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. 2000 Mar;18(3):313-6.
doi: 10.1038/73767.

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo

Y Nagai  1 , M Miyazaki, R Aoki, T Zama, S Inouye, K Hirose, M Iino, M Hagiwara
Affiliations

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo

Y Nagai et al. Nat Biotechnol. 2000 Mar.

Abstract

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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