Organic hydroperoxide-induced activation of liver microsomal glutathione S-transferase of rats in vitro.

The effect of t-butyl hydroperoxide (t-BuOOH), cumene hydroperoxide (CuOOH) or linoleic acid hydroperoxide (linoleic-OOH) on liver microsomal glutathione S-transferase of rats was studied in vitro. When microsomes were incubated with either 100 microM t-BuOOH or 25 microM CuOOH, glutathione S-transferase activity was increased 1.5-fold; activity was further increased to 2.2-fold in the presence of small amounts of glutathione. The same amounts of dithiothreitol or cysteine did not enhance the t-BuOOH or CuOOH-induced increase in transferase activity. The transferase activity was also increased 1.4-fold by 10 microM linoleic-OOH plus 1 microM glutathione. The increase in microsomal glutathione S-transferase activity after treatment of microsomes with t-BuOOH in the presence of glutathione was completely reversed by addition of dithiothreitol, whereas the activation of the transferase caused by t-BuOOH in the absence of glutathione was not reversed. Although microsomal glutathione S-transferase also possesses glutathione peroxidase activity, only transferase activity was increased by t-BuOOH in either the presence or absence of glutathione. These data indicate that microsomal glutathione S-transferase is activated by organic hydroperoxides in either the absence or presence of small amounts of glutathione, suggesting an activation of the transferase by thiol oxidation of the cysteine residue.