Unveiling Mechanical Activation: GAIN Domain Unfolding and Dissociation in Adhesion GPCRs

Chaoyu Fu; Wenmao Huang; Qingnan Tang; Minghui Niu; Shiwen Guo; Tobias Langenhan; Gaojie Song; Jie Yan

doi:10.1021/acs.nanolett.3c01163

Unveiling Mechanical Activation: GAIN Domain Unfolding and Dissociation in Adhesion GPCRs

Nano Lett. 2023 Oct 25;23(20):9179-9186. doi: 10.1021/acs.nanolett.3c01163. Epub 2023 Oct 13.

Authors

Chaoyu Fu^{1

2}, Wenmao Huang^{1

2}, Qingnan Tang¹, Minghui Niu³, Shiwen Guo², Tobias Langenhan⁴, Gaojie Song³, Jie Yan^{1

2

5

6}

Affiliations

¹ Department of Physics, National University of Singapore, Singapore 117551, Singapore.
² Mechanobiology Institute, National University of Singapore, Singapore 117411, Singapore.
³ School of Life Sciences, East China Normal University, Shanghai 200241, China.
⁴ Rudolf Schönheimer Institute of Biochemistry, Division of General Biochemistry, Medical Faculty, Leipzig University, Leipzig 04103, Germany.
⁵ Centre for Bioimaging Sciences, National University of Singapore, Singapore 117557, Singapore.
⁶ Joint School of National University of Singapore and Tianjin University, International Campus of Tianjin University, Binhai New City, Fuzhou 350207, China.

PMID: 37831892
PMCID: PMC10607210 (available on 2024-10-13)
DOI: 10.1021/acs.nanolett.3c01163

Abstract

Adhesion G protein-coupled receptors (aGPCRs) have extracellular regions (ECRs) containing GPCR autoproteolysis-inducing (GAIN) domains. The GAIN domain enables the ECR to self-cleave into N- and C-terminal fragments. However, the impact of force on the GAIN domain's conformation, critical for mechanosensitive aGPCR activation, remains unclear. Our study investigated the mechanical stability of GAIN domains in three aGPCRs (B, G, and L subfamilies) at a loading rate of 1 pN/s. We discovered that forces of a few piconewtons can destabilize the GAIN domains. In autocleaved aGPCRs ADGRG1/GPR56 and ADGRL1/LPHN1, these forces cause the GAIN domain detachment from the membrane-proximal Stachel sequence, preceded by partial unfolding. In noncleavable aGPCR ADGRB3/BAI3 and cleavage-deficient mutant ADGRG1/GPR56-T383G, complex mechanical unfolding of the GAIN domain occurs. Additionally, GAIN domain detachment happens during cell migration. Our findings support the mechanical activation hypothesis of aGPCRs, emphasizing the sensitivity of the GAIN domain structure and detachment to physiological force ranges.

Keywords: BAI3; GAIN domain dissociation; GAIN domain unfolding; GPR56; LPHN1; adhesion GPCR.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cell Adhesion
Models, Molecular
Receptors, G-Protein-Coupled* / chemistry
Receptors, G-Protein-Coupled* / genetics

Substances

Receptors, G-Protein-Coupled