Thioltransferase from Schizosaccharomyces pombe: purification to homogeneity and some properties.
Two types of thioltransferase were identified in the cytosolic extract of Schizosaccharomyces pombe, a fission yeast. In the present study, the major one of them was purified to homogeneity using chromatography processes such as ion-exchange chromatography and gel filtration. Purification was monitored by the transhydrogenase activity of thioltransferase with 2-hydroxyethyl disulfide as a substrate. Its molecular weight was estimated to be about 14,000 on SDS-polyacrylamide gel electrophoresis. The purified enzyme catalyzes the reduction of various disulfide compounds such as S-sulfocysteine, L-cystine, and insulin. It was also found to contain the reducing activity on non-disulfide substrates such as dehydroascorbic acid and alloxan. Its activity was greatly activated by high concentrations of reduced glutathione. It was found to be very heat-stable as like other thioltransferases. It was characterized on other aspects such as kinetic parameters and optimal reaction conditions.
Kim
H G
HG
Division of Life Sciences, College of Natural Sciences, Kangwon National University, Chuncheon, Korea.
Park
E H
EH
Lim
C J
CJ
D000931:Antidotes / Q000494:pharmacology; D002412:Cations / Q000494:pharmacology; D003545:Cysteine / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004795:Enzyme Stability; D054477:Glutaredoxins; D005978:Glutathione / Q000494:pharmacology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology; D008670:Metals / Q000494:pharmacology; D008970:Molecular Weight; D010088:Oxidoreductases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011490:Protein Disulfide Reductase (Glutathione); D012568:Schizosaccharomyces / Q000187:drug effects / Q000201:enzymology*; D013379:Substrate Specificity; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D000931:Antidotes; D002412:Cations; D054477:Glutaredoxins; D008670:Metals; D013439:Sulfhydryl Reagents; D008623:Mercaptoethanol; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione); D005978:Glutathione; D003545:Cysteine; D004229:Dithiothreitol