title: Glucocerebroside inhibits NADPH oxidase activation in cell-free system.
abstract: We reported earlier that monocytes and macrophages from patients with type I Gaucher disease have a decreased capacity to generate superoxide anion (O(2)(-)) on stimulation with opsonized S. aureus or formyl-methionyl-leucyl-phenylalanine. In this study, various forms of the cell-free assay system were used to probe the hypothesis that glucocerebroside (GC) accumulating in Gaucher patients' phagocytes may interfere with the activation of NADPH oxidase. Xanthine/xanthine oxidase assay was applied to explore the possibility that GC may scavenge O(2)(-). We found that addition of GC to the crude, semirecombinant or fully purified cell-free systems inhibited activation of NADPH oxidase in a concentration-dependent manner. The inhibitory effect of GC could be overcome by increased concentrations of p47(phox) and p67(phox). In contrast, O(2)(-) generation was not decreased by GC added to the assembled, catalytically active enzyme complex. In the xanthine/xanthine oxidase system, GC had no effect on the generation of O(2)(-). These data indicate that assembly of the respiratory burst oxidase of phagocytic cells may be a possible target of the pathologic actions of GC.
authors.affiliation: Department of Physiology, Semmelweis University, Budapest, Hungary.
authors.forename: Patryk
authors.initials: P
authors.lastname: Moskwa
authors.forename: Anita
authors.initials: A
authors.lastname: Palicz
authors.forename: Marie-Hélène
authors.initials: MH
authors.lastname: Paclet
authors.forename: Marie-Claire
authors.initials: MC
authors.lastname: Dagher
authors.forename: Melinda
authors.initials: M
authors.lastname: Erdos
authors.forename: László
authors.initials: L
authors.lastname: Maródi
authors.forename: Erzsébet
authors.initials: E
authors.lastname: Ligeti
mesh_terms: D002474:Cell-Free System; D003001:Cloning, Molecular; D004789:Enzyme Activation / Q000187:drug effects; D005963:Glucosylceramides / Q000494:pharmacology*; D006801:Humans; D007700:Kinetics; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000097:blood* / Q000187:drug effects; D009504:Neutrophils / Q000201:enzymology*; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D013481:Superoxides / Q000097:blood; D014969:Xanthine Oxidase / Q000097:blood
chemical_list: D005963:Glucosylceramides; D011994:Recombinant Proteins; D013481:Superoxides; D014969:Xanthine Oxidase; D019255:NADPH Oxidases