Enzymatic determination of membrane lipid peroxidation

Abstract

The recently purified “phospholipid hydroperoxide glutathione peroxidase” has been used to measure the membrane hydroperoxides formed during lipid peroxidation that are not substrates for the “classical” glutathione peroxidase. A spectrophotometric test in the presence of glutathione, glutathione reductase and NADPH has been used. The peroxidized membranes were added directly to the reaction mixture and the reaction was started by the addition of the enzyme. Triton X-100 exerted a stimulatory effect. Phospholipid hydroperoxide glutathione peroxidase allows a rapid, sensitive, accurate and specific determination of membrane hydroperoxides, the most quantitative index of lipid peroxidation. Glutathione peroxidase can be used in the same test to measure other hydroperoxides such as the cumene hydroperoxide used to induce the peroxidation.