Respiratory burst and tyrosine phosphorylation by vanadate.
We studied involvement of tyrosine-phosphorylated proteins in activation of NADPH oxidase in guinea pig neutrophils. Pervanadate, which is the oxidized form of orthovanadate, induced O2- production and protein tyrosine phosphorylation in neutrophils. O2- production induced by pervanadate was more sensitive to the tyrosine kinase-specific inhibitor, ST-638, as compared with the production induced by PMA. On the other hand, staurosporine more selectively inhibited PMA-induced O2- production than pervanadate-induced production. These results indicate that tyrosine kinase, not protein kinase C, is involved in pervanadate-induced O2- production. The tyrosine-phosphorylated proteins were detected in both the cytosol and membrane fractions prepared from pervanadate-induced neutrophils. In order to examine if tyrosine residues of some components of NADPH oxidase were directly phosphorylated, tyrosine-phosphorylated proteins were removed from solubilized membranes prepared from the pervanadate-stimulated neutrophils by immunoprecipitation with an anti-phosphotyrosine anti-body. NADPH oxidase activity in the solubilized membranes was not decreased by the treatment. These findings suggest that the components of NADPH oxidase are not tyrosine-phosphorylated by pervanadate treatment, that tyrosine phosphorylation may be involved in the signal transduction pathway of NADPH oxidase activation by pervanadate, and that this pathway is independent of the activation by protein kinase C.
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006168:Guinea Pigs; D008565:Membrane Proteins / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D019000:Phosphotyrosine / Q000378:metabolism*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D016897:Respiratory Burst / Q000187:drug effects*; D015398:Signal Transduction; D019311:Staurosporine; D013481:Superoxides / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology*
D000470:Alkaloids; D004791:Enzyme Inhibitors; D008565:Membrane Proteins; D010750:Phosphoproteins; C076715:pervanadate; D013481:Superoxides; D019000:Phosphotyrosine; D014638:Vanadates; D011493:Protein Kinase C; D019311:Staurosporine