Electrophoretic mobility changes of oxidized human low density lipoprotein measured by laser Doppler electrophoresis

Laser Doppler Electrophoresis was used to detect changes in the surface charge of low density lipoprotein populations exposed to oxidative stress. Before oxidative stress, low density lipoprotein suspensions exhibited homogenous populations of net negative charge but after exposure to hydroxyl and superoxide radicals, peroxyl radicals, or by Cu2+ generated oxidants they exhibited Laser Doppler Electrophoresis changes. The major population of low density lipoprotein became more negatively charged, in agreement with agarose gel electrophoresis. However, Laser Doppler Electrophoresis detected greater heterogeneity of low density lipoprotein, compared to agarose gel electrophoresis. Partially oxidized low density lipoprotein exhibited a less negatively charged subpopulation of particles compared to control samples. This has not been reported previously. Hence, Laser Doppler Electrophoresis is a sensitive method for detecting the appearance of subpopulations of differing surface charge density in oxidatively modified low density lipoprotein. beta-carotene protected low density lipoprotein against oxidative modification even when endogenous vitamin E levels are low. Vitamin E-deficient low density lipoprotein pretreated with beta-carotene exhibited a more narrow negative population when oxidized with peroxyl radicals, compared to control. Native low density lipoprotein pretreated with a mixture of all-trans- and cis-beta-carotene was also protected.