Role of Ca2+-independent phospholipase A2 in exocytosis of amylase from parotid acinar cells.
We evaluated the role of cytosolic phospholipase A2 (PLA2) in the exocytosis of amylase from parotid acinar cells. The exocytosis stimulated by isoproterenol was dose-dependently inhibited by bromoenol lactone (BEL), a potent suicide inhibitor of Ca2+-independent cytosolic PLA2. The IC50 value of BEL was approximately 7 microM. AACOCF3, a selective inhibitor of Ca2+-dependent cytosolic PLA2, did not inhibit the exocytosis at least up to 30 microM. BEL also inhibited amylase release evoked by forskolin and membrane-permeable cAMP, but it did not inhibit cAMP-dependent protein kinase activity. PLA2 activity in parotid acinar cells was found to be predominantly Ca2+-independent, and was strongly inhibited by BEL, whose IC50 value was approximately 2 microM when it was applied to intact acini. Although isoproterenol scarcely enhanced [3H]arachidonic acid release from intact acinar cells, BEL dose-dependently decreased the basal arachidonic acid release to approximately one half of the control value. These results suggest that the cytosolic Ca2+-independent PLA2 activity plays a role in the membrane fusion process of exocytosis in parotid acinar cells.
Takuma
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T
Department of Oral Biochemistry, School of Dentistry, Health Sciences University of Hokkaido, Tobetsu. takuma@hucc.hokudai.ac.jp
Ichida
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T
D000681:Amylases / Q000378:metabolism*; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D003600:Cytosol / Q000201:enzymology; D005089:Exocytosis / Q000502:physiology*; D054522:Group VI Phospholipases A2; D007545:Isoproterenol / Q000494:pharmacology; D009281:Naphthalenes / Q000494:pharmacology; D010306:Parotid Gland / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D010741:Phospholipases A / Q000502:physiology*; D054467:Phospholipases A2; D011753:Pyrones / Q000494:pharmacology; D051381:Rats
D009281:Naphthalenes; D010726:Phosphodiesterase Inhibitors; D011753:Pyrones; D016718:Arachidonic Acid; C068314:6-(bromomethylene)tetrahydro-3-(1-naphthaleneyl)-2H-pyran-2-one; D010741:Phospholipases A; D054522:Group VI Phospholipases A2; D054467:Phospholipases A2; D000681:Amylases; D007545:Isoproterenol