Investigation of the Inhibition Effects of Some Metal Ions on Glutathione Reductase Enzyme From Japanese Quail (Coturnix coturnix japonica) Liver

Abstract

In the present study, the inhibition effect of some heavy metal ions on glutathione reductase (GR) enzyme which has important functions in metabolism, was investigated. For this purpose, GR enzyme was purified with 19.33 EU/mg specific activity, a yield of 14.06% and 128.8-fold from quail liver by using ammonium sulphate precipitation and 2ʹ, 5ʹ-ADP Sepharose 4B affinity chromatography. To check the purity of GR enzyme, sodium dodecyl sulfate polyacrilamide gel electrophoresis (SDS-PAGE) was performed and single band was observed. The activity of the GR enzyme was measured spectrophotometrically at 340 nm according to the method of Carlberg and Mannervik. Then, the inhibitory effects of different heavy metal ions (Co 2+, Zn 2+, Pb 2+, Fe 2+, Cd 2+, Al 3+ and Fe 3+) on the activity of GR enzyme were examined under in vitro conditions. For the GR enzyme from quail liver tissue, heavy metal concentrations (IC50) were obtained in which 50% of GR enzyme activity was inhibited in vitro conditions. Finally, Ki values for these metal ions were calculated from the Lineweaver-Burk plots.