Binkowski Brock F, Fan Frank, Wood Keith V
Promega Corporation, Madison, WI, USA.
Methods Mol Biol. 2011;756:263-71. doi: 10.1007/978-1-61779-160-4_14.
G-protein coupled, seven-transmembrane (7-TM) receptors (GPCRs) comprise a diverse class of signaling molecules involved in cellular physiology and pathology. In recent years, intracellular biosensors have been developed to monitor changes in intracellular cAMP in real time, facilitating studies on the mechanisms of GPCR activation and desensitization in living cells. However, methods based on fluorescence can show limitations in response dynamics together with being difficult to perform. Here we present the use of genetically encoded, luminescent biosensors that allow a facile, non-lytic assay format for monitoring cAMP dynamics in living cells.
G蛋白偶联的七跨膜（7-TM）受体（GPCRs）是一类参与细胞生理和病理过程的多样的信号分子。近年来，已经开发出细胞内生物传感器来实时监测细胞内cAMP的变化，有助于研究活细胞中GPCR激活和脱敏的机制。然而，基于荧光的方法在响应动力学方面可能存在局限性，并且操作困难。在这里，我们展示了使用基因编码的发光生物传感器，它允许以简便、非裂解的检测形式监测活细胞中的cAMP动态。
