Identification of the dehydroascorbic acid reductase and thioltransferase (Glutaredoxin) activities of bovine erythrocyte glutathione peroxidase.
Bovine erythrocyte glutathione (GSH) peroxidase (GPX, EC 1.11.1.9) was examined for GSH-dependent dehydroascorbate (DHA) reductase (EC 1.8.5.1) and thioltransferase (EC 1.8.4.1) activities. Using the direct assay method for GSH-dependent DHA reductase activity, GPX had a kcat (app) of 140 +/- 9 min-1 and specificity constants (kcat/Km(app)) of 5.74 +/- 0.78 x 10(2) M-1s-1 for DHA and 1.18 +/- 0.17 x 10(3) M-1s-1 for GSH based on the monomer Mr of 22,612. Using the coupled assay method for thioltransferase activity, GPX had a kcat (app) of 186 +/- 9 min-1 and specificity constants (app) of 1. 49 +/- 0.14 x 10(3) M-1s-1 for S-sulfocysteine and 1.51 +/- 0.18 x 10(3) M-1s-1 for GSH based on the GPX monomer molecular weight. GPX has a higher specificity constant for S-sulfocysteine than DHA, and both assay systems gave nearly identical specificity constants for GSH. The DHA reductase and thioltransferase activities of GPX adds to the repertoire of functions of this enzyme as an important protector against cellular oxidative stress.
Washburn
M P
MP
Department of Biochemistry, Michigan State University, East Lansing, Michigan, 48824, USA.
Wells
W W
WW
D000818:Animals; D002417:Cattle; D003545:Cysteine / Q000031:analogs & derivatives / Q000378:metabolism; D003683:Dehydroascorbic Acid / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000201:enzymology*; D054477:Glutaredoxins; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000302:isolation & purification / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology; D008970:Molecular Weight; D010088:Oxidoreductases / Q000378:metabolism*; D011490:Protein Disulfide Reductase (Glutathione); D013552:Swine
D054477:Glutaredoxins; C011119:S-sulphocysteine; D010088:Oxidoreductases; D005979:Glutathione Peroxidase; D011490:Protein Disulfide Reductase (Glutathione); C020666:glutathione dehydrogenase (ascorbate); D005978:Glutathione; D003545:Cysteine; D003683:Dehydroascorbic Acid