title: Kinetic analysis of G protein-coupled receptor signaling using fluorescence resonance energy transfer in living cells.
abstract: We describe and review methods for the kinetic analysis of G protein-coupled receptor (GPCR) activation and signaling that are based on optical methods. In particular, we describe the use of fluorescence resonance energy transfer (FRET) as a means of analyzing conformational changes within a single protein (for example a receptor) or between subunits of a protein complex (such as a G protein heterotrimer) and finally between distinct proteins (such as a receptor and a G protein). These methods allow the analysis of signaling kinetics in intact cells with proteins that retain their essential functional properties. They have produced a number of unexpected results: fast receptor activation kinetics in the millisecond range, similarly fast kinetics for receptor-G protein interactions, but much slower activation kinetics for G protein activation.
authors.affiliation: Institute of Pharmacology and Toxicology, University of Würzburg, D-97078 Würzburg, Germany.
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Lohse
authors.forename: Carsten
authors.initials: C
authors.lastname: Hoffmann
authors.forename: Viacheslav O
authors.initials: VO
authors.lastname: Nikolaev
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Vilardaga
authors.forename: Moritz
authors.initials: M
authors.lastname: Bünemann
mesh_terms: D000818:Animals; D002470:Cell Survival; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D020962:Heterotrimeric GTP-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D043562:Receptors, G-Protein-Coupled; D020962:Heterotrimeric GTP-Binding Proteins