Dynamics of receptor/G protein coupling in living cells

The interaction of activated G protein‐coupled receptors with G proteins is a key event in signal transduction. Here, using a fluorescence resonance energy transfer (FRET)‐based assay, we measure directly and in living cells the interaction of YFP‐labeled α 2A ‐adrenergic receptors with CFP‐labeled...

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Veröffentlicht in:	The EMBO journal 2005-12, Vol.24 (23), p.4106-4114
Hauptverfasser:	Hein, Peter, Frank, Monika, Hoffmann, Carsten, Lohse, Martin J, Bünemann, Moritz
Format:	Artikel
Sprache:	eng
Schlagworte:	Adrenergic alpha-2 Receptor Agonists Bacterial Proteins - metabolism Cell Line Cell Membrane - metabolism EMBO37 Energy transfer Fluorescence Fluorescence Resonance Energy Transfer Fluorescent Dyes FRET G protein GIRK GTP-Binding Proteins - agonists GTP-Binding Proteins - metabolism Humans Kinetics Luminescent Proteins - metabolism Proteins Receptors, Adrenergic, alpha-2 - metabolism Receptors, G-Protein-Coupled - agonists Receptors, G-Protein-Coupled - metabolism Resonance Staining and Labeling α2A receptors
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Zusammenfassung:	The interaction of activated G protein‐coupled receptors with G proteins is a key event in signal transduction. Here, using a fluorescence resonance energy transfer (FRET)‐based assay, we measure directly and in living cells the interaction of YFP‐labeled α 2A ‐adrenergic receptors with CFP‐labeled G proteins. Upon agonist stimulation, a small, concentration‐dependent increase in FRET was observed. No specific basal FRET was detected in the absence of agonist. Kinetics of the onset of receptor/G protein interaction were
ISSN:	0261-4189 1460-2075
DOI:	10.1038/sj.emboj.7600870