Heterogeneity of tethered agonist signaling in adhesion G protein-coupled receptors

Andrew N Dates; Daniel T D Jones; Jeffrey S Smith; Meredith A Skiba; Maria F Rich; Maggie M Burruss; Andrew C Kruse; Stephen C Blacklow

doi:10.1016/j.chembiol.2024.03.004

Heterogeneity of tethered agonist signaling in adhesion G protein-coupled receptors

Cell Chem Biol. 2024 Apr 4:S2451-9456(24)00120-X. doi: 10.1016/j.chembiol.2024.03.004. Online ahead of print.

Authors

Andrew N Dates¹, Daniel T D Jones¹, Jeffrey S Smith², Meredith A Skiba¹, Maria F Rich³, Maggie M Burruss¹, Andrew C Kruse¹, Stephen C Blacklow⁴

Affiliations

¹ Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115, USA.
² Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115, USA; Department of Dermatology, Brigham and Women's Hospital, 221 Longwood Avenue, Boston, MA 02115, USA.
³ University of Cincinnati School of Medicine, Department of Molecular Genetics, Biochemistry, and Microbiology, Cincinnati, OH 45267, USA.
⁴ Department of Biological Chemistry and Molecular Pharmacology, Blavatnik Institute, Harvard Medical School, Boston, MA 02115, USA; Department of Cancer Biology, Dana Farber Cancer Institute, Boston, MA 02215, USA. Electronic address: stephen_blacklow@hms.harvard.edu.

PMID: 38608683
DOI: 10.1016/j.chembiol.2024.03.004

Abstract

Adhesion G protein-coupled receptor (aGPCR) signaling influences development and homeostasis in a wide range of tissues. In the current model for aGPCR signaling, ligand binding liberates a conserved sequence that acts as an intramolecular, tethered agonist (TA), yet this model has not been evaluated systematically for all aGPCRs. Here, we assessed the TA-dependent activities of all 33 aGPCRs in a suite of transcriptional reporter, G protein activation, and β-arrestin recruitment assays using a new fusion protein platform. Strikingly, only ∼50% of aGPCRs exhibited robust TA-dependent activation, and unlike other GPCR families, aGPCRs showed a notable preference for G_12/13 signaling. AlphaFold2 predictions assessing TA engagement in the predicted intramolecular binding pocket aligned with the TA dependence of the cellular responses. This dataset provides a comprehensive resource to inform the investigation of all human aGPCRs and for targeting aGPCRs therapeutically.

Keywords: Adhesion GPCR; AlphaFold; BRET; G protein-coupled receptor; G12/13 signaling; aGPCR; activity profiling; family B2; tethered agonism; β-arrestin.