Protein kinase C regulates the synthesis of platelet-activating factor by human monocytes.
Human peripheral blood monocytes synthesize the potent lipid autacoid platelet-activating factor (PAF) following appropriate stimulation. We examined the role of protein kinase C (PKC) in regulating the synthesis of PAF by stimulated monocytes. 4 beta-phorbol 12-myristate 13-acetate (PMA) and 1,2-dioctanoyl-sn-glycerol, which directly activate PKC, stimulated the synthesis of PAF. Sphingosine, a long-chain amine that inhibits PKC, blocked both the binding of phorbol esters to monocytes and the synthesis of PAF in response to PMA (half-maximal inhibition at 5 to 10 microM and complete inhibition at 10 to 30 microM sphingosine). Thus, the activation of PKC was necessary and sufficient for PAF synthesis in response to phorbol ester. Sphingosine also blocked PAF synthesis in response to the calcium ionophore A23187 and opsonized zymosan particles by specific inhibition of PKC. Two other PKC inhibitors, stearylamine and staurosporine, also blocked PAF synthesis following A23187 or opsonized zymosan stimulation. These experiments demonstrated that PKC activation was required for PAF synthesis in response to the calcium signal generated by A23187 or a receptor-mediated agonist, opsonized zymosan. The synthesis of PAF and leukotriene B4 were temporally coupled following cell stimulation. Further, production of these two lipid mediators, and the release of arachidonic acid, were inhibited in parallel by sphingosine. Thus, PKC regulate the synthesis of both PAF and leukotriene B4 at a common step, probably phospholipase A2.
Elstad
M R
MR
Nora Eccles Harrison Cardiovascular Research and Training Institute, University of Utah School of Medicine, Salt Lake City 84112.
McIntyre
T M
TM
Prescott
S M
SM
Zimmerman
G A
GA
D001665:Binding Sites; D000001:Calcimycin / Q000494:pharmacology; D004075:Diglycerides / Q000494:pharmacology; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D007975:Leukotriene B4 / Q000096:biosynthesis; D009000:Monocytes / Q000378:metabolism*; D009895:Opsonin Proteins; D010587:Phagocytosis; D010972:Platelet Activating Factor / Q000096:biosynthesis*; D011493:Protein Kinase C / Q000378:metabolism*; D013110:Sphingosine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000378:metabolism / Q000494:pharmacology; D015054:Zymosan / Q000494:pharmacology
D004075:Diglycerides; D009895:Opsonin Proteins; D010972:Platelet Activating Factor; C044295:1,2-dioctanoylglycerol; D007975:Leukotriene B4; D000001:Calcimycin; D015054:Zymosan; D011493:Protein Kinase C; D013110:Sphingosine; D013755:Tetradecanoylphorbol Acetate