Detoxication of base propenals and other alphabeta-unsaturated aldehyde products of radical reactions and lipid peroxidationby human glutathione transferases.

摘要

Radiation and chemical reactions that give rise to free radicals cause the formation of highly cytotoxic base propenals, degradation products of DNA. Human glutathione transferases (GSTs; RX:glutathione R-transferase, EC 2.5.1.18) of classes Alpha, Mu, and Pi were shown to promote the conjugation of glutathione with base propenals and related alkenes. GST P1-1 was particularly active in catalyzing the reactions with the propenal derivatives, and adenine propenal was the substrate giving the highest activity. The catalytic efficiency of GST P1-1 with adenine propenal (kcat/Km = 7.7 x 10(5) M-1.s-1) is the highest so far reported with any substrate for this enzyme. In general, GST A1-1 and GST M1-1, in contrast to GST P1-1, were more active with 4-hydroxyalkenals (products of lipid peroxidation) than with base propenals. The adduct resulting from the Michael addition of glutathione to the alkene function of one of the base propenals (adenine propenal) was identified by mass spectrometry. At the cellular level, GST P1-1 was shown to provide protection against alpha, beta-unsaturated aldehydes. GST P1-1 added to the culture medium of HeLa cells augmented the protective effect of glutathione against the toxicity of adenine propenal and thymine propenal. No protectiveeffect of the enzyme was observed in the presence of the competitive inhibitorS-hexylglutathione. GST P1-1 introduced into Hep G2 cells by electroporation wassimilarly found to increase their resistance to acrolein. The results show thatglutathione transferases may play an important role in cellular detoxication ofelectrophilic alpha, beta-unsaturated carbonyl compounds produced by radicalreactions, lipid peroxidation, ionizing radiation, and drugmetabolism.