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GzESTY 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.
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.
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