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Illuminating G-Protein-Coupling Selectivity of GPCRs

Author:
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.
Source:
Cell 2019 v.177 no.7 pp. 1933-1947.e25
ISSN:
0092-8674
Subject:
G-protein coupled receptors, G-proteins, data collection, dissociation, human cell lines, transforming growth factor alpha
Abstract:
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.
Agid:
6466035