Glucose metabolites inhibit protein phosphatases and directly promote insulin exocytosis in pancreatic beta-cells.
In human type 2 diabetes mellitus, loss of glucose-sensitive insulin secretion is an early pathogenetic event. Glucose is the cardinal physiological stimulator of insulin secretion from the pancreatic beta-cell, but the mechanisms involved in glucose sensing are not fully understood. Specific ser/thr protein phosphatase (PPase) inactivation by okadaic acid promotes Ca(2+) entry and insulin exocytosis in the beta-cell. We now show that glycolytic and Krebs cycle intermediates, whose concentrations increase upon glucose stimulation, not only dose dependently inhibit ser/thr PPase enzymatic activities, but also directly promote insulin exocytosis from permeabilized beta-cells. Thus, fructose-1,6-bisphosphate, phosphoenolpyruvate, 3-phosphoglycerate, citrate, and oxaloacetate inhibit PPases and significantly enhance insulin exocytosis, nonadditive to that of okadaic acid, at micromolar Ca2+ concentrations. In contrast, the effect of GTP is potentiated by okadaic acid, suggesting that the action of GTP does not require PPase inactivation. We conclude that specific glucose metabolites and GTP inhibit beta-cell PPase activities and directly stimulate Ca2+-independent insulin exocytosis. The glucose metabolites, but not GTP, seem to require PPase inactivation for their stimulatory effect on exocytosis. Thus, an increase in phosphorylation state, through inhibition of protein dephosphorylation by metabolic intermediates, may be a novel regulatory mechanism linking glucose sensing to insulin exocytosis in the beta-cell.
Sjöholm
Ake
A
Department of Internal Medicine, Karolinska Institutet, Stockholm South Hospital, SE 118 83 Stockholm, Sweden.
Lehtihet
Mikael
M
Efanov
Alexandre M
AM
Zaitsev
Sergei V
SV
Berggren
Per-Olof
PO
Honkanen
Richard E
RE
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D019343:Citric Acid / Q000494:pharmacology; D004357:Drug Synergism; D004791:Enzyme Inhibitors / Q000378:metabolism* / Q000494:pharmacology*; D005089:Exocytosis / Q000187:drug effects*; D005635:Fructosediphosphates / Q000494:pharmacology; D005947:Glucose / Q000378:metabolism*; D005988:Glyceric Acids / Q000494:pharmacology; D006160:Guanosine Triphosphate / Q000494:pharmacology; D007328:Insulin / Q000378:metabolism*; D007340:Insulinoma; D007515:Islets of Langerhans / Q000378:metabolism*; D019319:Okadaic Acid / Q000494:pharmacology; D062907:Oxaloacetic Acid / Q000494:pharmacology; D010190:Pancreatic Neoplasms; D010728:Phosphoenolpyruvate / Q000494:pharmacology; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010766:Phosphorylation; D051381:Rats; D014407:Tumor Cells, Cultured
D004791:Enzyme Inhibitors; D005635:Fructosediphosphates; D005988:Glyceric Acids; D007328:Insulin; D019319:Okadaic Acid; D019343:Citric Acid; D062907:Oxaloacetic Acid; D010728:Phosphoenolpyruvate; C005156:3-phosphoglycerate; D006160:Guanosine Triphosphate; D010749:Phosphoprotein Phosphatases; D005947:Glucose; C029063:fructose-1,6-diphosphate; D002118:Calcium