Protein phosphorylation in pancreatic islets induced by 3-phosphoglycerate and 2-phosphoglycerate.
We have shown previously that 3-phosphoglycerate, which is a glycolytic metabolite of glucose, induces protein phosphorylation in bovine and rat brain and in rat heart, kidney, liver, lung, and whole pancreas. Since glycolytic metabolism of glucose is of paramount importance in insulin release, we considered the possibility that 3-phosphoglycerate may act as a coupling factor, and we searched for evidence for the existence of 3-phosphoglycerate-dependent protein phosphorylation systems in freshly isolated normal rat pancreatic islets. Membrane and cytosol fractions were incubated with [gamma-32P]ATP and appropriate test substances and were subjected to NaDodSO4/PAGE and autoradiography. As little as 0.005 mM 3-phosphoglycerate or 2-phosphoglycerate stimulated the phosphorylation of a 65-kDa cytosol protein by as early as 0.25 min. The phosphate bond of the 65-kDa phosphoprotein was sufficiently stable to withstand dialysis; the radioactivity could not be chased out by subsequent exposure to ATP, ADP, 3-phosphoglycerate, or 2,3-bisphosphoglycerate. Moreover, cAMP, cGMP, phorbol 12-myristate 13-acetate, or calcium failed to stimulate the phosphorylation of the 65-kDa protein. Phosphoglycerate-dependent protein phosphorylation in islets may have relevance to stimulation of insulin secretion.
Pek
S B
SB
Department of Internal Medicine, University of Michigan, Ann Arbor 48109.
Usami
M
M
Bilir
N
N
Fischer-Bovenkerk
C
C
Ueda
T
T
D001731:Bisphosphoglycerate Mutase / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D006152:Cyclic GMP / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005988:Glyceric Acids / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism; D008970:Molecular Weight; D010750:Phosphoproteins / Q000378:metabolism*; D011493:Protein Kinase C / Q000502:physiology; D011494:Protein Kinases / Q000378:metabolism*; D013997:Time Factors
D005988:Glyceric Acids; D008565:Membrane Proteins; D010750:Phosphoproteins; C008885:2-phosphoglycerate; C005156:3-phosphoglycerate; D000242:Cyclic AMP; D011494:Protein Kinases; D011493:Protein Kinase C; D001731:Bisphosphoglycerate Mutase; D006152:Cyclic GMP; D002118:Calcium