Hydrodioxyl (perhydroxyl), peroxyl, and hydroxyl radical-initiated lipid peroxidation of large unilamellar vesicles (liposomes): comparative and mechanistic studies.

The mechanisms and relative efficiencies of lipid peroxidation initiation by biological O2-derived oxidants were studied using large unilammellar vesicle (LUV) liposomes, structural models for biological membranes, as targets for oxidation. LUVs, when prepared from dilinoleoylphosphatidylcholine (DLPC) containing either 0 or 5 mol% hydroperoxide (LOOH, added either as a linoleic acid or DLPC hydroperoxide), maintained structural integrity, which enabled evaluation of the relative ability of oxidants to initiate lipid peroxidation when generated outside of the bilayer. LUVs were more oxidazable than multilamellar vesicles or lipids dispersed in solution, supporting their appropriateness as biological membrane models. In parallel to previous results using lipid dispersions (J. Aikens and T. A. Dix, 1991, J. Biol. Chem. 266, 15091-15098), both perhydroxyl (HOO.) and peroxyl (ROO.) radicals initiated lipid peroxidation in LUVs. Oxidants that did not initiate included H2O2, organic hydroperoxides, and, most notably, superoxide (O2-). HOO. and ROO. initiated by different mechanisms: HOO. required the presence of the preexisting LOOHs for efficient initiation, indicating the direct reaction of HOO. with LOOH, whereas ROO. initiated by hydrogen atom abstraction at the bisallylic site of unsaturation on the fatty acid side chain of the PCs. Hydroxyl radicals (HO.s) were poor initiators in comparison to ROO.s (and, indirectly, HOO.s), which might be considered surprising as the latter species are chemically weaker oxidants. The decreased activity of HO. was not due to decreased access to the LUVs; rather, this oxidant appears to react to generate less viable lipid peroxidation propagating species. It was also demonstrated that the fluidity of the LUV membrane had little effect on the relative initiating activity of each oxidant. It is argued that HO. may initiate lipid peroxidation only indirectly in vivo (through the generation of carbon-based peroxyl radicals, ROO.s) and that greater effort should be made to understand the roles of HOO. and ROO. at lipid peroxidation initiation.