Interactions of okadaic acid with insulin action in hepatocytes: role of protein phosphatases in insulin action.
The regulation of carbohydrate metabolism involves changes in the phosphorylation state of enzymes. We used okadaic acid, a potent inhibitor of protein phosphatases type 2A (IC50 0.05-2 nM) and type 1 (IC50 10-20 nM) to determine the role of these phosphatases in the control of carbohydrate metabolism by insulin in rat hepatocytes. In the absence of insulin, okadaic acid caused total inhibition of glycogen synthesis at 100 nM and half-maximal inhibition at 8-9 nM. In the presence of insulin, lower concentrations of okadaic acid (to which type 2A phosphatases are sensitive) were effective at inhibiting glycogen synthesis. 2.5 nM okadaic acid caused total inhibition of the 2-fold stimulation of glycogen synthesis by insulin but had no effect on the basal unstimulated rate of glycogen synthesis. This suggests the involvement of type 2A protein phosphatases in the stimulation of glycogen synthesis by insulin. Okadaic acid (5 nM), partially suppressed but did not abolish the increase in glucokinase mRNA levels caused by insulin, indicating that dephosphorylation mechanisms may be involved in the control of glucokinase mRNA levels by insulin. It is concluded that activation of protein phosphatases type 1 and/or type 2A by insulin may have a widespread role in the control of glucose metabolism at various sites.
Agius
L
L
Department of Medicine, Medical School, University of Newcastle-upon-Tyne, U.K.
Peak
M
M
D000818:Animals; D002478:Cells, Cultured; D004988:Ethers, Cyclic / Q000494:pharmacology*; D015870:Gene Expression / Q000187:drug effects; D005941:Glucokinase / Q000235:genetics; D066298:In Vitro Techniques; D007328:Insulin / Q000494:pharmacology*; D007329:Insulin Antagonists / Q000494:pharmacology; D008099:Liver / Q000378:metabolism*; D008112:Liver Glycogen / Q000096:biosynthesis*; D008297:Male; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011919:Rats, Inbred Strains
D004988:Ethers, Cyclic; D007328:Insulin; D007329:Insulin Antagonists; D008112:Liver Glycogen; D012333:RNA, Messenger; D019319:Okadaic Acid; D005941:Glucokinase; D010749:Phosphoprotein Phosphatases