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Endogenous substrate proteins for calcium calmodulin dependent calcium phospho lipid dependent and cyclic amp dependent protein kinases in mouse pancreatic islets



Endogenous substrate proteins for calcium calmodulin dependent calcium phospho lipid dependent and cyclic amp dependent protein kinases in mouse pancreatic islets



Biochemical Journal 221(1): 247-254



The occurrence of endogenous substrate proteins for Ca2+-dependent protein kinase, augmented by either phospholipid or calmodulin, and for cAMP-dependent protein kinase was examined in homogenates and subcellular fraction of mouse pancreatic islets. Islet protein phosphorylation was enhanced by Ca2+-calmodulin; the major endogenous substrates in the homogenate were 2 proteins of MW 53,000 and 100,000. The MW-100,000 phosphoprotein was localized to a 27,000g-supernatant fraction, whereas the MW-53,000 phosphoprotein was present in a 27,000g particulate fraction of mouse islets. In the presence of Ca2+, phosphatidylserine stimulated phosphorylation of 15 proteins, of MW 17,000-190,000, in a 27,000g-supernatant fraction. No effects of Ca2+ plus phosphatidylserine were observed in a 27,000g particulate fraction of mouse islets. Examination of cAMP-dependent protein phosphorylation revealed 5 substrate proteins, of MW 23,000-72,000, present in the 27,000g supernatant of mouse islets. No common substrates for either the 2 Ca2+-dependent phosphorylation systems or for the cAMP-dependent and the Ca2+-calmodulin-dependent phosphorylation systems were noted. The actions of the cAMP-sensitive and the Ca2+-phospholipid-sensitive systems may be overlapping, since 2 common substrates for them were noted in the 27,000g-supernatant fraction. The results are consistent with the hypothesis that protein phosphorylation may play a role in the regulation of insulin secretion by Ca2+ and cAMP. The extensive stimulatory effect of phosphatidylserine furthermore suggests that the Ca2+-phospholipid-sensitive protein kinase may prove to be a prominent phosphorylation system in pancreatic islets.

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Endogenous substrate proteins for Ca2+-calmodulin-dependent, Ca2+-phospholipid-dependent and cyclic AMP-dependent protein kinases in mouse pancreatic islets. Biochemical Journal 221(1): 247-253, 1984

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