Format

Send to

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

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

Author information

1
Department of Functional Genomics, Medical Research Institute, Tokyo Medical and Dental University, Yushima, Tokyo 113-8510, Japan.

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.

PMID:
10700148
DOI:
10.1038/73767
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Nature Publishing Group
Loading ...
Support Center