G蛋白偶联受体寡聚化检测技术——共振能量转移技术和蛋白质片段互补技术

Techniques of Measurement for G Protein-Coupled Receptor Oligomerization

蛋白质-蛋白质相互作用(protein-protein interactions,PPIs)在细胞内信号途径的分子机制中具有重要作用,文章叙述了活细胞中以非侵入性荧光和生物发光为基础检测特异性PPIs的方法如FRET、BRET、BiFC和BiLC,上述技术广泛应用于G蛋白偶联受体(G protein-coupled receptors,GPCRs)寡聚化的研究,单独或联合应用这些新技术是研究两个蛋白或多个蛋白PPIs的最佳方法,文章描述了上述生物技术的最新进展及其优缺点,着重阐述在PPIs方面的应用。

Spatiotemporal characterization of protein-protein interactions（PPIs） is essential in determining the molecular mechanism of intracellular signaling processes.In this review,new methodological strategies derived from non-invasive fluorescence and luminescence-based approaches（FRET,BRET,BiFC and BiLC） were discussed when applied to the study of G protein-coupled receptor（GPCR） oligomerization.These new mothods can be used to detect specific PPIs in live cells.These techniques alone or in combination with complementary methods can be extremely powerful approaches for PPI visualization,even between more than two proteins.Here a comprehensive update on all the biotechnological aspects,including the strength and weakness of new fluorescence and luminescence-based methodologies,with a specific focus on their application for studying PPIs was provided.