Different effects of Triton X-100, deoxycholate, and fatty acids on the kinetics of glutathione peroxidase and phospholipid hydroperoxide glutathione peroxidase.
The effects of Triton X-100, deoxycholate, and fatty acids were studied on the two steps of the ping-pong reaction catalyzed by Se-dependent glutathione peroxidases. The study was carried out by analyzing the single progression curves where the specific glutathione oxidation was monitored using glutathione reductase and NADPH. While the "classic" glutathione peroxidase was inhibited only by Triton, the newly discovered "phospholipid hydroperoxide glutathione peroxidase" was inhibited by deoxycholate and by unsaturated fatty acids. The kinetic analysis showed that in the case of glutathione peroxidase only the interaction of the lipophilic peroxidic substrate was hampered by Triton, indicating that the enzyme is not active at the interface. Phospholipid hydroperoxide glutathione peroxidase activity measured with linoleic acid hydroperoxide as substrate, on the other hand, was not stimulated by the Triton concentrations which have been shown to stimulate the activity on phospholipid hydroperoxides. Furthermore a slight inhibition was apparent at high Triton concentrations and the effect could be attributed to a surface dilution of the substrate. Deoxycholate and unsaturated fatty acids were not inhibitory on glutathione peroxidase but inhibited both steps of the peroxidic reaction of phospholipid hydroperoxide glutathione peroxidase, in the presence of either amphiphilic or hydrophilic substrates. This inhibition pattern suggests an interaction of anionic detergents with the active site of this enzyme. These results are in agreement with the different roles played by these peroxidases in the control of lipid peroxide concentrations in the cells. While glutathione peroxidase reduces the peroxides in the water phase (mainly hydrogen peroxide), the new peroxidase reduces the amphyphilic peroxides, possibly at the water-lipid interface.
D001665:Binding Sites; D002384:Catalysis; D003840:Deoxycholic Acid / Q000494:pharmacology*; D005227:Fatty Acids / Q000494:pharmacology*; D005979:Glutathione Peroxidase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D007700:Kinetics; D008041:Linoleic Acids / Q000378:metabolism; D008054:Lipid Peroxides / Q000378:metabolism; D017830:Octoxynol; D019301:Oleic Acid; D009829:Oleic Acids / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology*; D013379:Substrate Specificity
D005227:Fatty Acids; D008041:Linoleic Acids; D008054:Lipid Peroxides; D009829:Oleic Acids; D003840:Deoxycholic Acid; C008285:linoleic acid hydroperoxide; D019301:Oleic Acid; D011092:Polyethylene Glycols; D017830:Octoxynol; D005979:Glutathione Peroxidase