Illuminating G-Protein-Coupling Selectivity of GPCRs

Overview

Journal Cell

Publisher Cell Press

Specialty Cell Biology

Date 2019 Jun 5

PMID 31160049

Citations 259

Authors

Asuka Inoue

Francesco Raimondi

Francois Marie Ngako Kadji

Gurdeep Singh

Takayuki Kishi

Akiharu Uwamizu

Yuki Ono

Yuji Shinjo

Satoru Ishida

Nadia Arang

Kouki Kawakami

J Silvio Gutkind

Junken Aoki

Robert B Russell

Affiliations

Soon will be listed here.

Abstract

Heterotrimetic G proteins consist of four subfamilies (G, G, G, and G) 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 G and G 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 G. This dataset of fine-tuned signaling mechanisms for diverse GPCRs is a valuable resource for research in GPCR signaling.

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