 44 35 SER 45 36 ASN 46 37 LEU 47 38 GLN 48 39 ASN 49 40 VAL 50 41 THR 51 42 ASN 52 43 TYR 53 44 PHE 54 45 VAL 55 46 VAL 56 47 SER 57 48 LEU 58 49 ALA 59 50 ALA 60 51 ALA 61 52 ASP 62 53 ILE 63 54 ALA 64 55 VAL 65 56 GLY 66 57 VAL 67 58 LEU 68 59 ALA 69 60 ILE 70 61 PRO 71 62 PHE 72 63 ALA 73 64 ILE 74 65 THR 75 66 ILE 76 67 SER 77 68 THR 78 69 GLY 79 70 PHE 80 71 CYS 81 72 ALA 82 73 ALA 83 74 CYS 84 75 HIS 85 76 GLY 86 77 CYS 87 78 LEU 88 79 PHE 89 80 ILE 90 81 ALA 91 82 CYS 92 83 PHE 93 84 VAL 94 85 LEU 95 86 VAL 96 87 LEU 97 88 THR 98 89 GLN 99 90 SER 100 91 SER 101 92 ILE 102 93 PHE 103 94 SER 104 95 LEU 105 96 LEU 106 97 ALA 107 98 ILE 108 99 ALA 109 100 ILE 110 101 ASP 111 102 ARG 112 103 TYR 113 104 ILE 114 105 ALA 115 106 ILE 116 107 ARG 117 108 ILE 118 109 PRO 119 110 LEU 120 111 ARG 121 112 TYR 122 113 ASN 123 114 GLY 124 115 LEU 125 116 VAL 126 117 THR 127 118 GLY 128 119 THR 129 120 ARG 130 121 ALA 131 122 LYS 132 123 GLY 133 124 ILE 134 125 ILE 135 126 ALA 136 127 ILE 137 128 CYS 138 129 TRP 139 130 VAL 140 131 LEU 141 132 SER 142 133 PHE 143 134 ALA 144 135 ILE 145 136 GLY 146 137 LEU 147 138 THR 148 139 PRO 149 140 MET 150 141 LEU 151 142 GLY 152 143 TRP 153 144 ASN 154 145 ASN 155 146 CYS 156 147 GLY 157 148 GLN 158 149 PRO 159 150 LYS 160 151 GLU 161 152 GLY 162 153 LYS 163 154 GLN 164 155 HIS 165 156 SER 166 157 GLN 167 158 GLY 168 159 CYS 169 160 GLY 170 161 GLU 171 162 GLY 172 163 GLN 173 164 VAL 174 165 ALA 175 166 CYS 176 167 LEU 177 168 PHE 178 169 GLU 179 170 ASP 180 171 VAL 181 172 VAL 182 173 PRO 183 174 MET 184 175 ASN 185 176 TYR 186 177 MET 187 178 VAL 188 179 TYR 189 180 PHE 190 181 ASN 191 182 PHE 192 183 PHE 193 184 ALA 194 185 CYS 195 186 VAL 196 187 LEU 197 188 VAL 198 189 PRO 199 190 LEU 200 191 LEU 201 192 LEU 202 193 MET 203 194 LEU 204 195 GLY 205 196 VAL 206 197 TYR 207 198 LEU 208 199 ARG 209 200 ILE 210 201 PHE 211 202 LEU 212 203 ALA 213 204 ALA 214 205 ARG 215 206 ARG 216 207 GLN 217 208 LEU 218 209 LYS 219 210 GLN 220 211 MET 221 212 GLU 222 213 SER 223 214 GLN 224 215 PRO 225 216 LEU 226 217 PRO 227 218 GLY 228 219 GLU 229 220 ARG 230 221 ALA 231 222 ARG 232 223 SER 233 224 THR 234 225 LEU 235 226 GLN 236 227 LYS 237 228 GLU 238 229 VAL 239 230 HIS 240 231 ALA 241 232 ALA 242 233 LYS 243 234 SER 244 235 LEU 245 236 ALA 246 237 ILE 247 238 ILE 248 239 VAL 249 240 GLY 250 241 LEU 251 242 PHE 252 243 ALA 253 244 LEU 254 245 CYS 255 246 TRP 256 247 LEU 257 248 PRO 258 249 LEU 259 250 HIS 260 251 ILE 261 252 ILE 262 253 ASN 263 254 CYS 264 255 PHE 265 256 THR 266 257 PHE 267 258 PHE 268 259 CYS 269 260 PRO 270 261 ASP 271 262 CYS 272 263 SER 273 264 HIS 274 265 ALA 275 266 PRO 276 267 LEU 277 268 TRP 278 269 LEU 279 270 MET 280 271 TYR 281 272 LEU 282 273 ALA 283 274 ILE 284 275 VAL 285 276 LEU 286 277 SER 287 278 HIS 288 279 THR 289 280 ASN 290 281 SER 291 282 VAL 292 283 VAL 