title: The involvement of protein phosphorylation in stimulus-secretion coupling in the mouse exocrine pancreas.
abstract: Secretagogue-induced protein phosphorylation was studied in the mouse pancreas in vitro, by using polyacrylamide-gel electrophoresis to separate the labelled proteins. Muscarinic cholinergic agonists increased the phosphorylation of a single band, which corresponded to Mr 32000, when the tissue was incubated with Ca2+ present in the extracellular medium, but not in Ca2+-free Krebs solution. In the presence of Ca2+, ionophore A23187 stimulated phosphorylation of the same band. The dose-response curve for carbachol-induced phosphorylation was biphasic, with maximum response at 1.0 microM-carbachol, and lesser responses when greater concentrations were used. This resembles the dose-response curve for carbachol-induced amylase secretion. The data suggest that the muscarinic-agonist-induced protein phosphorylation is stimulated secondarily to elevation of cytosol [Ca2+] and do not support the idea that diacylglycerol formed from hydrolysis of phosphatidylinositol is the activator of the protein kinase. Derivatives of cyclic AMP stimulated phosphorylation of bands corresponding to Mr 95500, 32000 and 20000. The effects of dibutyryl cyclic AMP and bethanechol on the protein of Mr 32000 were not additive, suggesting that the two agents produced phosphorylation of the same site(s) on this protein. Since derivatives of cyclic AMP, which are not very effective secretagogues in the exocrine pancreas, stimulate phosphorylation of the protein of Mr 32000, it is difficult to argue that phosphorylation of this particular protein leads to protein secretion.
mesh_terms: D000818:Animals; D018723:Bethanechol; D001626:Bethanechol Compounds / Q000494:pharmacology; D003994:Bucladesine / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002217:Carbachol / Q000494:pharmacology; D006152:Cyclic GMP / Q000031:analogs & derivatives / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D010179:Pancreas / Q000187:drug effects / Q000378:metabolism*; D010766:Phosphorylation; D011506:Proteins / Q000378:metabolism*
chemical_list: D001626:Bethanechol Compounds; D011506:Proteins; D018723:Bethanechol; C016276:8-bromocyclic GMP; D003994:Bucladesine; D002217:Carbachol; D006152:Cyclic GMP; D002118:Calcium