Evidence for identifying fatty acids in rat liver glutathione S-transferase and its possible involvement in the...

A rat liver glutathione S-transferase isozyme (GST 3-3) has been purified to apparent electrophoretic homogeneity by procedures including Sephadex G-100, S-hexylglutathione-linked Sepharose, and CM-cellulose column chromatographies. The present study provides evidence for the existence of endogenous fatty acids in the purified GST 3-3 by gas-liquid chromatographic and mass-spectrometric analyses. The liver GST isozyme associated long chain fatty acids such as 14:0, 16:0, 18:0, and 18:1 and the molar ratio of fatty acids to the protein was estimated to be around 3.2. When the enzyme preparation was freed of fatty acids by a mild delipidization technique using Lipidex, the GST activity was significantly decreased. Computer analysis of the circular dichroism spectra revealed that rat liver GST 3-3 contained approximately 49% alpha-helix and 24% random coil. By delipidizing, the enzyme's alpha-helix was decreased to 4% and the random coil was in turn increased to 62%. The enzymatic and structural properties of the delipidized enzyme, however, could be restored to nearly the original levels by recombining with the fatty acids. These findings suggest that weakly bound fatty acids are responsible for the functional capacity of the GST by virtue of changing the protein conformation.