Oxidation of methionine residues affects the structure and stability of apolipoprotein A-I in reconstituted high density lipoprotein particles.

To determine the effect of oxidative damage to lipid-bound apolipoprotein A-I (apo A-I) on its structure and stability that might be related to previously observed functional disorders of oxidized apo A-I in high density lipoproteins (HDL), we prepared homogeneous reconstituted HDL (rHDL) particles containing unoxidized apo A-I and its commonly occurring oxidized form (Met-112, 148 bis-sulfoxide). The size of the obtained discoidal rHDL particles ranged from 9.0 to 10.0 nm and did not depend upon the content of the oxidized protein. Using circular dichroism methods, no change in the secondary structure of lipid-bound oxidized apo A-I was found. Isothermal and thermal denaturation experiments showed a significant destabilization of the oxidized protein to denaturation by guanidine hydrochloride or heat. This effect was observed with and without co-reconstituted apolipoprotein A-II. Limited tryptic digestion indicated that the central region of oxidatively damaged apo A-I becomes exposed to proteolysis in the rHDL particles. Implications of these data for apolipoprotein function are discussed.