"cAMP sponge": a buffer for cyclic adenosine 3', 5'-monophosphate.

ABSTRACT:

Background

While intracellular buffers are widely used to study calcium signaling, no such tool exists for the other major second messenger, cyclic AMP (cAMP).

Methods/principal findings

Here we describe a genetically encoded buffer for cAMP based on the high-affinity cAMP-binding carboxy-terminus of the regulatory subunit RIbeta of protein kinase A (PKA). Addition of targeting sequences permitted localization of this fragment to the extra-nuclear compartment, while tagging with mCherry allowed quantification of its expression at the single cell level. This construct (named "cAMP sponge") was shown to selectively bind cAMP in vitro. Its expression significantly suppressed agonist-induced cAMP signals and the downstream activation of PKA within the cytosol as measured by FRET

SUBMITTER: Lefkimmiatis K 

PROVIDER: S-EPMC2766031 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature