Choi JiWoo, Koo JaeHyung
Department of New Biology, DGIST, Daegu, Republic of Korea.
New Biology Research Center (NBRC), DGIST, Daegu, Republic of Korea.
Methods Mol Biol. 2025;2915:169-177. doi: 10.1007/978-1-0716-4466-9_11.
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.
嗅觉受体(ORs)被认为是G蛋白偶联受体(GPCR)中最大的亚家族,越来越多地被确定为不仅在鼻腔而且在各种鼻外生物学过程中起关键作用的膜蛋白。然而,由于配体的可用性有限,研究这些鼻外ORs的功能具有挑战性,这对全面研究构成了重大障碍。使用体外测定法进行大规模筛选,如卤化物敏感型黄色荧光蛋白(YFP)淬灭测定法,对于使ORs解孤儿至关重要。本方案采用YFP淬灭测定法来鉴定OR-配体相互作用,从而推进OR解孤儿研究。