siRNA-mediated depletion of endogenous protein phosphatase 2Acα markedly attenuates ceramide-activated protein phosphatase activity in insulin-secreting INS-832/13 cells

The sphingolipid ceramide (CER) and its metabolites have been recognized as important mediators of signal transduction processes leading to a variety of cellular responses, including survival and demise via apoptosis. Accumulating evidence implicates key regulatory roles for intracellularly generated CER in metabolic dysfunction of the islet β cell. We have previously reported localization of an okadaic (OKA)-sensitive CER-activated protein phosphatase (CAPP) in the islet β cell. We have also reported immunological identification of the structural A subunit, the regulatory B56α subunit, and the catalytic C subunit for CAPP holoenzyme complex in insulin-secreting INS-1 cells. Herein, we provide the first evidence to suggest that siRNA-mediated knockdown of the α isoform of the catalytic subunit of PP2Ac (PP2Acα) markedly reduces the CAPP activity in INS 832/13 cells. Potential significance of the functional activation of CAPP holoenzyme in the context of lipid-and glucose-induced metabolic dysfunction of the islet β cell is discussed.