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Nat Biotechnol. 2000 Mar;18(3):313-6.

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo.

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Department of Functional Genomics, Medical Research Institute, Tokyo Medical and Dental University, Yushima, Tokyo 113-8510, Japan.


We have developed a method for visualizing phosphorylation of proteins in living cells using a novel fluorescent indicator composed of two green fluorescent protein (GFP) variants joined by the kinase-inducible domain (KID) of the transcription factor cyclic adenosine monophosphate (cAMP)-responsive element binding protein (CREB). Phosphorylation of KID by the cAMP-dependent protein kinase A (PKA) decreased the fluorescence resonance energy transfer (FRET) among the flanking GFPs. By transfecting COS-7 cells with an expression vector encoding this indicator protein (termed ART for cAMP-responsive tracer), we were able to visualize activation dynamics of PKA in living cells.

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