Copper can promote oxidation of LDL by markedly different mechanisms.Free Radic Biol Med 1998; 24(4):607-23FR
Oxidation of LDL (0.1 microM) in PBS with copper concentrations ranging from 0.03 to 10 microM, equal to 0.3-100 Cu2+/LDL, was investigated by monitoring the formation of conjugated dienes at 234 nm. With all 8 LDL samples examined, the kinetics changed strongly at submicromolar Cu2+ concentrations. Based on time-course of the formation of conjugated dienes, cholesteryl linoleate hydroxides and hydroperoxides as well as the antioxidant consumption, two oxidation types were distinguished. Type A oxidations, observed at relatively high Cu2+ concentrations of 10-100 Cu2+/ LDL, represented the conventional kinetics of LDL oxidation with an inhibition period (= lag-time) followed by a propagation phase. In contrast, type C oxidations proceeded after a negligibly short lag time followed by a distinct propagation phase. The rate of this propagation increased rapidly to 0.5 mol diene/mol LDL and then slowed down in the presence of alpha-,gamma-tocopherols and carotenoids, which were consumed faster than tocopherols. The increase in diene absorption was due to the formation of both hydroxides and hydroperoxides suggesting a high initial decomposition of hydroperoxides. At submicromolar concentrations of about 0.1 to 0.5 microM, type C and type A oxidation can be combined resulting in 4 consecutive oxidation phases, i.e. 1st inhibition and 1st propagation (belonging to type C), followed by 2nd inhibition and 2nd propagation (belonging to type A). Increasing copper concentrations lowered the 1st propagation and shortened the 2nd inhibition periods until they melted into one apparent kinetic phase. Decreasing [Cu2+] increased the 1st propagation and 2nd inhibition but lowered the 2nd propagation phase until it completely disappeared. A threshold copper concentration, denoted as Cu(lim), can be calculated as a kinetic constant based on the Cu2+-dependence for the rate of 2nd propagation. Below Cu(lim), LDL oxidation proceeds only via type C kinetics. The Cu2+-dependence of the oxidation kinetics suggests that LDL contains two different Cu2+ biding sites. Cu2+ at the low-affinity binding sites, with half-saturation at 5-50 Cu2+/LDL, initiates and accelerates the 2nd propagation by decomposing lipid hydroperoxides. Cu2+ bound to the high-affinity binding sites, with half-saturation at 0.3-2.0 Cu2+/LDL, is responsible for the 1st propagation. Arguments in favor and against this propagation being due to tocopherol mediated peroxidation (TMP) are discussed. If the lag-time concept is extended to the conjugated diene curves seen for combined oxidation profiles, then a true inhibition phase does not apply to this time interval, but instead represents the time elapsed before the onset of the 2nd propagation phase.