Attempt to differentiate between individual glutathione peroxidases in biological samples

Abstract

Routine determinations of glutathione peroxidases (GPx) still suffer from poor standardization and usually from lack of specification. The different types of glutathione peroxidases present in cellular homogenates may be differentially estimated by their distinct substrate specificities. Complex lipid hydroperoxides required for this approach, however, are not generally available nor easily standardized due to their molecular heterogeneity and tendency to form micelles. We therefore developed a simple procedure for the differential estimation of the major cellular types of GPx, the cytosolic GPx (cGPx) and the phospholipid hydroperoxide glutathione peroxidase (PHGPx) taking advantage of the peculiar susceptibility of PHGPx to deoxycholate. It proved to reliably determine the activities of both purified cGPx and PHGPx, in mixtures thereof, and in homogenates of tissue samples (e.g., testes), and some (e.g. ECV 304) but not all (e.g. THP-1) cultured cell lines. The method allows the differential estimation of cGPx and PHGPx, if the samples do not contain further types of GPx.