Hemoglobin induced apolipoprotein B crosslinking in low-density lipoprotein peroxidation.
Oxidative modification of human low-density lipoprotein (LDL) is thought to play a major role in the development of atherosclerosis. Free hemin, hemoglobin, myoglobin, and horseradish peroxidase (HRP) were reported in different studies as promoters of LDL lipid oxidation. Based on our previous finding that hemin induced oxidative crosslinking of the LDL protein, apolipoprotein B (apo B) (Y. I. Miller and N. Shaklai (1994) Biochem. Mol. Biol. Int. 34, 1121-1129), we compared the ability of free hemin and the above hemoproteins to induce peroxidation modification of apo B using SDS-PAGE. The levels of the final products of lipid peroxidation were determined as thiobarbituric acid-reactive substances. Hemoglobin and myoglobin were found to be as active as free hemin and all these were much more active than the classic peroxidase HRP. Moreover, the products of oxidized apo B differed: hemoglobin, myoglobin, and hemin induced mostly covalent aggregates, while HRP caused fragmentation of apo B. Hemoglobin reactivity was expressed at low H2O2 concentrations even in the absence of molecular oxygen. Desferal, along with other antioxidants, inhibited the hemoglobin-induced LDL oxidation independently of its iron-chelating property. The high peroxidative reactivity of hemoglobin is explained by its ability (unlike HRP) to transfer the oxidative equivalents from the heme active site, through the globin, to LDL. The apo B radicals thus formed are terminated, yielding intermolecular crosslinked protein. It is suggested that small amounts of the highly reactive hemoglobin in plasma, suffice to trigger LDL protein oxidation (along with its lipid oxidation), thereby inflict the atherosclerosis precondition.
Miller
Y I
YI
Sackler Institute of Molecular Medicine, Sackler Faculty of Medicine, Tel-Aviv University, Ramat-Aviv, Israel.
Felikman
Y
Y
Shaklai
N
N
D000975:Antioxidants / Q000494:pharmacology; D001055:Apolipoproteins B / Q000737:chemistry / Q000378:metabolism*; D001161:Arteriosclerosis / Q000209:etiology; D003432:Cross-Linking Reagents; D006420:Hemeproteins / Q000378:metabolism; D006427:Hemin / Q000378:metabolism; D006454:Hemoglobins / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D066298:In Vitro Techniques; D008077:Lipoproteins, LDL / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction
D000975:Antioxidants; D001055:Apolipoproteins B; D003432:Cross-Linking Reagents; D006420:Hemeproteins; D006454:Hemoglobins; D008077:Lipoproteins, LDL; D006427:Hemin; D006861:Hydrogen Peroxide; D006735:Horseradish Peroxidase