Purification and characterization of glutathione-s-transferase from chicken erythrocyte.
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 rho, 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 degrees 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.
Aydemir
Tulin
T
Celal Bayar University, Science and Arts Faculty, Chemistry Department, Muradiye-Manisa, Turkey. aydemirt@yahoo.com
Kavrayan
Demet
D
D000096:Acetone; D000818:Animals; D011232:Chemical Precipitation; D002645:Chickens; D002848:Chromatography, DEAE-Cellulose; D003634:DDT; D004228:Dithionitrobenzoic Acid; D004789:Enzyme Activation / Q000187:drug effects* / Q000502:physiology*; D004912:Erythrocytes / Q000201:enzymology*; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D017830:Octoxynol; D013696:Temperature; D014508:Urea
D000096:Acetone; D014508:Urea; D017830:Octoxynol; D004228:Dithionitrobenzoic Acid; D003634:DDT; D005982:Glutathione Transferase