title: Signal transduction mechanisms of C1q-mediated superoxide production. Evidence for the involvement of temporally distinct staurosporine-insensitive and sensitive pathways.
abstract: C1q, a plasma glycoprotein and the recognition component of the classical complement pathway, interacts with specific cells of the immune system resulting in the enhancement of cell function. For example, interaction of C1q with its cell-surface receptor on neutrophils induces the activation of the respiratory burst, a finding previously documented using a chemiluminescent assay to detect oxygen radical formation. In an alternative approach we have now used a modified cytochrome c reduction assay to characterize C1q-mediated production of superoxide anion (O2-) in more detail. C1q coated to microtiter wells induced O2- release, which occurred microtiter wells induced O2- release, which occurred after a lag period of 10 to 20 min, and was then sustained over approximately 1 h. O2- production could be triggered by the purified pepsin-resistant, collagen-like fragment of C1q, but not by mannose-binding protein and pulmonary surfactant protein A, proteins that also contain collagen-like domains. Concentrations of C1q which promoted a vigorous O2- generation did not induce release of neutrophil primary granules and caused little or no secondary granule release. Investigation of the biochemical events mediating C1q stimulated O2- production by neutrophils revealed that the response invoked two biochemical pathways with distinct sensitivities to previously described inhibitors. A role for Ca2+ in initiation of the response was suggested by the inhibitory effect of EGTA, the calmodulin antagonist W7, and the intracellular Ca2+ chelator BAPTA. The protein kinase inhibitor staurosporine did not inhibit the induction of the response, but did block that component of the response occurring after approximately 30 min. Neither phase of C1q-mediated O2- production was inhibited by pertussis toxin, a strong inhibitor of the G-protein-coupled FMLP-mediated response. In summary, C1q-triggered O2- production is relatively unique both in terms of the kinetics of the response and the biochemical pathways evoked. These data support the hypothesis that more than one biochemical pathway induced by ligand-receptor interaction can activate the neutrophil NADPH oxidase.
authors.affiliation: Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
authors.forename: E B
authors.initials: EB
authors.lastname: Goodman
authors.forename: A J
authors.initials: AJ
authors.lastname: Tenner
mesh_terms: D000470:Alkaloids / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D015550:Cell Degranulation / Q000187:drug effects; D002478:Cells, Cultured; D015922:Complement C1q / Q000494:pharmacology*; D005966:Glucuronidase / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D016177:Macrophage-1 Antigen / Q000378:metabolism; D009113:Muramidase / Q000378:metabolism; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000502:physiology*; D016897:Respiratory Burst; D015398:Signal Transduction; D019311:Staurosporine; D013481:Superoxides / Q000378:metabolism*; D013997:Time Factors; D014155:Transcobalamins / Q000378:metabolism
chemical_list: D000470:Alkaloids; D016177:Macrophage-1 Antigen; D014155:Transcobalamins; D013481:Superoxides; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015922:Complement C1q; D009113:Muramidase; D005966:Glucuronidase; D019311:Staurosporine; D002118:Calcium