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Related Experiment Video

Updated: May 19, 2025

High-Speed Magnetic Tweezers for Nanomechanical Measurements on Force-Sensitive Elements

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Published on: May 12, 2023

G_zESTY as an optimized cell-based assay for initial steps in GPCR deorphanization.

Luca Franchini¹, Joseph J Porter¹, John D Lueck¹

¹Department of Pharmacology and Physiology, University of Rochester Medical Center, Rochester, NY, 14642, USA.

Nature Communications

|May 15, 2025

Summary

A new assay, GzESTY, aids in discovering ligands for G protein-coupled receptors (GPCRs), particularly orphan GPCRs. This tool helps identify unknown drug targets and understand GPCR functions.

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Area of Science:

Pharmacology
Molecular Biology
Assay Development

Background:

G protein-coupled receptors (GPCRs) are crucial drug targets, but many, especially orphan GPCRs, are underutilized due to unknown ligands and activation mechanisms.
Most GPCRs signal through the G$_{i/o/z}$ family, yet current activation assays are limited, impeding ligand discovery.

Purpose of the Study:

To introduce GzESTY, a novel, sensitive cell-based assay for studying G$_{i/o/z}$-coupled GPCR pharmacology.
To facilitate the deorphanization of orphan GPCRs by enabling ligand identification.

Main Methods:

Development and optimization of the GzESTY assay using an all-in-one vector for precise component expression.
Screening a library of ligand-activated GPCRs and endogenous GPCRs to assess assay performance.

Main Results:

GzESTY successfully detected activation of various GPCRs, differentiating between full and partial agonism.
The assay identified endogenous ligands for GPR176 and GPR37 in brain extracts, demonstrating its deorphanization utility.

Conclusions:

GzESTY is a sensitive and versatile tool for GPCR pharmacology research and ligand discovery.
This assay expands the capabilities for identifying ligands of orphan GPCRs, advancing drug target exploration.