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Spatiotemporally resolved GPCR interactome uncovers unique mediators of receptor agonism.

Maria M Shchepinova·Rachel Richardson·Jack W Houghton·Abigail R Walker·Mohammed A Safar·Daniel Conole·Aylin C Hanyaloglu·Edward W Tate
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Abstract

Cellular signaling by membrane G protein-coupled receptors (GPCRs) is governed by a complex and diverse array of mechanisms. The dynamics of a GPCR interactome, as it evolves over time and space in response to an agonist, provide a unique perspective on pleiotropic signaling decoding and functional selectivity at the cellular level. In this study, we utilized proximity-based APEX2 proteomics to investigate the interaction network of the luteinizing hormone receptor (LHR) on a minute-to-minute timescale. We developed an analytical approach that integrates quantitative multiplexed proteomics with temporal reference profiles, creating a platform to identify the proteomic environment of APEX2-tagged LHR at the nanometer scale. LHR activity is finely regulated spatially, leading to the identification of putative interactors, including the Ras-related GTPase RAP2B, which modulate both receptor signaling and post-endocytic trafficking. This work provides a valuable resource for spatiotemporal nanodomain mapping of LHR interactors across subcellular compartments.

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HumansHEK293 CellsReceptors, LHProteomicsSignal TransductionReceptors, G-Protein-CoupledAPEX2G protein-coupled receptorGPCRLHRendocytosisendosomeinteractomeisobaric taggingluteinizing hormone receptormass spectrometryproximity labelingproximity proteomicssignalingvery early endosomeJournal ArticleResearch Support, Non-U.S. Gov't

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Maria M Shchepinova, Rachel Richardson, Jack W Houghton, Abigail R Walker, Mohammed A Safar, Daniel Conole, Aylin C Hanyaloglu, Edward W Tate. 2025-05-15. Spatiotemporally resolved GPCR interactome uncovers unique mediators of receptor agonism.. https://doi.org/10.1016/j.chembiol.2025.04.006

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