Association of sulfhydryl oxidase and xanthine oxidase in bovine mammary tissue.
The presence of antigenically active xanthine oxidase was indicated in various relatively purified preparations of sulfhydryl oxidase obtained from bovine milk. Evidence for formation of a complex of the two enzymes was obtained by double immunodiffusion. Furthermore, sodium dodecylsulfate-gel electrophoresis of sulfhydryl oxidase and xanthine oxidase model mixtures indicated that high molecular weight species were present that reacted with both antisulfhydryl oxidase and antixanthine oxidase. Similar gel electrophoretic patterns visualized by protein-dye binding methods revealed a distinct band (greater than 200 kdalton) was formed upon incubation of mixtures of the two enzymes, the presence of which was unaffected by reduction of protein disulfide bonds. Immunofluorescent staining techniques showed both enzymes in the apical plasma membrane. Because sulfhydryl oxidase previously has been shown to catalyze conversion of the dehydrogenase form of xanthine oxidase to the oxidase form, this conversion may occur when xanthine oxidase contacts sulfhydryl oxidase in the apical plasma membrane. This conversion and the resulting potential for production of active oxygen species could be significant to membranotropic processes, such as fat globule secretion, and to the oxidative stability of the milk fat globule membrane.
D000818:Animals; D002417:Cattle; D004586:Electrophoresis; D005260:Female; D008321:Mammary Glands, Animal / Q000032:analysis / Q000201:enzymology*; D008892:Milk / Q000201:enzymology*; D010088:Oxidoreductases / Q000032:analysis / Q000378:metabolism*; D014969:Xanthine Oxidase / Q000032:analysis / Q000378:metabolism*; D050862:Oxidoreductases Acting on Sulfur Group Donors
D010088:Oxidoreductases; D014969:Xanthine Oxidase; D050862:Oxidoreductases Acting on Sulfur Group Donors