Long-chain acyl CoA regulation of protein kinase C and fatty acid potentiation of glucose-stimulated insulin secretion in clonal beta-cells.
Pancreatic beta-cells contain protein kinase C (PKC) isoforms that may play a role in insulin secretion. Activity of PKC classes (cPKC, nPKC, aPKC) and their regulation by acyl-CoA derivatives was examined in extracts of clonal pancreatic beta-cells (HIT) by protein phosphorylation. PKC classes were distinguished based on their previously defined cofactor requirements. Down-regulation of PKC by phorbol esters was confirmed by Western blotting and resulted in the complete loss of cPKC activity, partial loss of nPKC activity and preservation of aPKC activity and glucose-stimulated insulin secretion. aPKC activity was potentiated 4- to 8-fold by the CoA esters of myristate, palmitate, and oleate with a half-maximal value of 3 microM. Both oleoyl- and myristol-CoA, but not palmitoyl-CoA, caused inhibition of nPKC activity. Oleoyl-CoA inhibited nPKC activity up to 75% with a half-maximal effect at 10 microM. This value was independent of the concentration of diacylglycerol used. The addition of exogenous oleate or palmitate potentiated glucose-stimulated insulin secretion 2-fold and was unaffected by PMA-induced down-regulation. Stimulation by glucose or glucose and oleate also increased the mass of PKC-zeta found in the particulate fraction. These data are consistent with increased cytosolic long-chain acylCoA-activating aPKC isoforms resulting in stimulation and/or potentiation of glucose-induced insulin secretion.
Yaney
G C
GC
Obesity Research Center, Evans Department of Medicine, Boston University School of Medicine, Massachusetts 02118, USA. gyaney@acs.bu.edu
Korchak
H M
HM
Corkey
B E
BE
D000214:Acyl Coenzyme A / Q000494:pharmacology*; D015153:Blotting, Western; D002460:Cell Line; D002999:Clone Cells; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005227:Fatty Acids / Q000494:pharmacology*; D005947:Glucose / Q000494:pharmacology*; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000187:drug effects / Q000378:metabolism*; D019814:Myristic Acid / Q000494:pharmacology; D019301:Oleic Acid / Q000494:pharmacology; D019308:Palmitic Acid / Q000494:pharmacology; D010171:Palmitoyl Coenzyme A / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*
D000214:Acyl Coenzyme A; D004791:Enzyme Inhibitors; D005227:Fatty Acids; D007328:Insulin; D019814:Myristic Acid; C017585:oleoyl-coenzyme A; D010171:Palmitoyl Coenzyme A; D019301:Oleic Acid; D019308:Palmitic Acid; C040349:S-tetradecanoyl-coenzyme A; D011493:Protein Kinase C; D005947:Glucose