Arachidonic acid-induced down-regulation of protein kinase C delta in beta-cells.
We have previously identified expression of multiple protein kinase C (PKC) isoforms in insulinoma-derived beta-cells and whole islets. Both PKC delta and PKC alpha appear to be the more abundantly expressed isoforms. In this report we studied the effects of arachidonic acid (AA) on the subcellular distribution of PKC alpha and PKC delta. AA has been reported to activate both PKC alpha and PKC delta and it is thought to be an important second messenger in beta-cells. Here we report that AA interacted with and altered beta-cell pools of PKC delta preferentially over PKC alpha. AA (100 microM) over the course of 45 min reduced cytosolic levels of PKC delta (to 40 +/- 15%, compared to time zero control) leaving membrane- and cytoskeleton-associated levels near control levels. Analysis of whole cell homogenates showed a slight down-regulation of PKC delta indicating proteolysis. The down-regulation of cytosolic PKC delta appeared to be isoform specific since cytosolic PKC alpha remained at control levels over the time course. The response was dose-dependent and negligible at concentrations below 30 microM and occurred, at least partially, in the cytosolic compartment of the cell. Indomethacin also down-regulated cytosolic PKC delta preferentially over PKC alpha possibly through accumulation of AA. These findings suggest that cytosolic PKC delta may be a downstream target of this beta-cell second messenger.
Department of Physiology and Pharmacology, College of Veterinary Medicine, University of Georgia, Athens 30602, USA.
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Knutson
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Hoenig
D000818:Animals; D016718:Arachidonic Acid / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects*; D004591:Electrophoresis, Polyacrylamide Gel; D007515:Islets of Langerhans / Q000187:drug effects / Q000201:enzymology*; D007527:Isoenzymes / Q000378:metabolism*; D008297:Male; D011493:Protein Kinase C / Q000378:metabolism*; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta; D051381:Rats
D007527:Isoenzymes; D016718:Arachidonic Acid; C497260:Prkcd protein, rat; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D051745:Protein Kinase C-delta