title: Monitoring receptor signaling by intramolecular FRET.
abstract: A large variety of techniques has been used to monitor activation of G protein-coupled receptors (GPCRs) both in isolated membranes and in intact cells. However, most of these techniques cannot resolve receptor activation and signaling in space and in time. Here, we describe techniques that allow the temporally and spatially resolved monitoring of these processes by optical recording with energy transfer techniques. Fluorescence and bioluminescence resonance energy transfer, FRET and BRET, are based on energy transfer between two closely spaced probes. The exquisite sensitivity of FRET and BRET to the distance of the two probes makes these techniques ideal tools to study either protein-protein interactions (when the two probes are localized on two different proteins) or conformational changes within a given protein (when the two probes are localized on a single protein). Here, we review the latter approach as a tool to study receptor activation and the levels of the second messengers cAMP and cGMP in intact cells.
authors.affiliation: Institute of Pharmacology and Toxicology, University of Würzburg, Versbacher Str. 9, 97078 Würzburg, Germany. lohse@toxi.uni-wuerzburg.de
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Lohse
authors.forename: Moritz
authors.initials: M
authors.lastname: Bünemann
authors.forename: Carsten
authors.initials: C
authors.lastname: Hoffmann
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Vilardaga
authors.forename: Viacheslav O
authors.initials: VO
authors.lastname: Nikolaev
mesh_terms: D000818:Animals; D019390:Arrestins / Q000378:metabolism; D000242:Cyclic AMP / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D015398:Signal Transduction; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D043562:Receptors, G-Protein-Coupled; D000071557:beta-Arrestins; D000242:Cyclic AMP