从火鸡肝中纯化和表征谷胱甘肽 S-转移酶及几种金属离子对酶活性的抑制作用。
Purification and characterization of glutathione S-transferase from turkey liver and inhibition effects of some metal ions on enzyme activity.
机构信息
Science Faculty, Department of Chemistry, Atatürk University, 25240 Erzurum, Turkey.
出版信息
Environ Toxicol Pharmacol. 2012 Nov;34(3):888-94. doi: 10.1016/j.etap.2012.08.010. Epub 2012 Aug 31.
The glutathione S-transferases (EC 2.5.1.18) were purified and characterized from turkey liver for the first time. The enzyme was purified 252.7-fold with a yield of 45%, with a specific activity of 164.31 U/mg from turkey liver. The purity of the enzyme was determined by SDS-PAGE and showed two bands nearly 26 kDa and 24 kDa on the gel. The native molecular mass of the enzyme was found to be approximately 53 kDa by Sephadex G-100 gel filtration chromatography. Optimal pH, stable pH, optimal temperature, optimum ionic strength, K(m) and V(max) values for GSH and CDNB were also determined for the enzyme as 7.3, 8.5, 50 °C, 600 mM, 0.154 mM, 0.380 mM, 1.803 EU/ml, and 2.125 EU/ml, respectively. Additionally, inhibitory effects of metal ions (Cu(2+), Hg(2+), Fe(2+), Zn(2+), Ag(+), Mg(2+), Ni(2+), and Mn(2+)) were examined the enzyme's activity in vitro by performing Lineweaver-Burk graphs and plotting activity% vs., respectively.
谷胱甘肽 S-转移酶（EC 2.5.1.18）首次从火鸡肝脏中被分离和鉴定。该酶经 252.7 倍纯化，收率为 45%，比活为 164.31 U/mg，从火鸡肝脏中获得。酶的纯度通过 SDS-PAGE 确定，在凝胶上显示出两条带，分别约为 26 kDa 和 24 kDa。通过 Sephadex G-100 凝胶过滤层析发现该酶的天然分子量约为 53 kDa。还确定了该酶对 GSH 和 CDNB 的最适 pH、稳定 pH、最适温度、最佳离子强度、K(m)和 V(max)值，分别为 7.3、8.5、50°C、600 mM、0.154 mM、0.380 mM、1.803 EU/ml 和 2.125 EU/ml。此外，通过进行 Lineweaver-Burk 图谱并分别绘制活性%与各金属离子（Cu(2+)、Hg(2+)、Fe(2+)、Zn(2+)、Ag(+)、Mg(2+)、Ni(2+)和 Mn(2+)）对酶活性的抑制效应，在体外研究了金属离子对该酶活性的抑制作用。
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