biostudies-literatureMasuho IEuropean Research CouncilNIDA NIH HHSNIMH NIH HHS113173https://www.ebi.ac.uk/biostudies/studies/S-EPMC10842385biostudies-literatureUnknown42(10)G protein-coupled receptors (GPCRs) convert extracellular stimuli into intracellular signaling by coupling to heterotrimeric G proteins of four classes: G_i/o, G_q, G_s, and G_12/13. However, our understanding of the G protein selectivity of GPCRs is incomplete. Here, we quantitatively measure the enzymatic activity of GPCRs in living cells and reveal the G protein selectivity of 124 GPCRs with the exact rank order of their G protein preference. Using this information, we establish a classification of GPCRs by functional selectivity, discover the existence of a G_12/13-coupled receptor, G₁₅-coupled receptors, and a variety of subclasses for G_i/o-, G_q-, and G_s-coupled receptors, culminating in deveCell reportsRules and mechanisms governing G protein coupling selectivity of GPCRs.PMC10842385716058R01 DA036596R01 MH105482Li XMartemyanov KAMasuho IGainza PWakasugi-Masuho HKise RTany RCorreia BEVon Moo EfalseRules and mechanisms governing G protein coupling selectivity of GPCRs.G protein-coupled receptors (GPCRs) convert extracellular stimuli into intracellular signaling by coupling to heterotrimeric G proteins of four classes: G_i/o, G_q, G_s, and G_12/13. However, our understanding of the G protein selectivity of GPCRs is incomplete. Here, we quantitatively measure the enzymatic activity of GPCRs in living cells and reveal the G protein selectivity of 124 GPCRs with the exact rank order of their G protein preference. Using this information, we establish a classification of GPCRs by functional selectivity, discover the existence of a G_12/13-coupled receptor, G₁₅-coupled receptors, and a variety of subclasses for G_i/o-, G_q-, and G_s-coupled receptors, culminating in deve2023-01-01T00:00:00Z2023 Oct2026-04-30T21:25:20.124Z2025-04-20T03:00:12.991ZS-EPMC108423853774218910.1016/j.celrep.2023.113173