title: Catalytic subunit of protein kinase A induces amylase release from streptolysin O-permeabilized parotid acini.
abstract: Amylase release from parotid acinar cells is a typical model of cAMP-mediated exocytosis. To obtain unequivocal data concerning the role of cAMP-dependent protein kinase (PKA) in amylase exocytosis, we undertook the direct introduction of the PKA catalytic subunit into the parotid acini by permeabilization with streptolysin O (SLO). In the presence of 100 hemolytic units/ml SLO, cAMP increased amylase release in a time- and dose-dependent manner. PKI-(5-24)-peptide, a specific PKA inhibitor, markedly inhibited amylase release, but the extent of inhibition was approximately 50%. On the other hand, the PKA catalytic subunit highly purified from bovine hearts significantly induced amylase release. The release was strictly dependent on the presence of SLO and the catalytic activity of PKA added. The catalytic subunit dose dependently induced amylase release, but the heat-inactivated subunit had no stimulatory effect. PKI-(5-24)-peptide completely blocked amylase release evoked by the subunit. These results clearly demonstrate that the catalytic subunit of PKA regulates cAMP-mediated amylase release through phosphorylation of unidentified protein(s) directly or indirectly involved in the process of exocytosis.
authors.affiliation: Department of Oral Biochemistry, School of Dentistry, Health Sciences University of Hokkaido, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Takuma
authors.forename: T
authors.initials: T
authors.lastname: Ichida
mesh_terms: D000681:Amylases / Q000096:biosynthesis / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins; D002384:Catalysis; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004790:Enzyme Induction; D005089:Exocytosis; D010306:Parotid Gland / Q000187:drug effects / Q000201:enzymology*; D051381:Rats; D013301:Streptolysins / Q000494:pharmacology
chemical_list: D001426:Bacterial Proteins; D013301:Streptolysins; C025964:streptolysin O; D000242:Cyclic AMP; D017868:Cyclic AMP-Dependent Protein Kinases; D000681:Amylases