A single step method for the separation of rat liver cytosolic glutathione S-transferase isozymes

A simple, single step chromatographic method was developed to separate the liver cytosolic glutathione S-transferase (GSH-S-t) isozymes from each other and from the bulk of the cytosolic protein. Five peaks of GSH-S-t activity, tested with 1-chloro-2,4-dinitrobenzene (CDNB) as a substrate, were eluted. By comparison of the activities with CDNB and the other substrate 3,4-dichloronitrobenzene (DCNB) the five peaks could be identified as GSH-S-t isozymes C, B, A, and AA, being GSH-S-t isozyme C eluted in two different peaks. The method was used to detect a decrease of specific GSH-S-t isozymes in the cytosol of rats intoxicated with carbon tetrachloride, as compared with control rats.