title: Evaluation of myosin light chain phosphorylation in isolated pancreatic acini.
abstract: The role of contractile proteins in secretory granule exocytosis was evaluated by determining whether myosin light chain phosphorylation was altered during stimulation of secretion in mouse pancreatic acini. Acinar myosin was purified by extraction into isosmotic sucrose solution containing 40 mM pyrophosphate followed by ammonium sulfate precipitation and Sepharose 4B-CL chromatography. Myosin was eluted as a single peak of K+-EDTA ATPase activity and was purified over 2,000-fold to a final ATPase specific activity of 0.96 mumol.min-1.mg protein-1. Three major myosin subunits of apparent Mr of 200,000, 20,000, and 17,000 were present in the purified myosin preparation. A fourth protein of Mr 21,000 was also present. Purification of myosin from 32P-labeled acini revealed the Mr 200,000, 21,000, and 20,000 proteins to be heavily labeled. The effect of cholecystokinin octapeptide (CCK-8) on myosin phosphorylation was studied after isolation of myosin from 32P-labeled acinar lysates by immunoprecipitation. Treatment of acini for 1-10 min with a concentration of CCK-8 that gives a maximal secretory response caused a 25-40% increase in light chain labeling. Treatment with a supramaximal CCK-8 concentration produced a 50-80% increase in light chain labeling. Phosphorylation of myosin heavy chain was not significantly affected by secretagogue treatment. These results indicate that stimulation of pancreatic acinar secretion is accompanied by an increase in myosin light chain phosphorylation.
authors.affiliation: Department of Physiology, University of California, San Francisco 94143.
authors.forename: D B
authors.initials: DB
authors.lastname: Burnham
authors.forename: H D
authors.initials: HD
authors.lastname: Söling
authors.forename: J A
authors.initials: JA
authors.lastname: Williams
mesh_terms: D000818:Animals; D066298:In Vitro Techniques; D051379:Mice; D008815:Mice, Inbred Strains; D009218:Myosins / Q000378:metabolism*; D010179:Pancreas / Q000378:metabolism*; D010766:Phosphorylation; D012844:Sincalide / Q000494:pharmacology
chemical_list: D009218:Myosins; D012844:Sincalide