Department of Chemistry, Science Faculty, Ataturk University, Erzurum, Turkey.
J Enzyme Inhib Med Chem. 2012 Feb;27(1):18-23. doi: 10.3109/14756366.2011.572879. Epub 2011 Jul 8.
The effects of ketotifen, meloxicam, phenyramidol-HCl and gadopentetic acid on the enzyme activity of GR were studied using human erythrocyte glutathione reductase (GR) enzymes in vitro. The enzyme was purified 209-fold from human erythrocytes in a yield of 19% with 0.31 U/mg. The purification procedure involved the preparation of haemolysate, ammonium sulphate precipitation, 2'',5'-ADP Sepharose 4B affinity chromatography and Sephadex G-200 gel filtration chromatography. Purified enzyme was used in the in vitro studies. In the in vitro studies, IC(50) values and K(i) constants were 0.012 mM and 0.0008 ± 0.00021 mM for ketotifen; 0.029 mM and 0.0061 ± 0.00127 mM for meloxicam; 0.99 mM and 0.4340 ± 0.0890 mM for phenyramidol-HCl; 138 mM and 28.84 ± 4.69 mM for gadopentetic acid, respectively, showing the inhibition effects on the purified enzyme. Phenyramidol-HCl showed competitive inhibition, whereas the others showed non-competitive inhibition.
本研究采用体外人红细胞谷胱甘肽还原酶（GR）研究了酮替芬、美洛昔康、苯异恶唑盐酸盐和钆喷替酸对 GR 酶活性的影响。酶从人红细胞中经 209 倍纯化，产率为 19%，酶活为 0.31U/mg。纯化过程包括血红蛋白的制备、硫酸铵沉淀、2'',5'-ADP Sepharose 4B 亲和层析和 Sephadex G-200 凝胶过滤层析。在体外研究中使用纯化的酶。在体外研究中，酮替芬的 IC50 值和 K(i)常数分别为 0.012mM 和 0.0008±0.00021mM；美洛昔康的 IC50 值和 K(i)常数分别为 0.029mM 和 0.0061±0.00127mM；苯异恶唑盐酸盐的 IC50 值和 K(i)常数分别为 0.99mM 和 0.4340±0.0890mM；钆喷替酸的 IC50 值和 K(i)常数分别为 138mM 和 28.84±4.69mM，均显示对纯化酶的抑制作用。苯异恶唑盐酸盐表现出竞争性抑制，而其他物质表现出非竞争性抑制。
