doi: 10.1038/73767.
A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo
Affiliations
- PMID: 10700148
- DOI: 10.1038/73767
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A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo
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.
Comment in
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Novel fluorescent approaches for studying cell signaling in single cells.Nat Biotechnol. 2000 Mar;18(3):262-3. doi: 10.1038/73687. Nat Biotechnol. 2000. PMID: 10700133 No abstract available.
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