Time-resolved cryo-EM of G-protein activation by a GPCR

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 and Georgios Skiniotis ()
Additional contact information
Makaía M. Papasergi-Scott: Stanford University School of Medicine
Guillermo Pérez-Hernández: Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Institute of Medical Physics and Biophysics
Hossein Batebi: Leipzig University
Yang Gao: Stanford University School of Medicine
Gözde Eskici: Stanford University School of Medicine
Alpay B. Seven: Stanford University School of Medicine
Ouliana Panova: Stanford University School of Medicine
Daniel Hilger: Stanford University School of Medicine
Marina Casiraghi: Stanford University School of Medicine
Feng He: Stanford University School of Medicine
Luis Maul: Friedrich-Alexander University Erlangen-Nürnberg
Peter Gmeiner: Friedrich-Alexander University Erlangen-Nürnberg
Brian K. Kobilka: Stanford University School of Medicine
Peter W. Hildebrand: Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin and Humboldt-Universität zu Berlin, Institute of Medical Physics and Biophysics
Georgios Skiniotis: Stanford University School of Medicine

Nature, 2024, vol. 629, issue 8014, 1182-1191

Abstract: Abstract G-protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by stimulating guanine nucleotide exchange in the Gα subunit1. 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 Gs protein in complex with the β2-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.

Date: 2024
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41586-024-07153-1 Abstract (text/html)
Access to the full text of the articles in this series is restricted.

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:nature:v:629:y:2024:i:8014:d:10.1038_s41586-024-07153-1

Ordering information: This journal article can be ordered from
https://www.nature.com/

DOI: 10.1038/s41586-024-07153-1

Access Statistics for this article

Nature is currently edited by Magdalena Skipper

More articles in Nature from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().