Kuzu Muslum, Aslan Abdulselam, Ahmed Ishtiaq, Comakli Veysel, Demirdag Ramazan, Uzun Naim
Faculty of Pharmacy, University of Ağrı İbrahim Çeçen, 04100, Ağrı, Turkey.
Department of Nutrition and Dietetics, University of Ağrı İbrahim Çeçen, Ağrı, Turkey.
Fish Physiol Biochem. 2016 Apr;42(2):483-91. doi: 10.1007/s10695-015-0153-7. Epub 2015 Dec 16.
Glucose-6-phosphate dehydrogenase (G6PD) and glutathione reductase (GR) are metabolically quite important enzymes. Within this study, these two enzymes were purified for the first time from the gills of Lake Van fish. In the purifying process, ammonium sulfate precipitation and 2',5'-ADP Sepharose 4B affinity column chromatography techniques for glucose-6-phosphate dehydrogenase, temperature degradation and 2',5'-ADP Sepharose 4B affinity column chromatography for glutathione reductase enzyme were used. The control of the enzyme purity and determination of molecular weight were done with sodium dodecyl sulfate polyacrylamide gel electrophoresis. K(M) and V(max) values were determined with Lineweaver-Burk plot. Besides, the effects of some chalcone derivatives on the purified enzymes were analyzed. For the ones showing inhibition effect, % activity-[I] figures were drawn and IC50 values were determined. K(i) value was calculated by using Cheng-Prusoff equation.
葡萄糖-6-磷酸脱氢酶（G6PD）和谷胱甘肽还原酶（GR）是代谢过程中非常重要的酶。在本研究中，首次从凡湖鱼的鳃中纯化出这两种酶。在纯化过程中，葡萄糖-6-磷酸脱氢酶采用硫酸铵沉淀和2',5'-二磷酸腺苷琼脂糖4B亲和柱层析技术，谷胱甘肽还原酶采用温度降解和2',5'-二磷酸腺苷琼脂糖4B亲和柱层析技术。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳对酶的纯度进行检测并测定分子量。用Lineweaver-Burk图测定K(M)和V(max)值。此外，还分析了一些查尔酮衍生物对纯化酶的影响。对于表现出抑制作用的衍生物，绘制了%活性-[I]图并测定了IC50值。通过Cheng-Prusoff方程计算K(i)值。
