data_36284 ####################### # Entry information # ####################### save_entry_information _Saveframe_category entry_information _Entry_title ; Structure of the beta2 adrenergic receptor in the full agonist bound state ; _BMRB_accession_number 36284 _BMRB_flat_file_name bmr36284.str _Entry_type original _Submission_date 2019-08-21 _Accession_date 2020-02-13 _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 Imai S. . . 2 Shimada I. . . stop_ loop_ _Saveframe_category_type _Saveframe_category_type_count assigned_chemical_shifts 1 stop_ loop_ _Data_type _Data_type_count "1H chemical shifts" 19 "15N chemical shifts" 19 stop_ loop_ _Revision_date _Revision_keyword _Revision_author _Revision_detail 2020-02-18 original BMRB . stop_ _Original_release_date 2020-02-13 save_ ############################# # Citation for this entry # ############################# save_citation_1 _Saveframe_category entry_citation _Citation_full . _Citation_title ; Structural equilibrium underlying ligand-dependent activation of beta2-adrenoreceptor ; _Citation_status published _Citation_type journal _CAS_abstract_code . _MEDLINE_UI_code . _PubMed_ID 31959965 loop_ _Author_ordinal _Author_family_name _Author_given_name _Author_middle_initials _Author_family_title 1 Imai S. . . 2 Yokomizo T. . . 3 Kofuku Y. . . 4 Shiraishi Y. . . 5 Ueda T. . . 6 Shimada I. . . stop_ _Journal_abbreviation 'Nat. Chem. Biol.' _Journal_volume . _Journal_issue . _Journal_ISSN 1552-4469 _Journal_CSD . _Book_chapter_title . _Book_volume . _Book_series . _Book_ISBN . _Conference_state_province . _Conference_abstract_number . _Page_first . _Page_last . _Year 2020 _Details . save_ ################################## # Molecular system description # ################################## save_assembly _Saveframe_category molecular_system _Mol_system_name 'beta 2 adrenergic receptor' _Enzyme_commission_number . loop_ _Mol_system_component_name _Mol_label entity_1 $entity_1 stop_ _System_molecular_weight . _System_oligomer_state ? _System_paramagnetic no _System_thiol_state . _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 38557.980 _Mol_thiol_state 'free and disulfide bound' _Details . ############################## # Polymer residue sequence # ############################## _Residue_count 336 _Mol_residue_sequence ; GTQQRDEVWVVGMGIVMSLI VLAIVFGNVLVITAICKFER LQTVTNYFITSLAVADLVMG LAVVPFGAAHILTKTWTFGN FWCEFWTSIDVLCVTASIWT LCVIAVDRYFAICSPFKYQS LLTKCKARVIILMVWIVSGL TSFLPIQMHWYRATHQEAIN CYAEETCCDFFTNQAYAIAS SIVSFYVPLVIMVFVYSRVF QEAKRQLQKIDKSEGRFHVQ NVSQVEQDGRTGHGHRRSSK FACKEHKALKTLGIIMGTFT LCWLPFFIVNIVHVIQDNLI RKEVYILLNWIGYVNSGFNP LIYSRSPDFRCAFQELLAAR RSSVKAHHHHHHHHHH ; loop_ _Residue_seq_code _Residue_author_seq_code _Residue_label 1 24 GLY 2 25 THR 3 26 GLN 4 27 GLN 5 28 ARG 6 29 ASP 7 30 GLU 8 31 VAL 9 32 TRP 10 33 VAL 11 34 VAL 12 35 GLY 13 36 MET 14 37 GLY 15 38 ILE 16 39 VAL 17 40 MET 18 41 SER 19 42 LEU 20 43 ILE 21 44 VAL 22 45 LEU 23 46 ALA 24 47 ILE 25 48 VAL 26 49 PHE 27 50 GLY 28 51 ASN 29 52 VAL 30 53 LEU 31 54 VAL 32 55 ILE 33 56 THR 34 57 ALA 35 58 ILE 36 59 CYS 37 60 LYS 38 61 PHE 39 62 GLU 40 63 ARG 41 64 LEU 42 65 GLN 43 66 THR 44 67 VAL 45 68 THR 46 69 ASN 47 70 TYR 48 71 PHE 49 72 ILE 50 73 THR 51 74 SER 52 75 LEU 53 76 ALA 54 77 VAL 55 78 ALA 56 