Tyrosine phosphorylation of 100-115 kDa proteins by phosphatidic acid generated via phospholipase D activation in HL60 granulocytes.
In HL60 granulocytes, 4beta-phorbol 12-myristate 13-acetate (PMA) induced tyrosine phosphorylation of several proteins with molecular weight of 100-115 kDa and 45 kDa. Furthermore, PMA-mediated phosphatidic acid (PA) production via phospholipase D (PLD) activation. In the presence of either butanol or ethanol, PMA-induced PA production was markedly reduced and instead a metabolically stable phosphatidylbutanol (PBut) or phosphatidylethanol (PEt) was produced by transphosphatidylation by PLD. Under the same incubation condition, these primary alcohols inhibited PMA-induced tyrosine phosphorylation of the 100-115 kDa proteins. Propranolol, which is often used as a selective inhibitor of PA phosphohydrolase (PAP) involving diacylglycerol (DG) formation from PA, did not affect tyrosine phosphorylation of the 100-115 kDa proteins. Moreover, incubation of HL60 granulocytes with Streptomyces chromofuscus PLD caused both PA production and tyrosine phosphorylation of the above proteins. Exogenous PA treatment also induced tyrosine phosphorylation of the same proteins. Thus, the results presented here suggest that PA produced via PLD activation is involved in tyrosine phosphorylation of the 100-115 kDa proteins in HL60 granulocytes.
Ohguchi
K
K
Department of Biochemistry, Gifu University School of Medicine, Japan.
Kasai
T
T
Nozawa
Y
Y
D015153:Blotting, Western; D000440:Butanols / Q000378:metabolism / Q000494:pharmacology; D004789:Enzyme Activation; D000431:Ethanol / Q000378:metabolism / Q000494:pharmacology; D020404:Glycerophospholipids; D006098:Granulocytes / Q000378:metabolism*; D018922:HL-60 Cells; D006801:Humans; D008970:Molecular Weight; D019301:Oleic Acid / Q000378:metabolism; D010712:Phosphatidic Acids / Q000378:metabolism* / Q000494:pharmacology; D010739:Phospholipase D / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine / Q000378:metabolism*; D011433:Propranolol / Q000494:pharmacology; D011506:Proteins / Q000378:metabolism; D013302:Streptomyces / Q000201:enzymology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000440:Butanols; D020404:Glycerophospholipids; D010712:Phosphatidic Acids; D011506:Proteins; C068564:phosphatidylbutanol; C051521:phosphatidylethanol; D019000:Phosphotyrosine; D019301:Oleic Acid; D000431:Ethanol; D011433:Propranolol; D010739:Phospholipase D; D013755:Tetradecanoylphorbol Acetate