Membrane peroxidation: inhibiting effects of water-soluble antioxidants on phospholipids of different charge types

Quantitative kinetic methods of autoxidation are used to determine the antioxidant activities of two water-soluble antioxidatants of the chromanol type, 6-hydroxy-2,5,7, 8-tetramethylchroman-2-carboxylic acid (Trolox) and 6-hydroxy-2,5,7,8- tetramethyl-2-N,N,N-trimethylethanaminium methylbenzene-sulfonate (MDL 73404), during free radical peroxidation of phospholipid membranes of different charge types. The stoichiometric factor (n) for peroxyl radical trapping for both Trolox and MDL 73404 was found to be 2. Trolox was found to partition partially, approximately 20%, into the lipid phase of liposomes. The antioxidant activity of Trolox during peroxidation of membranes determined by measurements of the absolute rate constant for inhibition of oxygen uptake, kinh, was found to vary with the membrane surface charge that is controlled by variation in pH. When peroxidation is initiated in the lipid phase by azo-bis-2,4- dimethylvaleronitrile (ADVN), using a typical zwitterionic liposome, dilinoleoylphosphatidyl choline (DLPC), the kinh was found to be 2.98 X 10(3) M-1s-1. The kinh of Trolox increased approximately 2-fold for membranes that have a positive surface, including DLPC at pH 4, DLPC containing stearylamine at pH 7, and for a membrane of dimyristoylphosphatidyl acid containing linoleic acid (DMPA/LA). Conversely, Trolox does not inhibit peroxidation of negatively charged dilinoleoylphosphatidyl glycerol (DLPG) at pH 7-11. Studies made of the positively charged MDL 73404 show that its antioxidant activity using DLPC and DLPG is pH dependent. Trolox inhibits the peroxidations of DLPC initiated in the aqueous phase by azo-bis-(2-amidinopropane-HCl)(ABAP) at pH 4 or 7. However, Trolox does not inhibit the peroxidation of DLPG at pH 7. The different antioxidant activities of Trolox and MDL 73404 are rationalized in terms of a peroxyl-radical diffusion model and specific charge interactions between antioxidants and membrane surface.