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

摘要: We describe and review methods for the kinetic analysis of G protein-coupled receptor (GPCR) activation signaling that are based on optical methods. In particular, we use fluorescence resonance energy transfer (FRET) as a means analyzing conformational changes within single protein (for example receptor) or between subunits complex (such heterotrimer) finally distinct proteins protein). These allow kinetics in intact cells with retain their essential functional properties. They have produced number unexpected results: fast millisecond range, similarly receptor-G interactions, but much slower activation.