Protein kinase A increases the binding affinity and the Ca2+ release activity of the inositol 1,4,5-trisphosphate receptor type 3 in RINm5F cells

Abstract

Background information: In endocrine cells, IP(3)R (inositol 1,4,5-trisphosphate receptor), a ligand-gated Ca2+ channel, plays an important role in the control of intracellular Ca2+ concentration. There are three subtypes of IP(3)R that are distributed differentially among cell types. RINm5F cells express almost exclusively the IP(3)R-3 subtype. The purpose of the present study was to investigate the effect of PKA (protein kinase A) on the activity of IP(3)R-3 in RINm5F cells.

Results: We show that immunoprecipitated IP(3)R-3 is a good substrate for PKA. Using a back-phosphorylation approach, we show that endogenous PKA phosphorylates IP(3)R-3 in intact RINm5F cells. [(3)H]IP(3) (inositol 1,4,5-trisphosphate) binding affinity and IP(3)-induced Ca2+ release activity were enhanced in permeabilized cells that were pre-treated with forskolin or PKA. The PKA-induced enhancement of IP(3)R-3 activity was also observed in intact RINm5F cells stimulated with carbachol and epidermal growth factor, two agonists that use different receptor types to activate phospholipase C.

Conclusion: The results of the present study reveal a converging step where the cAMP and the Ca2+ signalling systems act co-operatively in endocrine cell responses to external stimuli.