Adhesion-dependent protein tyrosine phosphorylation in neutrophils treated with tumor necrosis factor.
Human neutrophils (PMN) respond to tumor necrosis factor (TNF) by releasing their granules, reorganizing their cytoskeleton, and massively secreting hydrogen peroxide. This response is dependent on adhesion to extracellular matrix proteins and expression of CD11b/CD18 integrins (Nathan, C., S. Srimal, C. Farber, E. Sanchez, L. Kabbash, A. Asch, J. Gailit, and S. D. Wright. 1989. J. Cell Biol. 109:1341-1349). We investigated the role of tyrosine phosphorylation in the response of PMN to TNF. PMN adherent to protein-coated surfaces but not suspended PMN showed tyrosine phosphorylation of several proteins (approximately 150, approximately 115, approximately 75, and approximately 65 kD) in response to TNF. Tyrosine phosphorylation was evident 5 min after addition of TNF and lasted at least 2 h. The tyrosine kinase inhibitors K252a, genistein and ST638 suppressed tyrosine phosphorylation and blocked hydrogen peroxide production in a reversible manner at low concentrations. Tyrosine kinase inhibitors also blocked the spreading of PMN in response to TNF. Dihydrocytochalasin B did not inhibit tyrosine phosphorylation, but in its presence phosphorylation was rapidly reversed. By immunocytochemistry, the majority of tyrosine phosphoproteins were localized to focal adhesions. Thus TNF-induced tyrosine phosphorylation depends on adhesion of PMN to extracellular matrix proteins, and participates in the transduction of the signals that direct the cells to spread on a biological surface and undergo a respiratory burst.
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Beatrice and Samuel A. Seaver Laboratory, Department of Cell Biology/Anatomy, Cornell University Medical College, New York 10021.
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Nathan
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D002227:Carbazoles / Q000494:pharmacology*; D002448:Cell Adhesion / Q000187:drug effects*; D002934:Cinnamates / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D019833:Genistein; D006801:Humans; D006861:Hydrogen Peroxide / Q000097:blood; D066298:In Vitro Techniques; D026121:Indole Alkaloids; D007529:Isoflavones / Q000494:pharmacology*; D007700:Kinetics; D008970:Molecular Weight; D009504:Neutrophils / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D010750:Phosphoproteins / Q000302:isolation & purification / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000097:blood*; D011994:Recombinant Proteins / Q000494:pharmacology; D013440:Sulfides / Q000494:pharmacology*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D002227:Carbazoles; D002934:Cinnamates; D016326:Extracellular Matrix Proteins; D026121:Indole Alkaloids; D007529:Isoflavones; D010750:Phosphoproteins; D011994:Recombinant Proteins; D013440:Sulfides; D014409:Tumor Necrosis Factor-alpha; C060747:ST 638; C049985:staurosporine aglycone; D006861:Hydrogen Peroxide; D019833:Genistein; D011505:Protein-Tyrosine Kinases