title: Superoxide peroxidase activity of ovoperoxidase, the cross-linking enzyme of fertilization.
abstract: Ovoperoxidase, an enzyme secreted by the eggs of the sea urchin Stronglycocentrotus purpuratus upon activation, catalyzes the formation of dityrosine residues in the fertilization envelope. This cross-linking reaction requires extracellular H2O2, which is produced by the egg during the cyanide-insensitive "respiratory burst" of fertilization. While investigating the possibility that the sea urchin oxidase might generate O2- as a precursor to H2O2, we discovered that ovoperoxidase possessed O2- degrading activity. Ovoperoxidase catalyzed the breakdown of O2- in a reaction that was sensitive to inhibition by catalase, indicating a requirement for H2O2. High concentrations of either O2- or H2O2 inhibited the O2- degrading activity of ovoperoxidase, as did the peroxidase inhibitors aminotriazole, azide, and phenylhydrazine. When ovoperoxidase was heated at 56 degrees C, it lost O2- degrading activity in parallel with peroxidase activity. In contrast, the copper-chelating agent diethyldithiocarbamate, which completely inactivated CuZn superoxide dismutase, failed to affect ovoperoxidase. The requirement for H2O2 and the inhibition by aminotriazole, azide, and phenylhydrazine support the hypothesis that ovoperoxidase catalyzes the breakdown of O2- by a peroxidative mechanism. Ovoperoxidase may play a role in protecting the developing embryo from oxidants derived from O2-.
authors.affiliation: Department of Medicine, University of Washington, Seattle 98195.
authors.forename: J W
authors.initials: JW
authors.lastname: Heinecke
authors.forename: B M
authors.initials: BM
authors.lastname: Shapiro
mesh_terms: D000640:Amitrole / Q000494:pharmacology; D000818:Animals; D001386:Azides / Q000494:pharmacology; D002374:Catalase / Q000494:pharmacology; D003574:Cytochrome c Group / Q000378:metabolism; D005306:Fertilization; D006735:Horseradish Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D006358:Hot Temperature; D006861:Hydrogen Peroxide / Q000378:metabolism / Q000494:pharmacology; D007784:Lactoperoxidase / Q000037:antagonists & inhibitors / Q000378:metabolism; D009580:Nitroblue Tetrazolium / Q000378:metabolism; D010063:Ovum / Q000201:enzymology*; D010544:Peroxidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010659:Phenylhydrazines / Q000494:pharmacology; D012617:Sea Urchins; D013482:Superoxide Dismutase / Q000378:metabolism*; D013481:Superoxides / Q000378:metabolism; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism; D014969:Xanthine Oxidase / Q000037:antagonists & inhibitors / Q000378:metabolism
chemical_list: D001386:Azides; D003574:Cytochrome c Group; D010659:Phenylhydrazines; C030299:phenylhydrazine; D013481:Superoxides; D009580:Nitroblue Tetrazolium; D014443:Tyrosine; D006861:Hydrogen Peroxide; C007543:dityrosine; D006735:Horseradish Peroxidase; D007784:Lactoperoxidase; D010544:Peroxidases; D002374:Catalase; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D000640:Amitrole