TRANSLOCATION OF THE ALPHA-ISOZYME OF PROTEIN-KINASE-C DURING STIMULATION OF RAT PAROTID ACINAR-CELLS BY PHORBOL ESTER AND CARBACHOL

Protein kinase C activity was detected in the cytosolic fraction of quiescent parotid acinar cells; the particulate fraction contained a much smaller proportion of the enzyme. Protein kinase C activity was increased in the membrane fraction and decreased in the cytosol after exposure of intact cells to phorbol 12-myristate 13-acetate (PMA) or the muscarinic-receptor agonist carbachol. The effect of PMA was potentiated by a subthreshold concentration of ionomycin. Immunoblot analysis with anti-protein kinase C antibodies revealed that the protein kinase C-alpha isoform is expressed in rat parotid cells. Other Ca2+-dependent isoforms were not detected. Further, agonist stimulation caused the redistribution of protein kinase C-alpha from cytosol to a membrane fraction. Agonists may promote parotid acinar cell activity, including amylase secretion, by increasing the affinity of protein kinase C-alpha for the membrane fraction, presumably via a rise in Ca2+ and diacylglycerol derived from polyphosphoinositide hydrolysis.