79 ASP 57 80 LEU 58 81 VAL 59 82 MET 60 83 GLY 61 84 LEU 62 85 ALA 63 86 VAL 64 87 VAL 65 88 PRO 66 89 PHE 67 90 GLY 68 91 ALA 69 92 ALA 70 93 HIS 71 94 ILE 72 95 LEU 73 96 THR 74 97 LYS 75 98 THR 76 99 TRP 77 100 THR 78 101 PHE 79 102 GLY 80 103 ASN 81 104 PHE 82 105 TRP 83 106 CYS 84 107 GLU 85 108 PHE 86 109 TRP 87 110 THR 88 111 SER 89 112 ILE 90 113 ASP 91 114 VAL 92 115 LEU 93 116 CYS 94 117 VAL 95 118 THR 96 119 ALA 97 120 SER 98 121 ILE 99 122 TRP 100 123 THR 101 124 LEU 102 125 CYS 103 126 VAL 104 127 ILE 105 128 ALA 106 129 VAL 107 130 ASP 108 131 ARG 109 132 TYR 110 133 PHE 111 134 ALA 112 135 ILE 113 136 CYS 114 137 SER 115 138 PRO 116 139 PHE 117 140 LYS 118 141 TYR 119 142 GLN 120 143 SER 121 144 LEU 122 145 LEU 123 146 THR 124 147 LYS 125 148 CYS 126 149 LYS 127 150 ALA 128 151 ARG 129 152 VAL 130 153 ILE 131 154 ILE 132 155 LEU 133 156 MET 134 157 VAL 135 158 TRP 136 159 ILE 137 160 VAL 138 161 SER 139 162 GLY 140 163 LEU 141 164 THR 142 165 SER 143 166 PHE 144 167 LEU 145 168 PRO 146 169 ILE 147 170 GLN 148 171 MET 149 172 HIS 150 173 TRP 151 174 TYR 152 175 ARG 153 176 ALA 154 177 THR 155 178 HIS 156 179 GLN 157 180 GLU 158 181 ALA 159 182 ILE 160 183 ASN 161 184 CYS 162 185 TYR 163 186 ALA 164 187 GLU 165 188 GLU 166 189 THR 167 190 CYS 168 191 CYS 169 192 ASP 170 193 PHE 171 194 PHE 172 195 THR 173 196 ASN 174 197 GLN 175 198 ALA 176 199 TYR 177 200 ALA 178 201 ILE 179 202 ALA 180 203 SER 181 204 SER 182 205 ILE 183 206 VAL 184 207 SER 185 208 PHE 186 209 TYR 187 210 VAL 188 211 PRO 189 212 LEU 190 213 VAL 191 214 ILE 192 215 MET 193 216 VAL 194 217 PHE 195 218 VAL 196 219 TYR 197 220 SER 198 221 ARG 199 222 VAL 200 223 PHE 201 224 GLN 202 225 GLU 203 226 ALA 204 227 LYS 205 228 ARG 206 229 GLN 207 230 LEU 208 231 GLN 209 232 LYS 210 233 ILE 211 234 ASP 212 235 LYS 213 236 SER 214 237 GLU 215 238 GLY 216 239 ARG 217 240 PHE 218 241 HIS 219 242 VAL 220 243 GLN 221 244 ASN 222 245 VAL 223 246 SER 224 247 GLN 225 248 VAL 226 249 GLU 227 250 GLN 228 251 ASP 229 252 GLY 230 253 ARG 231 254 THR 232 255 GLY 233 256 HIS 234 257 GLY 235 258 HIS 236 259 ARG 237 260 ARG 238 261 SER 239 262 SER 240 263 LYS 241 264 PHE 242 265 ALA 243 266 CYS 244 267 LYS 245 268 GLU 246 269 HIS 247 270 LYS 248 271 ALA 249 272 LEU 250 273 LYS 251 274 THR 252 275 LEU 253 276 GLY 254 277 ILE 255 278 ILE 256 279 MET 257 280 GLY 258 281 THR 259 282 PHE 260 283 THR 261 284 LEU 262 285 CYS 263 286 TRP 264 287 LEU 265 288 PRO 266 289 PHE 267 290 PHE 268 291 ILE 269 292 VAL 270 293 ASN 271 294 ILE 272 295 VAL 273 296 HIS 274 297 VAL 275 298 ILE 276 299 GLN 277 300 ASP 278 301 ASN 279 302 LEU 280 303 ILE 281 304 ARG 282 305 LYS 283 306 GLU 284 307 VAL 285 308 TYR 286 309 ILE 287 310 LEU 288 311 LEU 289 312 ASN 290 313 TRP 291 314 ILE 292 315 GLY 293 316 TYR 294 317 VAL 295 318 ASN 296 319 SER 297 320 GLY 298 321 PHE 299 322 ASN 300 323 PRO 301 324 LEU 302 325 ILE 303 326 TYR 304 327 SER 305 328 ARG 306 329 SER 307 330 PRO 308 331 ASP 309 332 PHE 310 333 ARG 311 334 CYS 312 335 ALA 313 336 PHE 314 337 GLN 315 338 GLU 316 339 LEU 317 340 LEU 318 341 ALA 319 342 ALA 320 343 ARG 321 344 ARG 322 345 SER 323 346 SER 324 347 VAL 325 348 LYS 326 349 ALA 327 350 HIS 328 351 HIS 329 352 HIS 330 353 HIS 331 354 HIS 332 355 HIS 333 356 HIS 334 357 HIS 335 358 HIS 336 359 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 $entity_1 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 $entity_1 'recombinant technology' . Spodoptera frugiperda . . stop_ save_ ##################################### # Sample contents and methodology # ##################################### ######################## # Sample description # ######################## save_sample_1 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (A59C-MTSL), 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_10 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (I334C-MTSL), 1 mM ascorbate, 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' ascorbate 1 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_11 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (I334C-MTSL), 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_2 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (A59C-MTSL), 1 mM ascorbate, 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' ascorbate 1 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_3 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (T136C-MTSL), 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_4 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (T136C-MTSL), 1 mM ascorbate, 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' ascorbate 1 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_5 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (N148C-MTSL), 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_6 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (N148C-MTSL), 1 mM ascorbate, 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' ascorbate 1 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_7 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (L266C-MTSL), 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_8 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (L266C-MTSL), 1 mM ascorbate, 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' ascorbate 1 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ save_sample_9 _Saveframe_category sample _Sample_type micelle _Details '20 mM HEPES, 100 uM [2,3,3-2H,15N]-Leu beta 2 adrenergic receptor (I334C-MTSL), 95% H2O/5% D2O' loop_ _Mol_label _Concentration_value _Concentration_value_units _Isotopic_labeling HEPES 20 mM 'natural abundance' $entity_1 100 uM [2,3,3-2H,15N]-Leu H2O 95 % 'natural abundance' D2O 5 % [U-2H] stop_ save_ ############################ # Computer software used # ############################ save_software_1 _Saveframe_category software _Name CNS _Version . loop_ _Vendor _Address _Electronic_address 'Brunger A. T. et.al.' . . stop_ loop_ _Task refinement stop_ _Details . save_ save_software_2 _Saveframe_category software _Name TopSpin _Version . loop_ _Vendor _Address _Electronic_address 'Bruker Biospin' . . stop_ loop_ _Task 'chemical shift assignment' 'peak picking' stop_ _Details . save_ save_software_3 _Saveframe_category software _Name 'X-PLOR NIH' _Version . loop_ _Vendor _Address _Electronic_address 'Schwieters, Kuszewski, Tjandra and Clore' . . stop_ loop_ _Task 'structure calculation' stop_ _Details . save_ ######################### # Experimental detail # ######################### ################################## # NMR Spectrometer