Real-Time Monitoring of GPCR/cAMP Signalling by FRET and Single-Molecule Microscopy

摘要

G-protein-coupled receptors (GPCRs) are encoded by nearly 1000 genes in the human genome and mediate the effects of a variety of endogenous and exogenous cues, including, most importantly, several hormones and neurotrans-mitters. Because of their essential roles in physiology, their involvement in a plethora of diseases and their favourable characteristics as drug targets, they have been the focus of intensive investigation over more than 4 decades [1,2]. The properties of GPCRs have been studied long before their purification and the cloning of their genes using classical pharmacological assays, such as those measuring a physiological response in living animals or organ preparations and, later, with radioligand binding assays [2]. Thereafter, GPCRs and their signalling cascades have been mostly investigated by means of biochemical assays on crude cell preparations or reconstituted systems with purified proteins.