Purification and characterization of glutaredoxin from Cryptococcus neoformans

Glutaredoxin, also known as thioltransferase, was purified from Cryptococcus neoformans by procedures including DEAE-cellulose ion exchange chromatography, Q-Sepharose ion-exchange chromatography, and gel filtration on Sephadex G-50. Its purity was confirmed by SDS-polyacrylamide gel electrophoresis...

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Bibliographic Details
Published in:Molecules and cells 1997-10, Vol.7 (5), p.655-660
Main Authors: Sa, J.H, Kim, K.H, Lim, C.J. (Kangwon National University, Chunchon (Korea Republic). College of Natural Sciences)
Format: Article
Language:English
Subjects:
BIOLOGIA MOLECULAR
BIOLOGIE MOLECULAIRE
CHAMPIGNON
CHROMATOGRAPHIE
CHROMATOGRAPHY
CROMATOGRAFIA
CRYPTOCOCCUS (CHAMPIGNON)
CRYPTOCOCCUS (FUNGI)
CRYPTOCOCCUS (HONGO)
Cryptococcus neoformans - enzymology
Electrophoresis, Polyacrylamide Gel
Enzyme Stability
Fungal Proteins - chemistry
Fungal Proteins - isolation & purification
Fungal Proteins - metabolism
FUNGI
Glutaredoxins
HONGOS
Hot Temperature
Isoelectric Point
Kinetics
MOLECULAR BIOLOGY
MOLECULAR WEIGHT
Oxidoreductases - chemistry
Oxidoreductases - isolation & purification
Oxidoreductases - metabolism
PESO MOLECULAR
POIDS MOLECULAIRE
Protein Disulfide Reductase (Glutathione)
Proteins - chemistry
Proteins - isolation & purification
Proteins - metabolism
Substrate Specificity
Sulfhydryl Compounds
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Description
Summary:Glutaredoxin, also known as thioltransferase, was purified from Cryptococcus neoformans by procedures including DEAE-cellulose ion exchange chromatography, Q-Sepharose ion-exchange chromatography, and gel filtration on Sephadex G-50. Its purity was confirmed by SDS-polyacrylamide gel electrophoresis and its molecular weight was estimated to be 12,000 Da. The purified enzyme has a K(m) value of 1.03 mM with 2-hydroxyethyl disulfide as a substrate. The enzyme also utilizes L-sulfocysteine, L-cystine, and bovine serum albumin as substrates in the presence of reduced glutathione. The enzyme has K(m) values of 0.34-2.50 mM for these substrates. It was greatly activated by thiol compounds such as reduced glutathione, dithiothreitol, L-cysteine and beta-mercaptoethanol. It is partially inactivated at 60 degrees C or higher temperatures. It plays an important role in thiol-disulfide exchange in Cryptococcus neoformans.
ISSN:1016-8478
DOI:10.1016/S1016-8478(23)13354-1