Study on human erythrocyte thioltransferase: comparative characterization with bovine enzyme and its physiological role under oxidative stress.
Thioltransferase, an enzyme which catalyzes the thiol/disulfide exchange reaction in the presence of GSH, was purified to homogeneity on 15% SDS-PAGE from human (36,000-fold purification) and bovine (23,000-fold) erythrocyte hemolysates. These enzymes had similar properties in their monomeric structures (M(r) = 11,000) and broad specificities for substrates ranging from low-molecular disulfides (S-sulfocysteine, cystamine, and cystine) to protein disulfides (trypsin and insulin). They were highly sensitive to SH-reagents (monoiodoacetic acid and mercuric chloride), but were protected from inactivation by the presence of disulfides (GSSG, cystamine, and cystine). Phosphofructokinase and pyruvate kinase that had been inactivated by disulfides were reactivated effectively by the addition of thioltransferase with GSH. In addition, disulfides in membrane proteins of human erythrocytes that have been oxidatively damaged by diamide treatment were reduced to the SH-free form more effectively by incubation with thioltransferase.
Terada
T
T
Laboratory of Biochemistry, Faculty of Pharmaceutical Sciences, Osaka University.
Oshida
T
T
Nishimura
M
M
Maeda
H
H
Hara
T
T
Hosomi
S
S
Mizoguchi
T
T
Nishihara
T
T
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002417:Cattle; D004220:Disulfides / Q000097:blood; D004910:Erythrocyte Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000201:enzymology*; D054477:Glutaredoxins; D006801:Humans; D008565:Membrane Proteins / Q000097:blood; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000037:antagonists & inhibitors / Q000097:blood* / Q000737:chemistry; D011490:Protein Disulfide Reductase (Glutathione); D013045:Species Specificity; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000097:blood
D000596:Amino Acids; D004220:Disulfides; D054477:Glutaredoxins; D008565:Membrane Proteins; D013438:Sulfhydryl Compounds; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione)