“Shaping” of cell signaling via AKAP-tethered PDE4D: Probing with AKAR2-AKAP5 biosensor
PKA, a key regulator of cell signaling, phosphorylates a diverse and important array of target molecules and is spatially docked to members of the A-kinase Anchoring Protein (AKAP) family. AKAR2 is a biosensor which yields a FRET signal in vivo, when phosphorylated by PKA. AKAP5, a prominent member of the AKAP family, docks several signaling molecules including PKA, PDE4D, as well as GPCRs, and is obligate for the propagation of the activation of the mitogen-activated protein kinase cascade from GPCRs to ERK1,2.
Using an AKAR2-AKAP5 fusion “biosensor”, we investigated the spatial-temporal activation of AKAP5 undergoing phosphorylation by PKA in response to β-adrenergic stimulation. The pattern of PKA activation reported by AKAR2-AKAP5 is a more rapid and spatially distinct from those “sensed” by AKAR2-AKAP12. Spatial-temporal restriction of activated PKA by AKAP5 was found to “shape” the signaling response. Phosphatase PDE4D tethered to AKAP5 also later reverses within 60 s elevated intracellular cyclic AMP levels stimulated by β-adrenergic agonist. AKAP12, however, fails to attenuate the rise in cyclic AMP over this time. Fusion of the AKAP5 PDE4D-binding-domain to AKAP12 was found to accelerate a reversal of accumulation of intracellular cyclic AMP.
AKAPs, which are scaffolds with tethered enzymes, can “shape” the temporal and spatial aspects of cell signaling.
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- “Shaping” of cell signaling via AKAP-tethered PDE4D: Probing with AKAR2-AKAP5 biosensor
- Open Access
- Available under Open Access This content is freely available online to anyone, anywhere at any time.
Journal of Molecular Signaling
- Online Date
- May 2012
- Online ISSN
- BioMed Central
- Additional Links
- β-adrenergic receptor
- Protein kinase A
- Author Affiliations
- 1. Department of Pharmacological Sciences, Health Sciences Center, BST-7, SUNY at Stony Brook, School of Medicine, Stony Brook, New York, 11794-8651, USA
- 2. Department of Physiology and Biophysics, SUNY at Stony Brook, School of Medicine, Stony Brook, New York, 11794-8661, USA