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| Title | Time-resolved cryo-EM of G-protein activation by a GPCR. |
|---|---|
| Journal, issue, pages | Nature, Year 2024 |
| Publish date | Mar 13, 2024 |
 Authors | Makaía M Papasergi-Scott / Guillermo Pérez-Hernández / Hossein Batebi / Yang Gao / Gözde Eskici / Alpay B Seven / Ouliana Panova / Daniel Hilger / Marina Casiraghi / Feng He / Luis Maul / Peter Gmeiner / Brian K Kobilka / Peter W Hildebrand / Georgios Skiniotis /    |
| PubMed Abstract | G-protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by stimulating guanine nucleotide exchange in the Gα subunit. To visualize this mechanism, we developed a time-resolved cryo-EM ...G-protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by stimulating guanine nucleotide exchange in the Gα subunit. To visualize this mechanism, we developed a time-resolved cryo-EM approach that examines the progression of ensembles of pre-steady-state intermediates of a GPCR-G-protein complex. By monitoring the transitions of the stimulatory G protein in complex with the β-adrenergic receptor at short sequential time points after GTP addition, we identified the conformational trajectory underlying G-protein activation and functional dissociation from the receptor. Twenty structures generated from sequential overlapping particle subsets along this trajectory, compared to control structures, provide a high-resolution description of the order of main events driving G-protein activation in response to GTP binding. Structural changes propagate from the nucleotide-binding pocket and extend through the GTPase domain, enacting alterations to Gα switch regions and the α5 helix that weaken the G-protein-receptor interface. Molecular dynamics simulations with late structures in the cryo-EM trajectory support that enhanced ordering of GTP on closure of the α-helical domain against the nucleotide-bound Ras-homology domain correlates with α5 helix destabilization and eventual dissociation of the G protein from the GPCR. These findings also highlight the potential of time-resolved cryo-EM as a tool for mechanistic dissection of GPCR signalling events. |
 External links | Nature / PubMed:38480881 |
| Methods | EM (single particle) |
| Resolution | 2.9 - 4.2 Å |
| Structure data | EMDB-29951, PDB-8gdz: EMDB-29952, PDB-8ge1: EMDB-29953, PDB-8ge2: EMDB-29954, PDB-8ge3: EMDB-29955, PDB-8ge4: EMDB-29956, PDB-8ge5: EMDB-29958, PDB-8ge6: EMDB-29959, PDB-8ge7: EMDB-29960, PDB-8ge8: EMDB-29961, PDB-8ge9: EMDB-29962, PDB-8gea: EMDB-29964, PDB-8geb: EMDB-29965, PDB-8gec: EMDB-29966, PDB-8ged: EMDB-29967, PDB-8gee: EMDB-29968, PDB-8gef: EMDB-29969, PDB-8geg: EMDB-29970, PDB-8geh: EMDB-29971, PDB-8gei: EMDB-29972, PDB-8gej: EMDB-29985, PDB-8gfv: EMDB-29986, PDB-8gfw: EMDB-29987, PDB-8gfx: EMDB-29988, PDB-8gfy: EMDB-29989, PDB-8gfz: EMDB-29990, PDB-8gg0: EMDB-29991, PDB-8gg1: EMDB-29992, PDB-8gg2: EMDB-29993, PDB-8gg3: EMDB-29994, PDB-8gg4: EMDB-29995, PDB-8gg5: EMDB-29996, PDB-8gg6: EMDB-29997, PDB-8gg7: EMDB-29998, PDB-8gg8: EMDB-29999, PDB-8gg9: EMDB-40000, PDB-8gga: EMDB-40001, PDB-8ggb: EMDB-40002, PDB-8ggc: EMDB-40004, PDB-8gge: EMDB-40005, PDB-8ggf: EMDB-40009, PDB-8ggi: EMDB-40010, PDB-8ggj: EMDB-40011, PDB-8ggk: EMDB-40012, PDB-8ggl: EMDB-40013, PDB-8ggm: EMDB-40014, PDB-8ggn: EMDB-40015, PDB-8ggo: EMDB-40016, PDB-8ggp: EMDB-40017, PDB-8ggq: EMDB-40018, PDB-8ggr: EMDB-40019, PDB-8ggs: EMDB-40020, PDB-8ggt: EMDB-40021, PDB-8ggu: EMDB-40022, PDB-8ggv: EMDB-40023, PDB-8ggw: EMDB-40024, PDB-8ggx: EMDB-40025, PDB-8ggy: EMDB-40026, PDB-8ggz: EMDB-40027, PDB-8gh0: EMDB-40028, PDB-8gh1:  EMDB-40096: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#1 of 20)  EMDB-40097: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#2 of 20)  EMDB-40098: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#3 of 20)  EMDB-40099: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#4 of 20)  EMDB-40100: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#5 of 20)  EMDB-40101: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#6 of 20)  EMDB-40102: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#7 of 20)  EMDB-40103: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#8 of 20)  EMDB-40104: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#9 of 20)  EMDB-40105: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#10 of 20)  EMDB-40106: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#11 of 20)  EMDB-40107: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#12 of 20)  EMDB-40108: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#13 of 20)  EMDB-40109: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#14 of 20)  EMDB-40110: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#15 of 20)  EMDB-40111: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#16 of 20)  EMDB-40112: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#17 of 20)  EMDB-40113: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#18 of 20)  EMDB-40114: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#19 of 20)  EMDB-40115: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 5 sec after GTP addition (#20 of 20)  EMDB-40116: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#1 of 20)  EMDB-40117: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#2 