The specificity of lipoxygenase-catalyzed lipid peroxidation and the effects of radical-scavenging antioxidants.
The oxidation of low density lipoprotein (LDL) by lipoxygenase has been implicated in the pathogenesis of atherosclerosis. It has been known that lipoxygenase-mediated lipid peroxidation proceeds in general via regio-, stereo- and enantio-specific mechanisms, but that it is sometimes accompanied by a share of random hydroperoxides as side reaction products. In this study we investigated the oxidation of various substrates (linoleic acid, methyl linoleate, phosphatidylcholine, isolated LDL, and human plasma) by the arachidonate 15-lipoxygenases from rabbit reticulocytes and soybeans aiming at elucidating the effects of substrate, lipoxygenase and reaction milieu on the contribution and mechanism of random oxidation and also the effect of antioxidant. The specific character of the rabbit 15-lipoxygenase reaction was confirmed under all conditions employed here. However, the specificity by soybean lipoxygenase was markedly dependent on the conditions. When phosphatidylcholine liposomes and LDL were oxygenated by soybean lipoxygenase, the product pattern was found to be exclusively regio-, stereo-, and enantio-random. When free linoleic acid was incorporated into PC liposomes and oxidized by soybean lipoxygenase, the free acid was specifically oxygenated, whereas esterified linoleate gave random oxidation products exclusively. Radical-scavenging antioxidants such as alpha-tocopherol, ascorbic acid and 2-carboxy-2,5,7,8-tetramethyl-6-chromanol selectively inhibited the random oxidation but did not influence specific product formation. It is assumed that the random reaction products originate from free radical intermediates, which have escaped the active site of the enzyme and thus may be accessible to radical scavengers. These data indicate that the specificity of lipoxygenase-catalyzed lipid oxidation and the inhibitory effects of antioxidants depend on the physico-chemical state of the substrate and type of lipoxygenase and that they may change completely depending on the conditions.
Noguchi
Noriko
N
Research Center for Advanced Science and Technology, The University of Tokyo, Meguro, Japan.
Yamashita
Hiromasa
H
Hamahara
Jun
J
Nakamura
Akio
A
Kühn
Hartmut
H
Niki
Etsuo
E
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001093:Arachidonate 15-Lipoxygenase / Q000097:blood / Q000378:metabolism*; D002384:Catalysis; D016166:Free Radical Scavengers / Q000494:pharmacology*; D015227:Lipid Peroxidation; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology; D013025:Glycine max / Q000201:enzymology; D013379:Substrate Specificity
D000975:Antioxidants; D016166:Free Radical Scavengers; D001093:Arachidonate 15-Lipoxygenase