Properties of a flavoprotein sulfhydryl oxidase from rat seminal vesicle secretion.
Rat seminal vesicle secretion is a rich source of a flavoprotein oxidase that acts upon sulfhydryl compounds. The enzyme was obtained in homogeneous form as previously described [Ostrowski, M. C., Kistler, W. S., & Williams-Ashman, H. G. (1979) Biochem. Biophy. Res. Commun. 87, 171-176] and characterized with respect to prosthetic group, size, reaction stoichiometry, and substrate specificity. On the basis of its behavior during zone sedimentation, gel filtration, and electrophoresis in the presence of sodium dodecyl sulfate, it appears to be a monomeric enzyme of about 66 000 daltons. Acid denaturation liberates 1 mol of flavin adenine dinucleotide (FAD) per mol of enzyme. The reaction catalyzed was shown to be 2RSH + O2 leads to H2O2. Superoxide formation could be demonstrated. Unlike many flavoprotein oxidases, the enzyme failed to form a bleached complex with sulfite. The enzyme accepts a variety of small sulfhydryl compounds as substrates, including glutathione, cysteine, dithiothreitol, and 2-mercaptoethanol. Michaelis-Menten kinetics were obtained with these substrates providing disulfide contamination was initially eliminated by treating thiols with borohydride. The KM for glutathione was 4.4 mM with a Vmax estimated as 660 mumol per min per mg of protein. The enzyme was capable of markedly enhancing the rate of renaturation of fully reduced ribonuclease. The physiological function of the enzyme is not yet clear, though several possibilities are discussed.
D000818:Animals; D005182:Flavin-Adenine Dinucleotide / Q000032:analysis / Q000494:pharmacology; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D010088:Oxidoreductases / Q000378:metabolism*; D010101:Oxygen Consumption; D011489:Protein Denaturation; D051381:Rats; D012669:Seminal Vesicles / Q000201:enzymology*; D013053:Spectrophotometry; D013379:Substrate Specificity; D050862:Oxidoreductases Acting on Sulfur Group Donors
D005182:Flavin-Adenine Dinucleotide; D010088:Oxidoreductases; D050862:Oxidoreductases Acting on Sulfur Group Donors