Tyrosine-specific protein phosphorylation during activation of human neutrophils.
The activation of human neutrophils by a variety of receptor-dependent and receptor-independent agonists induces the phosphorylation of a large number of proteins. Since we have previously shown that human neutrophils have at least two distinct tyrosine kinase activities, we examined protein tyrosine phosphorylation in human neutrophils stimulated with a variety of agonists. Using a monoclonal antibody specific for phosphotyrosine, the present study shows that the chemotactic peptides FMLP and leukotriene B4, the phorbol ester phorbol myristate acetate (PMA), and the calcium ionophore A23187 induce an increase in tyrosine phosphorylation of a number of neutrophil proteins. This increased protein tyrosine phosphorylation was dependent on the concentration of the agonist, as well as on the time of exposure to the agonist. Fractionation experiments showed that both a 150,000 g cytosolic and a particulate preparation showed increases in protein tyrosine phosphorylation with stimulation by FMLP or PMA, and showed that the pattern of protein tyrosine phosphorylation was slightly different in the FMLP- and PMA-stimulated cells. These data indicate that protein tyrosine phosphorylation is an early event in the activation of human neutrophils by a variety of receptor-dependent and receptor-independent agonists.
Department of Pediatrics, University of Alabama, Birmingham.
R L
RL
Berkow
R W
RW
Dodson
D015153:Blotting, Western; D000001:Calcimycin / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D007975:Leukotriene B4 / Q000494:pharmacology; D008970:Molecular Weight; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D019000:Phosphotyrosine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
D010750:Phosphoproteins; D007975:Leukotriene B4; D019000:Phosphotyrosine; D000001:Calcimycin; D014443:Tyrosine; D009240:N-Formylmethionine Leucyl-Phenylalanine; D013755:Tetradecanoylphorbol Acetate