Enhancement of cyclic AMP metabolism in a B cell line by protein kinase C.
In a human B cell line in which we previously demonstrated an inverse relationship between cyclic adenosine monophosphate (cAMP) content and immunoglobulin secretion, the phorbol ester, phorbol myristate acetate, (PMA), was shown to augment the cAMP elevating ability of cholera toxin (CT), suggesting a regulatory linkage between the two transmembrane signaling pathways, cAMP and phospholipid (J. Immunol. 141, 1678-1686, 1988). We now extend these studies and provide additional evidence that activated protein kinase C, a principal product of the activation of the hydrolytic phospholipid pathway, plays a direct role in the augmentation of cAMP levels in cells stimulated by diverse cAMP-elevating ligands. Prostaglandin E1 (PGE1), forskolin (FSK) and CT, all of which demonstrated a concentration and time-dependent elevation of intracellular cAMP, produced even greater (up to twofold) elevations of cAMP in the presence of PMA or the diacylglycerol analogs, 1,2-dioctanoylglycerol (DiC8), and 1-oleoyl-2-acetylglycerol (OAG). In the absence of CT, PGE1, or FSK, these protein kinase C activators produced only small increases in cAMP content of the cells. Several tests of protein kinase C specificity in these PMA-, DiC8-, and OAG-induced augmentations were made: (i) only phorbol esters known to activate protein kinase C worked, (ii) PMA augmentation was abolished by down-regulation of protein kinase C, (iii) Staurosporine (a known inhibitor of protein kinase C) selectively inhibited the effects of PMA on cAMP generation and on immunoglobulin secretion in the LA350 cell line.
Patke
C L
CL
Department of Pediatrics, Baylor College of Medicine, Houston, Texas 77030.
Shearer
W T
WT
D000470:Alkaloids / Q000494:pharmacology; D000527:Alprostadil / Q000494:pharmacology; D001402:B-Lymphocytes / Q000502:physiology*; D002460:Cell Line; D002772:Cholera Toxin / Q000494:pharmacology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism*; D004075:Diglycerides / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects; D004357:Drug Synergism; D006801:Humans; D007075:Immunoglobulin M / Q000378:metabolism; D066298:In Vitro Techniques; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D015398:Signal Transduction; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000470:Alkaloids; D004075:Diglycerides; D007075:Immunoglobulin M; D005576:Colforsin; D002772:Cholera Toxin; D000242:Cyclic AMP; D011493:Protein Kinase C; D000527:Alprostadil; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate