Phorbol ester-induced inhibition of the beta-adrenergic system in pulmonary endothelium: role of a pertussis toxin-sensitive protein.
To investigate possible cellular mechanisms for how activation of protein kinase C inhibits the relaxation caused by isoproterenol, we studied the effect of the protein kinase C activator 4 beta-phorbol-12 beta-myristate-13 alpha-acetate (PMA) on the increase in cyclic AMP (cAMP) production and adenylate cyclase activity caused by isoproterenol in bovine pulmonary artery endothelial cells. Treatment of intact cells with PMA prevented in a time- and dose-dependent manner the increase in cAMP production caused by isoproterenol, whereas 4 alpha-phorbol-12 beta-myristate-13 alpha-acetate (4 alpha-PMA), which does not activate protein kinase C, did not affect isoproterenol-induced cAMP production. PMA also reduced the increase in adenylate cyclase activity caused by isoproterenol, forskolin, and Gpp(NH)p. To test the hypothesis that the inhibitory effect of PMA is mediated via a pertussis toxin-sensitive G protein, we determined whether pretreatment of the cells with pertussis toxin would prevent the inhibitory effects of PMA. In pulmonary endothelial cells, pertussis toxin ADP-ribosylated an Mr 40,000 peptide that comigrated with the pertussis toxin substrate of human erythrocytes. Pertussis toxin treatment eliminated the inhibitory effect of PMA on isoproterenol-stimulated cAMP production and adenylate cyclase activity. Thus, the protein kinase C activator PMA inhibits the increase in cAMP production and adenylate cyclase caused by isoproterenol. This inhibitory effect in endothelial cells appears to be mediated via a pertussis toxin-sensitive protein.
Newman
K B
KB
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland.
Michael
J R
JR
Feldman
A M
AM
D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000096:biosynthesis; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism*; D007545:Isoproterenol / Q000494:pharmacology; D037342:Pertussis Toxin; D011493:Protein Kinase C / Q000378:metabolism; D011651:Pulmonary Artery / Q000166:cytology / Q000378:metabolism*; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology*
D037361:Adenylate Cyclase Toxin; D011943:Receptors, Adrenergic, beta; D010566:Virulence Factors, Bordetella; D000242:Cyclic AMP; D037342:Pertussis Toxin; D011493:Protein Kinase C; D019204:GTP-Binding Proteins; D000262:Adenylyl Cyclases; D007545:Isoproterenol; D013755:Tetradecanoylphorbol Acetate