Zhou Yiwei, Meng Jiyong, Xu Chanjuan, Liu Jianfeng
Cellular Signaling Laboratory, Key Laboratory of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, China.
Bioland Laboratory, Guangzhou Regenerative Medicine and Health Guangdong Laboratory, Guangzhou, China.
Front Cell Dev Biol. 2021 May 10;9:611443. doi: 10.3389/fcell.2021.611443. eCollection 2021.
G protein-coupled receptors (GPCRs) represent one of the largest membrane protein families that participate in various physiological and pathological activities. Accumulating structural evidences have revealed how GPCR activation induces conformational changes to accommodate the downstream G protein or β-arrestin. Multiple GPCR functional assays have been developed based on Förster resonance energy transfer (FRET) and bioluminescence resonance energy transfer (BRET) sensors to monitor the conformational changes in GPCRs, GPCR/G proteins, or GPCR/β-arrestin, especially over the past two decades. Here, we will summarize how these sensors have been optimized to increase the sensitivity and compatibility for application in different GPCR classes using various labeling strategies, meanwhile provide multiple solutions in functional assays for high-throughput drug screening.
G蛋白偶联受体（GPCRs）是参与各种生理和病理活动的最大膜蛋白家族之一。越来越多的结构证据揭示了GPCR激活如何诱导构象变化以容纳下游G蛋白或β-抑制蛋白。基于荧光共振能量转移（FRET）和生物发光共振能量转移（BRET）传感器，已经开发了多种GPCR功能测定方法来监测GPCR、GPCR/G蛋白或GPCR/β-抑制蛋白的构象变化，尤其是在过去二十年中。在这里，我们将总结这些传感器如何通过各种标记策略进行优化，以提高在不同GPCR类别中的应用灵敏度和兼容性，同时为高通量药物筛选的功能测定提供多种解决方案。
