黄色荧光蛋白猝灭法分析气味受体活性。

Q4 Biochemistry, Genetics and Molecular Biology

Methods in molecular biology Pub Date : 2025-01-01 DOI:10.1007/978-1-0716-4466-9_11

JiWoo Choi, JaeHyung Koo

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引用次数: 0

摘要

气味受体（ORs）作为G蛋白偶联受体（GPCR）中最大的亚家族，被越来越多地认为是在鼻腔和各种鼻外生物过程中至关重要的膜蛋白。然而，由于配体的可用性有限，研究这些鼻外ORs的功能是具有挑战性的，这对全面的研究构成了重大障碍。大规模筛选体外试验，如卤素敏感黄色荧光蛋白（YFP）猝灭试验，是去孤儿ORs至关重要。该方案采用YFP猝灭法鉴定OR配体相互作用，从而推进OR去孤儿化研究。

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Yellow Fluorescent Protein Quenching Assay for Analyzing Odorant Receptor Activity.

Odorant receptors (ORs), recognized as the largest subfamily of G protein-coupled receptors (GPCR), are increasingly identified as membrane proteins crucial not only in nasal but also in various extra-nasal biological processes. However, researching the functions of these extra-nasal ORs is challenging due to the limited availability of ligands, posing a significant barrier to comprehensive studies. Large-scale screening with in vitro assays, such as the halide-sensitive yellow fluorescent protein (YFP) quenching assay, is crucial for deorphanizing ORs. This protocol employs the YFP quenching assay to identify OR-ligand interactions, thereby advancing OR deorphanization research.

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来源期刊

Methods in molecular biology Biochemistry, Genetics and Molecular Biology-Genetics

CiteScore

2.00

自引率

0.00%

发文量

3536

期刊介绍： For over 20 years, biological scientists have come to rely on the research protocols and methodologies in the critically acclaimed Methods in Molecular Biology series. The series was the first to introduce the step-by-step protocols approach that has become the standard in all biomedical protocol publishing. Each protocol is provided in readily-reproducible step-by-step fashion, opening with an introductory overview, a list of the materials and reagents needed to complete the experiment, and followed by a detailed procedure that is supported with a helpful notes section offering tips and tricks of the trade as well as troubleshooting advice.