Minute-scale persistence of a GPCR conformation state triggered by non-cognate G protein interactions primes signaling

Tejas M. Gupte, Michael Ritt, Matthew Dysthe, Rabia U. Malik, Sivaraj Sivaramakrishnan

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Despite the crowded nature of the cellular milieu, ligand–GPCR–G protein interactions are traditionally viewed as spatially and temporally isolated events. In contrast, recent reports suggest the spatial and temporal coupling of receptor–effector interactions, with the potential to diversify downstream responses. In this study, we combine protein engineering of GPCR–G protein interactions with affinity sequestration and photo-manipulation of the crucial Gα C terminus, to demonstrate the temporal coupling of cognate and non-cognate G protein interactions through priming of the GPCR conformation. We find that interactions of the Gαs and Gαq C termini with the β2-adrenergic receptor (β2-AR), targeted at the G-protein-binding site, enhance Gs activation and cyclic AMP levels. β2-AR–Gα C termini interactions alter receptor conformation, which persists for ~90 s following Gα C terminus dissociation. Non-cognate G-protein expression levels impact cognate signaling in cells. Our study demonstrates temporal allostery in GPCRs, with implications for the modulation of downstream responses through the canonical G-protein-binding interface.

Original languageEnglish (US)
Article number4836
JournalNature communications
Volume10
Issue number1
DOIs
StatePublished - Dec 1 2019

Bibliographical note

Funding Information:
We thank Todd Markowski of the Center of Mass Spectrometry and Proteomics, University of Minnesota College of Biological Sciences, for assistance with mass spectrometry experiments and analysis. Research was funded by the NIH (1R35 GM126940–01 to S.S.). We would like to thank Nurani Krishnan Sivaramakrishnan for his enduring support.

Publisher Copyright:
© 2019, The Author(s).

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