of 20)  EMDB-40118: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#3 of 20)  EMDB-40119: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#4 of 20)  EMDB-40120: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#5 of 20)  EMDB-40121: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#6 of 20)  EMDB-40122: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#7 of 20)  EMDB-40123: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#8 of 20)  EMDB-40124: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#9 of 20)  EMDB-40125: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#10 of 20)  EMDB-40126: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#11 of 20)  EMDB-40127: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#12 of 20)  EMDB-40128: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#13 of 20)  EMDB-40129: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#14 of 20)  EMDB-40130: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#15 of 20)  EMDB-40131: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#16 of 20)  EMDB-40132: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#17 of 20)  EMDB-40133: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#18 of 20)  EMDB-40134: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#19 of 20)  EMDB-40135: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 10 sec after GTP addition (#20 of 20)  EMDB-40136: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#1 of 20)  EMDB-40137: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#2 of 20)  EMDB-40138: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#3 of 20)  EMDB-40139: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#4 of 20)  EMDB-40140: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#5 of 20)  EMDB-40141: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#6 of 20)  EMDB-40142: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#7 of 20)  EMDB-40143: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#8 of 20)  EMDB-40144: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#9 of 20)  EMDB-40145: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#10 of 20)  EMDB-40146: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#11 of 20)  EMDB-40147: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#12 of 20)  EMDB-40148: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#13 of 20)  EMDB-40149: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#14 of 20)  EMDB-40150: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#15 of 20)  EMDB-40151: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#16 of 20)  EMDB-40152: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#17 of 20)  EMDB-40153: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#18 of 20)  EMDB-40154: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#19 of 20)  EMDB-40155: CryoEM structure of beta-2-adrenergic receptor in complex with Gs heterotrimer, 17 sec after GTP addition (#20 of 20)  EMDB-40156: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #1 of 20)  EMDB-40157: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #2 of 20)  EMDB-40158: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #3 of 20)  EMDB-40159: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #4 of 20)  EMDB-40160: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #5 of 20)  EMDB-40161: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #6 of 20)  EMDB-40163: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #7 of 20)  EMDB-40164: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #8 of 20)  EMDB-40165: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #9 of 20)  EMDB-40166: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #10 of 20)  EMDB-40167: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #11 of 20)  EMDB-40168: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #12 of 20)  EMDB-40169: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #13 of 20)  EMDB-40170: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #14 of 20)  EMDB-40171: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #15 of 20)  EMDB-40172: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #16 of 20)  EMDB-40173: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #17 of 20)  EMDB-40174: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #18 of 20)  EMDB-40175: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #19 of 20)  EMDB-40176: Locally refined cryoEM structure of G protein heterotrimer from beta-2-adrenergic receptor in complex with GTP-bound Gs heterotrimer (transition intermediate #20 of 20) EMDB-42408, PDB-8unl: EMDB-42409, PDB-8unm: EMDB-42410, PDB-8unn: EMDB-42411, PDB-8uno: EMDB-42412, PDB-8unp: EMDB-42413, PDB-8unq: EMDB-42414, PDB-8unr: EMDB-42415, PDB-8uns: EMDB-42416, PDB-8unt: EMDB-42417, PDB-8unu: EMDB-42418, PDB-8unv: EMDB-42419, PDB-8unw: EMDB-42420, PDB-8unx: EMDB-42421, PDB-8uny: EMDB-42422, PDB-8unz: EMDB-42423, PDB-8uo0: EMDB-42424, PDB-8uo1: EMDB-42425, PDB-8uo2: EMDB-42426, PDB-8uo3: EMDB-42427, PDB-8uo4: |
| Chemicals |  ChemComp-G1I:  ChemComp-HOH:  ChemComp-GTP:  ChemComp-MG: |
| Source |
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 Keywords |  SIGNALING PROTEIN / GPCR / Adrenergic / Receptor / G protein |
