Research article

Journal of Molecular Signaling

, 7:4

First online:

Open Access This content is freely available online to anyone, anywhere at any time.

“Shaping” of cell signaling via AKAP-tethered PDE4D: Probing with AKAR2-AKAP5 biosensor

  • Salih S KoçerAffiliated withDepartment of Pharmacological Sciences, Health Sciences Center, BST-7, SUNY at Stony Brook, School of Medicine, Stony Brook Email author 
  • , Hsien-yu WangAffiliated withDepartment of Physiology and Biophysics, SUNY at Stony Brook, School of Medicine, Stony Brook
  • , Craig C MalbonAffiliated withDepartment of Pharmacological Sciences, Health Sciences Center, BST-7, SUNY at Stony Brook, School of Medicine, Stony Brook

Abstract

Background

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.

Results

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.

Conclusion

AKAPs, which are scaffolds with tethered enzymes, can “shape” the temporal and spatial aspects of cell signaling.

Keywords

AKAP5 AKAP12 AKAR2 β-adrenergic receptor PDE4D Protein kinase A Scaffold Tethered