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Adv Protein Chem. 2007;74:167-88.

Kinetic analysis of G protein-coupled receptor signaling using fluorescence resonance energy transfer in living cells.

Author information

1
Institute of Pharmacology and Toxicology, University of Würzburg, D-97078 Würzburg, Germany.

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.

PMID:
17854658
DOI:
10.1016/S0065-3233(07)74005-6
[Indexed for MEDLINE]

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