definitions # ################################## save_NMR_spectrometer_1 _Saveframe_category NMR_spectrometer _Manufacturer Bruker _Model 'AVANCE III' _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_2D_1H-15N_HSQC_2 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_2 save_ save_2D_1H-15N_HSQC_3 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_3 save_ save_2D_1H-15N_HSQC_4 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_4 save_ save_2D_1H-15N_HSQC_5 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_5 save_ save_2D_1H-15N_HSQC_6 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_6 save_ save_2D_1H-15N_HSQC_7 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_7 save_ save_2D_1H-15N_HSQC_8 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_8 save_ save_2D_1H-15N_HSQC_9 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_9 save_ save_2D_1H-15N_HSQC_10 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_10 save_ save_2D_1H-15N_HSQC_11 _Saveframe_category NMR_applied_experiment _Experiment_name '2D 1H-15N HSQC' _Sample_label $sample_11 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 . mM pH 6.5 0.1 pH pressure 1 . atm temperature 298 . K stop_ save_ #################### # NMR parameters # #################### ############################## # Assigned chemical shifts # ############################## ################################ # Chemical shift referencing # ################################ save_chem_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 0.000 external direct . . . 1.0 DSS N 15 'methyl protons' ppm 0.000 external 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_chemical_shifts_1 _Saveframe_category assigned_chemical_shifts _Details . loop_ _Experiment_label '2D 1H-15N HSQC' stop_ loop_ _Sample_label $sample_1 $sample_2 $sample_3 $sample_4 $sample_5 $sample_6 $sample_7 $sample_8 $sample_9 $sample_10 $sample_11 stop_ _Sample_conditions_label $sample_conditions_1 _Chem_shift_reference_set_label $chem_shift_reference_1 _Mol_system_component_name entity_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 19 LEU H H 7.96 0.02 1 2 42 19 LEU N N 122.09 0.30 1 3 45 22 LEU H H 8.23 0.02 1 4 45 22 LEU N N 117.04 0.30 1 5 53 30 LEU H H 8.01 0.02 1 6 53 30 LEU N N 123.43 0.30 1 7 64 41 LEU H H 7.88 0.02 1 8 64 41 LEU N N 117.45 0.30 1 9 80 57 LEU H H 8.90 0.02 1 10 80 57 LEU N N 122.03 0.30 1 11 84 61 LEU H H 8.13 0.02 1 12 84 61 LEU N N 114.37 0.30 1 13 115 92 LEU H H 8.64 0.02 1 14 115 92 LEU N N 124.09 0.30 1 15 144 121 LEU H H 8.04 0.02 1 16 144 121 LEU N N 120.20 0.30 1 17 145 122 LEU H H 7.61 0.02 1 18 145 122 LEU N N 120.86 0.30 1 19 155 132 LEU H H 8.21 0.02 1 20 155 132 LEU N N 120.15 0.30 1 21 163 140 LEU H H 8.65 0.02 1 22 163 140 LEU N N 122.57 0.30 1 23 167 144 LEU H H 7.15 0.02 1 24 167 144 LEU N N 113.94 0.30 1 25 212 189 LEU H H 7.95 0.02 1 26 212 189 LEU N N 116.72 0.30 1 27 230 207 LEU H H 7.85 0.02 1 28 230 207 LEU N N 120.52 0.30 1 29 266 243 CYS H H 8.11 0.02 1 30 266 243 CYS N N 121.21 0.30 1 31 284 261 LEU H H 8.39 0.02 1 32 284 261 LEU N N 124.84 0.30 1 33 287 264 LEU H H 7.81 0.02 1 34 287 264 LEU N N 120.47 0.30 1 35 311 288 LEU H H 8.31 0.02 1 36 311 288 LEU N N 114.41 0.30 1 37 340 317 LEU H H 7.77 0.02 1 38 340 317 LEU N N 116.41 0.30 1 stop_ save_