Hck is activated by opsonized zymosan and A23187 in distinct subcellular fractions of human granulocytes.
Regulation of neutrophil responses is known to involve tyrosine phosphorylation. Hck, a major neutrophil protein-tyrosine kinase, becomes expressed during differentiation of human promyelocytic NB4 cells into neutrophil-like cells. Hck is mainly localized in a secretory granule-enriched cell fraction, but it is also present in a granule-free membrane fraction and the cytosol. Hck is rapidly and transiently activated upon stimulation of differentiated NB4 cells or human neutrophils with serum-opsonized zymosan or the calcium ionophore A23187, but not by phorbol 12-myristate 13-acetate. In NB4 cells, Hck is also weakly activated by fMet-Leu-Phe. Cell fractionation showed that opsonized zymosan and A23187 induce Hck activation in distinct subcellular fractions. Both stimuli activate Hck in the secretory granule-enriched fraction, but only A23187 activates the kinase in the granule-free membrane fraction. Our results suggest that Hck might regulate early signal transduction events induced by opsonized zymosan and A23187, and that the different subcellular fractions of Hck might serve discrete functions, one of which could be regulation of the degranulation response.
Welch
H
H
Institut de Pharmacologie et de Biologie Structurale, CNRS, UPR 9062, Toulouse, France.
Maridonneau-Parini
I
I
D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D002451:Cell Compartmentation; D015550:Cell Degranulation; D002454:Cell Differentiation; D002478:Cells, Cultured; D003594:Cytoplasmic Granules / Q000201:enzymology; D004789:Enzyme Activation; D006801:Humans; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D019255:NADPH Oxidases / Q000378:metabolism; D009504:Neutrophils / Q000201:enzymology* / Q000648:ultrastructure; D009895:Opsonin Proteins / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D051572:Proto-Oncogene Proteins c-hck; D013347:Subcellular Fractions / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology; D015054:Zymosan
D009895:Opsonin Proteins; D011518:Proto-Oncogene Proteins; D000001:Calcimycin; D014212:Tretinoin; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015054:Zymosan; D019255:NADPH Oxidases; D011505:Protein-Tyrosine Kinases; C496631:HCK protein, human; D051572:Proto-Oncogene Proteins c-hck; D013755:Tetradecanoylphorbol Acetate; D002118:Calcium