Purification and characterization of glutathione-S-transferase from chicken erythrocyte
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2009
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Abstract
The glutathione-s-transferases are a family of multifunctional enzymes involved in the detoxification of electrophilic xenobiotics primarily through conjugation to reduce glutathione. A form of the enzyme, designated GSH-S transferase ρ, was purified chicken erythrocyte by acetone precipitation, ethanol-chloroform treatment, DEAE-Cellulose, Q-Sepharose, Sephadex G-100 chromatography. The molecular weight of GST purified from chicken erythrocyte was estimated as 47,500 Da by gel filtration. The subunit molecular weight of chicken erythrocyte GST as determined by electrophoresis in the presence of sodium dodecyl sulfate was predicted as 24,000 Da. The specific activity was found to be 20.39 U/mg. The km for CDNB calculated from Lineweaver-Burk plot was 0.71 mM. Optimum temperature of maximum GST activity was 28°C for CDNB. The maximal activity of the enzyme was observed at pH 7.5. The activity of purified GST is inhibited by DDT, urea, CDNB, Triton X-100, DTNB. Copyright © 2009 Informa UK Ltd.
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Acetone , Animals , Chickens , Chromatography, DEAE-Cellulose , DDT , Dithionitrobenzoic Acid , Enzyme Activation , Erythrocytes , Glutathione Transferase , Hydrogen-Ion Concentration , Molecular Weight , Octoxynol , Precipitation , Temperature , Urea , Acetone , Biochemistry , Chromatographic analysis , Detoxification , Electrophoresis , Enzyme activity , Ethanol , Gel permeation chromatography , Gelation , Ion exchange , Meats , Molecular weight , Purification , Sodium , Sodium sulfate , System theory , Urea , alcohol , chloroform , diethylaminoethyl cellulose , dodecyl sulfate sodium , glutathione transferase , sephadex , sepharose , Chicken erythrocyte , Chloroform treatments , Gel filtrations , Glutathione , Glutathione-S-transferase , Inhibitors , Lineweaver-Burk plots , Optimum temperatures , Q-sepharose , Sephadex , Sodium dodecyl sulfates , Specific activities , Triton x-100 , acid precipitation , animal cell , article , chromatography , detoxification , electrophoresis , enzyme activity , enzyme analysis , enzyme purification , enzyme specificity , erythrocyte , gel filtration , molecular weight , nonhuman , polyacrylamide gel electrophoresis , prediction , protein family , xenobiotic metabolism , Enzymes