Certain inhibitors of protein serine/threonine kinases also inhibit tyrosine phosphorylation of phospholipase C gamma 1 and other proteins and reveal distinct roles for tyrosine kinase(s) and protein kinase C in stimulated, rat basophilic RBL-2H3 cells.
Various inhibitors of phospholipases and serine/threonine kinases were used to determine whether activation of these enzymes was necessary for Ag-induced exocytosis in rat basophilic RBL-2H3 cells. Several inhibitors, however, inhibited events other than those intended in stimulated RBL-2H3 cells. Staurosporine and KT5926, inhibitors of protein kinase C and myosin L chain kinase, respectively, suppressed, in a dose-dependent manner, hydrolysis of inositol phospholipids, release of arachidonic acid, and exocytosis in cells stimulated with Ag or Ca(2+)-ionophore, A23187. Such generalized inhibition could also be induced in permeabilized cells with several peptide inhibitors of tyrosine kinases. All the above inhibitors suppressed Ag-induced tyrosine phosphorylation of several proteins, including phospholipase C gamma 1, and this suppression correlated with the inhibition of hydrolysis of inositol phospholipids and exocytosis. Three inhibitors of protein kinase C, Ro31-7549, calphostin C, and a peptide inhibitor, did not inhibit the tyrosine phosphorylation of proteins but selectively blocked exocytosis, presumably, by inhibiting protein kinase C. Thus, both tyrosine phosphorylation of proteins and the activation of protein kinase C were necessary events for hydrolysis of inositol phospholipids and exocytosis.
Yamada
K
K
Laboratory of Chemical Pharmacology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892.
Jelsema
C L
CL
Beaven
M A
MA
D000470:Alkaloids / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D000803:Angiotensin I / Q000494:pharmacology; D000818:Animals; D000941:Antigens; D016718:Arachidonic Acid / Q000378:metabolism; D001491:Basophils / Q000201:enzymology*; D000001:Calcimycin / Q000494:pharmacology; D002227:Carbazoles; D005089:Exocytosis / Q000187:drug effects; D066298:In Vitro Techniques; D007211:Indoles / Q000494:pharmacology; D007295:Inositol Phosphates / Q000378:metabolism; D007942:Leukemia, Experimental; D008301:Maleimides / Q000494:pharmacology; D010766:Phosphorylation; D019000:Phosphotyrosine; D011493:Protein Kinase C / Q000378:metabolism*; D047428:Protein Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors*; D051381:Rats; D012701:Serotonin / Q000378:metabolism; D019311:Staurosporine; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000378:metabolism*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D000077191:Wortmannin
D000470:Alkaloids; D000730:Androstadienes; D000941:Antigens; D002227:Carbazoles; D007211:Indoles; D007295:Inositol Phosphates; D008301:Maleimides; D047428:Protein Kinase Inhibitors; C063769:KT 5926; D019000:Phosphotyrosine; D016718:Arachidonic Acid; D012701:Serotonin; D000001:Calcimycin; D014443:Tyrosine; D000803:Angiotensin I; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases; D011493:Protein Kinase C; D010738:Type C Phospholipases; D019311:Staurosporine; D000077191:Wortmannin; C064755:Ro 31-7549