Superoxide-dependent hydroxylation by myeloperoxidase.
When stimulated, neutrophils undergo a respiratory burst converting oxygen to superoxide. Although superoxide is critical for microbial killing by phagocytic cells, the precise role it plays has yet to be established. It has been proposed to optimize their production of hypochlorous acid and to be required for the generation of hydroxyl radicals. Superoxide is also involved in the hydroxylation of salicylate by neutrophils. However, the mechanism of this reaction is unknown. We found that neutrophils stimulated with opsonized zymosan hydroxylated salicylate to produce mainly 2,5-dihydroxybenzoate. Its formation was dependent on superoxide and a heme protein but was independent of hydrogen peroxide and hydroxyl radicals. Production of 2,5-dihydroxybenzoate was enhanced by methionine, which scavenges hypochlorous acid. Neutrophils from an individual with myeloperoxidase deficiency hydroxylated salicylate at only 13% of the level of control cells. Purified human myeloperoxidase and xanthine oxidase plus hypoxanthine hydroxylated salicylate to produce 2,5-dihydroxybenzoate. As with neutrophils, the reaction required superoxide but not hydrogen peroxide and was unaffected by hydroxyl radical scavengers. Thus, myeloperoxidase catalyzes superoxide-dependent hydroxylation. This newly recognized reaction may be relevant to the in vivo functions of superoxide and myeloperoxidase.
Kettle
A J
AJ
Department of Pathology, Christchurch School of Medicine, New Zealand.
Winterbourn
C C
CC
D006801:Humans; D062385:Hydroxybenzoates / Q000378:metabolism; D006900:Hydroxylation; D006997:Hypochlorous Acid / Q000737:chemistry; D066298:In Vitro Techniques; D009504:Neutrophils / Q000378:metabolism*; D009195:Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012459:Salicylates / Q000378:metabolism*; D013057:Spectrum Analysis; D013481:Superoxides / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000378:metabolism
D062385:Hydroxybenzoates; D012459:Salicylates; D013481:Superoxides; D006997:Hypochlorous Acid; D009195:Peroxidase; D014969:Xanthine Oxidase