NYMC Faculty Publications

Uncovering the Signalling, Structure and Function of the 20-HETE-GPR75 Pairing: Identifying the Chemokine CCL5 as a Negative Regulator of GPR75

Author Type(s)

Faculty

DOI

10.1111/bph.15525

Journal Title

British Journal of Pharmacology

First Page

3813

Last Page

3828

Document Type

Article

Publication Date

9-2021

Department

Pharmacology

Abstract

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.

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