Deuterium kinetic isotope effect (DKIE) in copper-induced LDL peroxidation: Interrelated effects of on inhibition and propagation

LDL peroxidation plays a major role in many physiological and pathophysiological processes. The mechanisms of LDL peroxidation induced by transition metal ions have therefore been studied intensively. It has been proposed that the mechanism involves free radical production that occurs via decomposition of hydroperoxides. This, in turn, requires the cleavage of O-H bonds. Cleavage of O-D bond is slower and comparison of the kinetics in D2O to the kinetics in H2O is therefore a straightforward way to test this aspect of the alleged mechanism. The kinetics of peroxidation exhibit marked deuterium kinetic isotope effects at all the stages of oxidation under all the studied conditions. We found that the rate of propagation of copper-induced peroxidation is a monotonically decreasing function of D2O fraction in D2O/H2O mixtures. The only elementary reaction that involves “exchangeable” hydrogen at this stage is copper-induced decomposition of conjugated hydroperoxides. Therefore, we conclude that the latter step is rate-limiting reaction including cleavage of oxygen-hydrogen bond of hydroperoxide. The lag preceding rapid peroxidation exhibits a biphasic dependence on the fraction of D2O. This may be understood on the basis of the effect of substituting hydrogen atoms by deuterium. Specifically, such substitution is expected to decrease both the rate of initiation of peroxidation and the potency of the antioxidant. We interpret our results in terms of the effects of isotopic substitution on the rates of the reactions that involve the abstraction of “exchangeable” hydrogen atoms of OH groups in tocopherol and hydroperoxides.