Calcium-induced inhibition of phosphoserine phosphatase in insulin target cells is mediated by the phosphorylation and activation of inhibitor 1.

Abstract

In this study, we examined the mechanism of inhibition of phosphoserine phosphatase (PSPase) activity by elevated [Ca2+]i in insulin target cells. In in vitro studies, isolated rat adipocytes were incubated with either 40 mM K+ or parathyroid hormone (PTH) (20 ng/ml) for 1 h. In in vivo studies, rats were injected with PTH (three hourly injections of 40 micrograms intraperitoneally) prior to isolation of either adipocytes or skeletal muscle. Under these conditions, intracellular [Ca2+]i changed from 100 +/- 8.7 to 263 +/- 10.5 nM. There was a concomitant 30% decrease in adipocyte PSPase activity and a 35% decrease in skeletal muscle PSPase activity, assayed using 32P-labeled phosphorylase "a" as a substrate. The inhibition of PSPase was accompanied by a 60% increase in adipocytes (p less than 0.05) and a 118% increase (p less than 0.01) in skeletal muscle inhibitor 1 (I1) activities, respectively. Since I1 is active only in the phosphorylated state, we studied the effect of [Ca2+]i on I1 phosphorylation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of heat treated extracts immunoprecipitated with I1 antibody revealed significant increase in 32P incorporation (45-60%, p less than 0.05) into I1 protein in cells with elevated [Ca2+]i. Nitrendipine, a calcium channel blocker, completely prevented increases in I1 phosphorylation and activity in cells exposed to K+ but was only partially effective in the PTH-treated cells. In contrast, a cyclic AMP antagonist, RpcAMP, prevented both the K(+)-and the PTH-induced increases in I1 phosphorylation and activity, even though it failed to block the elevations in [Ca2+]i in these cells. We conclude that [Ca2+]i-induced and cAMP-mediated phosphorylation and activation of I1 results in inhibition of PSPase activity in insulin target cells. The inhibition of PSPases may cause inappropriate serine dephosphorylation of substrates of insulin action resulting in insulin resistance.