Section of Receptor Biology & Signaling, Department of Physiology & Pharmacology, Karolinska Institutet, Stockholm, Sweden.
Institute of Pharmacology and Toxicology, University of Wuerzburg, Wuerzburg, Germany.
Methods Mol Biol. 2021;2268:137-147. doi: 10.1007/978-1-0716-1221-7_9.
Here we describe the stepwise application of bioluminescence resonance energy transfer (BRET)-based conformational receptor biosensors to study GPCR activation in intact cells. This technology can be easily adopted to various plate reader devices and microtiter plate formats. Due to the high sensitivity of these BRET-based receptor biosensors and their ability to quantify simultaneously receptor activation/de-activation kinetics as well as compound efficacy and potency, these optical tools provide the most direct and unbiased approach to monitor GPCR activity in a high-throughput-compatible assay format, representing a novel promising tool for the discovery of potential GPCR therapeutics.
在这里，我们描述了基于生物发光共振能量转移（BRET）的构象受体生物传感器在完整细胞中研究 G 蛋白偶联受体（GPCR）激活的逐步应用。该技术可以很容易地应用于各种平板读数器设备和微量滴定板格式。由于这些基于 BRET 的受体生物传感器具有高灵敏度，并且能够同时定量受体激活/失活动力学以及化合物的功效和效力，因此这些光学工具提供了最直接和最客观的方法来监测 GPCR 活性在高通量兼容的测定格式中，代表了一种用于发现潜在 GPCR 治疗药物的新型有前途的工具。
