Ferrer Marc, Kolodin Garrett D, Zuck Paul, Peltier Richard, Berry Kurtis, Mandala Suzanne M, Rosen Hugh, Ota Hisashi, Ozaki Satoshi, Inglese James, Strulovici Berta
Department of Automated Biotechnology, Merck Research Laboratories, North Wales, Pennsylvania, USA.
Assay Drug Dev Technol. 2003 Apr;1(2):261-73. doi: 10.1089/15406580360545071.
The diversity of physiological functions mediated by the GPCR superfamily provides a rich source of molecular targets for drug discovery programs. Consequently, a variety of assays have been designed to identify lead molecules based on ligand binding or receptor function. In one of these, the binding of [(35)S]GTPgammaS, a nonhydrolyzable analogue of GTP, to receptor-activated G-protein alpha subunits represents a unique functional assay for GPCRs and is well suited for use with automated HTS. Here we compare [(35)S]GTPgammaS scintillation proximity binding assays for two different G(i)-coupled GPCRs, and describe their implementation with automated high-throughput systems.
G蛋白偶联受体（GPCR）超家族介导的生理功能多样性为药物研发项目提供了丰富的分子靶点来源。因此，人们设计了各种检测方法，以基于配体结合或受体功能来鉴定先导分子。其中一种方法是，[(35)S]GTPγS（一种GTP的不可水解类似物）与受体激活的G蛋白α亚基结合，这是一种针对GPCR的独特功能检测方法，非常适合用于自动化的高通量筛选（HTS）。在这里，我们比较了针对两种不同的G(i)偶联GPCR的[(35)S]GTPγS闪烁邻近结合检测方法，并描述了它们在自动化高通量系统中的应用。
