Oxidative modification and nitration of human low-density lipoproteins by the reaction of hypochlorous acid with nitrite.
Hypochlorous acid (HOCl) reacts with nitrite (NO2-) at a molar ratio of 1:1 yielding an equimolar amount of nitrate. The rate of this reaction follows the dissociation of hypochlorous acid and decreases with the increasing of pH from 4 to 10 as assayed by stopped-flow analysis, suggesting that HOCl, not hypochlorite, is the reactant. The second-order rate constant at pH 7.2, 25 degrees C, was estimated as (7.4 +/- 1.3) x 10(3) M(-1) s(-1), a rate considerably higher than that of the Fenton reaction (42 M(-1) s(-1)). In human low-density lipoproteins (LDL) the reaction led to a loss of beta-carotene and alpha-tocopherol. The NO2-/HOCl mixture initiated lipid peroxidation in LDL, whereas NO2- or HOCl alone had only little effect. When LDL was added immediately after mixing of NO2- with HOCl, no loss of antioxidants or accumulation of lipid peroxidation products was observed, suggesting that a short-lived reactive intermediate, previously postulated as nitryl chloride, is the reactive species. The mixture NO2-/HOCl as well as peroxynitrite led to the formation of 3-nitrotyrosine in LDL as assayed using a monoclonal anti-nitrotyrosine antibody. Furthermore, incubation of J774.2 macrophage-like cells with LDL, pretreated with the NO2-/HOCl mixture, led to increased cellular accumulation of cholesterol. Thus modification of LDL caused by the reaction of nitrite with HOCl contributes to the formation of cholesterol-rich cells, a key feature of the early atherosclerotic lesion.
Panasenko
O M
OM
Institut für Physiologische Chemie I, Heinrich-Heine-Universität Düsseldorf, Germany.
Briviba
K
K
Klotz
L O
LO
Sies
H
H
D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D006801:Humans; D006997:Hypochlorous Acid; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood / Q000737:chemistry* / Q000494:pharmacology; D008264:Macrophages; D009573:Nitrites; D010084:Oxidation-Reduction; D014810:Vitamin E / Q000032:analysis; D019207:beta Carotene / Q000032:analysis
D008077:Lipoproteins, LDL; D009573:Nitrites; C099252:oxidized low density lipoprotein; D019207:beta Carotene; D014810:Vitamin E; D006997:Hypochlorous Acid; D002784:Cholesterol