Purification of glutathione S-transferase isoenzymes from tumour and nontumour human stomach and inhibitory effects of some heavy metals on enzymes activities.
In this study, glutathione S-transferase (GST) enzyme was purified from nontumour and tumour human gastric tissue and in vitro effects of heavy metals on the enzyme were examined. GST was purified 3089 fold with a specific activity of 20 U/mg and a yield of 78% from gastric tumour tissue; and 1185 fold with a specific activity of 5.69 U/mg and a yield of 50% from nontumour tissue by using glutathione-agarose affinity column, respectively. Enzyme purity was verified by SDS-PAGE and subunit molecular mass was calculated around 26 kDa. The molecular weight of the enzyme was calculated as 52 kDa by using Sephadex G-75 gel filtration column. Then, inhibitory effects of metal ions on the enzymes were investigated. Mg(2+) and Cd(2+) had inhibitory effect on the enzymes activities. Other kinetic properties of the enzymes were also determined.
Agrı Ibrahim Cecen University , Vocational Training School, Agrı , Turkey.
Ramazan
R
Demirdag
Emrah
E
Yerlikaya
Omer Irfan
OI
Kufrevioglu
Cemal
C
Gundogdu
D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D019216:Metals, Heavy / Q000494:pharmacology*; D013270:Stomach / Q000201:enzymology*; D013274:Stomach Neoplasms / Q000201:enzymology*; D013329:Structure-Activity Relationship
D007527:Isoenzymes; D019216:Metals, Heavy; D005982:Glutathione Transferase