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Illuminating G-Protein-Coupling Selectivity of GPCRs

Inoue, Asuka, Raimondi, Francesco, Kadji, Francois Marie Ngako, Singh, Gurdeep, Kishi, Takayuki, Uwamizu, Akiharu, Ono, Yuki, Shinjo, Yuji, Ishida, Satoru, Arang, Nadia, Kawakami, Kouki, Gutkind, J. Silvio, Aoki, Junken, Russell, Robert B.
Cell 2019 v.177 no.7 pp. 1933-1947.e25
G-protein coupled receptors, G-proteins, data collection, dissociation, human cell lines, transforming growth factor alpha
Heterotrimetic G proteins consist of four subfamilies (Gs, Gi/o, Gq/11, and G12/13) that mediate signaling via G-protein-coupled receptors (GPCRs), principally by receptors binding Gα C termini. G-protein-coupling profiles govern GPCR-induced cellular responses, yet receptor sequence selectivity determinants remain elusive. Here, we systematically quantified ligand-induced interactions between 148 GPCRs and all 11 unique Gα subunit C termini. For each receptor, we probed chimeric Gα subunit activation via a transforming growth factor-α (TGF-α) shedding response in HEK293 cells lacking endogenous Gq/11 and G12/13 proteins, and complemented G-protein-coupling profiles through a NanoBiT-G-protein dissociation assay. Interrogation of the dataset identified sequence-based coupling specificity features, inside and outside the transmembrane domain, which we used to develop a coupling predictor that outperforms previous methods. We used the predictor to engineer designer GPCRs selectively coupled to G12. This dataset of fine-tuned signaling mechanisms for diverse GPCRs is a valuable resource for research in GPCR signaling.