293 284 ASN 294 285 PRO 295 286 PHE 296 287 ILE 297 288 TYR 298 289 ALA 299 290 TYR 300 291 ARG 301 292 ILE 302 293 ARG 303 294 GLU 304 295 PHE 305 296 ARG 306 297 GLN 307 298 THR 308 299 PHE 309 300 ARG 310 301 LYS 311 302 ILE 312 303 ILE 313 304 ARG 314 305 SER 315 306 HIS 316 307 VAL 317 308 LEU 318 309 ARG 319 310 GLN 320 311 GLN 321 312 GLU 322 313 PRO 323 314 PHE 324 315 LYS 325 316 ALA 326 317 HIS 327 318 HIS 328 319 HIS 329 320 HIS 330 321 HIS 331 322 HIS 332 323 HIS 333 324 HIS 334 325 HIS 335 326 HIS 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 $Human_A2AAR 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 _Vector_name $Human_A2AAR 'recombinant technology' . Pichia pastoris Bg12 pPIC9k 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 $Human_A2AAR 250 uM '[U-15N; ~70% 2H]' ZM241385 1 mM 'natural abundance' 'HEPES pH 7' 25 mM 'natural abundance' LMNG/CHS 5 mM 'natural abundance' 'sodium chloride' 75 mM 'natural abundance' stop_ save_ ############################ # Computer software used # ############################ save_TOPSPIN _Saveframe_category software _Name TOPSPIN _Version 3.5pl2 loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . 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_TROSY_1 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N TROSY' _Sample_label $sample_1 save_ ####################### # Sample conditions # ####################### save_sample_conditions_1 _Saveframe_category sample_conditions _Details '25 mM HEPES pH 7, 75 mM NaCl, LMNG/CHS' loop_ _Variable_type _Variable_value _Variable_value_error _Variable_value_units pH 7 . 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 H 1 'methyl protons' ppm 0.00 internal direct . . . 1.000000000 DSS N 15 'methyl protons' ppm 0.00 na indirect . . . 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 TROSY' 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 'Human A2AAR(1-316)' _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 5 14 GLY H H 8.05 0.02 . 2 5 14 GLY N N 108.91 0.1 . 3 29 38 TRP HE1 H 10.16 0.02 . 4 29 38 TRP NE1 N 127.57 0.1 . 5 32 41 TRP HE1 H 9.96 0.02 . 6 32 41 TRP NE1 N 129.41 0.1 . 7 69 78 GLY H H 7.79 0.02 . 8 69 78 GLY N N 110.81 0.1 . 9 114 123 GLY H H 7.68 0.02 . 10 114 123 GLY N N 105.32 0.1 . 11 118 127 GLY H H 9.03 0.02 . 12 118 127 GLY N N 110.45 0.1 . 13 129 138 TRP HE1 H 9.79 0.02 . 14 129 138 TRP NE1 N 128.70 0.1 . 15 142 151 GLY H H 6.74 0.02 . 16 142 151 GLY N N 101.00 0.1 . 17 143 152 TRP HE1 H 9.95 0.02 . 18 143 152 TRP NE1 N 127.49 0.1 . 19 158 167 GLY H H 8.23 0.02 . 20 158 167 GLY N N 109.29 0.1 . 21 160 169 GLY H H 8.16 0.02 . 22 160 169 GLY N N 108.90 0.1 . 23 218 227 GLY H H 8.29 0.02 . 24 218 227 GLY N N 109.83 0.1 . 25 246 255 TRP HE1 H 8.42 0.02 . 26 246 255 TRP NE1 N 124.70 0.1 . 27 268 277 TRP HE1 H 10.42 0.02 . 28 268 277 TRP NE1 N 131.20 0.1 . stop_ save_