Aquaculture Studies 2017, Vol 17, Num, 4     (Pages: 385-394)

In vitro Inhibition Effects of Some Metal Ions on Glutathione Reductase Purified from Capoeta trutta Kidney

Muammer KIRICI 1 , Mahinur KIRICI 2 , Muhammed ATAMANALP 3

1 Bingöl University, Faculty of Agriculture, Department of Fisheries, Bingöl, Turkey
2 Bingöl University, Faculty of Arts and Science, Department of Chemistry, Bingöl, Turkey
3 Atatürk University, Faculty of Fisheries, Department of Aquaculture, Erzurum, Turkey
DOI : 10.17693/yunusae.v17i31121.337640 Viewed : 838 - Downloaded : 320 Glutathione reductase (EC: 1.8.1.7; GR) was purified from the kidney of the teleost fish Capoeta trutta. The purification procedure consisted of three steps: preparation of homogenate, ammonium sulfate fractionation and affinity chromatography on 2`,5`-ADP Sepharose 4B. The enzyme was purified 794-fold with a yield of 35.4% and a specific activity of 11.91 U/mg proteins. In order to control enzyme purity, SDS-PAGE was done and showed a single band for enzyme. A single band was obtained approximately at 55 kDa. In addition, inhibitory effects of metal ions (Ag+, Co+2, Ni+2, Cu+2 and Zn+2) on fish kidney glutathione reductase were investigated. Ki constants and IC50 values for metal ions were determined by Lineweaver–Burk graphs and plotting activity % vs. [I], respectively. IC50 values were 0.00078, 0.622, 0.722, 0.073 and 0.519 mM, and Ki constants were 0.000394 ± 0.0002, 0.235 ± 0.027, 0.279 ± 0.048, 0.026 ± 0.008 and 0.382 ± 0.024 mM for Ag+, Co+2, Ni+2, Cu+2 and Zn+2, respectively. From these results, we showed that Ag+ is the most potent inhibitor of glutathione reductase enzyme. Keywords : Glutathione reductase,Capoeta trutta,kidney,inhibitor,purification