Lipopolysaccharide in combination with serum potentiates the stimulated activity of phospholipase D in human neutrophils via CD14.
The addition of fMet-Leu-Phe or phorbol 12-myristate 13-acetate to human neutrophils stimulates phospholipase D activity as evidenced by the release of phosphatidic acid and the generation of diacylglycerol, and in the presence of ethanol the formation of phosphatidyl ethanol. The activation of phospholipase D by either the chemotactic factor or active phorbol ester is inhibited by the tyrosine kinase inhibitor erbstatin. The fMet-Leu-Phe-induced stimulation of this enzyme is greatly potentiated in cells which have been preincubated with low concentrations of lipopolysaccharide and serum. The presence of serum is essential for the potentiation by low concentrations of lipopolysaccharide. Moreover, the monoclonal antibody MY4(IgG2b) against CD14 inhibits the potentiation by the low concentration of lipopolysaccharide. These data suggest three important points. First, a tyrosine kinase step is necessary for the activation of phospholipase D. This suggests that the phospholipase D enzyme needs to be phosphorylated on tyrosine residues to be activated. Second, low concentrations of lipopolysaccharide, in the presence of serum, can potentiate the stimulated activity of this enzyme. Third, the priming action of the lipopolysaccharide-serum complex is mediated by CD14.
Yasui
K
K
Department of Physiology, University of Connecticut Health Center, Farmington 06030.
Becker
E L
EL
Sha'afi
R I
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D000328:Adult; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D015703:Antigens, CD / Q000502:physiology*; D015214:Antigens, Differentiation, Myelomonocytic / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D004075:Diglycerides / Q000096:biosynthesis; D004357:Drug Synergism; D004789:Enzyme Activation; D000431:Ethanol / Q000378:metabolism / Q000494:pharmacology; D020404:Glycerophospholipids; D006801:Humans; D006873:Hydroquinones / Q000494:pharmacology; D007424:Intracellular Fluid / Q000378:metabolism; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008969:Molecular Sequence Data; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology* / Q000276:immunology; D010712:Phosphatidic Acids / Q000096:biosynthesis / Q000378:metabolism; D010739:Phospholipase D / Q000187:drug effects / Q000378:metabolism*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D016897:Respiratory Burst / Q000502:physiology; D013268:Stimulation, Chemical; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D004075:Diglycerides; D020404:Glycerophospholipids; D006873:Hydroquinones; D018950:Lipopolysaccharide Receptors; D008070:Lipopolysaccharides; D010712:Phosphatidic Acids; C051521:phosphatidylethanol; D000431:Ethanol; D009240:N-Formylmethionine Leucyl-Phenylalanine; D011505:Protein-Tyrosine Kinases; D010739:Phospholipase D; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium; C048212:erbstatin