Uncovering the Signalling, Structure and Function of the 20-HETE-GPR75 Pairing: Identifying the Chemokine CCL5 as a Negative Regulator of GPR75
BACKGROUND AND PURPOSE: The G-protein-coupled receptor GPR75 (Gq) and its ligand, the cytochrome P450-derived vasoactive eicosanoid 20-hydroxyeicosatetraenoic acid (20-HETE), are involved in the activation of pro-inflammatory and hypertensive signalling cascades contributing to diabetes, obesity, vascular dysfunction/remodelling, hypertension and cardiovascular disease. Little is known as to how, where and with what affinity 20-HETE interacts with GPR75.
EXPERIMENTAL APPROACH: To better understand the pairing of 20-HETE and its receptor (GPR75), we used surface plasmon resonance (SPR) to determine binding affinity/kinetics. The PRESTO-Tango receptor-ome methodology for GPR75 overexpression was coupled with FLIPR Calcium 6 assays, homogeneous time-resolved fluorescence (HTRF) IP-1 and β-arrestin recruitment assays to determine receptor activation and downstream signalling events.
KEY RESULTS: SPR confirmed 20-HETE binding to GPR75 with an estimated K
CONCLUSIONS AND IMPLICATIONS: We have identified 20-HETE as a high-affinity ligand for GPR75 and CCL5 as a low-affinity negative regulator of GPR75, providing additional evidence for the deorphanization of GPR75 as a 20-HETE receptor.
Pascale, J. V., Park, E., Adebesin, A., Falck, J. R., Laniado Schwartzman, M., & Garcia, V. (2021). Uncovering the Signalling, Structure and Function of the 20-HETE-GPR75 Pairing: Identifying the Chemokine CCL5 as a Negative Regulator of GPR75. British Journal of Pharmacology, 178 (18), 3813-3828. https://doi.org/10.1111/bph.15525