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Related Experiment Video

Updated: May 11, 2025

High-throughput Analysis of Mammalian Olfactory Receptors: Measurement of Receptor Activation via Luciferase Activity
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Yellow Fluorescent Protein Quenching Assay for Analyzing Odorant Receptor Activity.

JiWoo Choi1,2, JaeHyung Koo3,4,5

  • 1Department of New Biology, DGIST, Daegu, Republic of Korea.

Methods in Molecular Biology (Clifton, N.J.)
|April 18, 2025
PubMed
Summary

This study introduces a halide-sensitive yellow fluorescent protein (YFP) quenching assay protocol to identify odorant receptor (OR) ligands. This method aids in deorphanizing ORs and understanding their diverse biological functions.

Keywords:
Extra-nasal OR functionsFunctional assaysGPCRLigand screeningMembrane proteinsOR deorphanizationOdorant receptorOlfactory receptorSignal transductionYFP quenching assay

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Last Updated: May 11, 2025

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Pharmacology

Background:

  • Odorant receptors (ORs), the largest G protein-coupled receptor (GPCR) family, are vital membrane proteins involved in nasal and extra-nasal biological processes.
  • Research into extra-nasal OR functions is hindered by a scarcity of known ligands, complicating comprehensive studies.

Purpose of the Study:

  • To present a robust protocol utilizing the YFP quenching assay for identifying ligands that interact with ORs.
  • To facilitate the deorphanization of ORs and expand the understanding of their roles in various biological systems.

Main Methods:

  • The study employs a halide-sensitive yellow fluorescent protein (YFP) quenching assay for high-throughput screening.
  • This in vitro assay is designed to detect interactions between ORs and potential odorant ligands.

Main Results:

  • The YFP quenching assay protocol enables efficient identification of OR-ligand interactions.
  • Successful application of the assay advances the deorphanization of previously uncharacterized ORs.

Conclusions:

  • The YFP quenching assay is a critical tool for OR deorphanization research.
  • This protocol supports the investigation of OR functions beyond olfaction, particularly in extra-nasal biological processes.