title: Reaction of xanthine oxidase-derived oxidants with lipid and protein of human plasma.
abstract: Xanthine oxidase and purines have recently been detected in the circulation during acute viral infection and following hepatotoxicity and shock. Reactions of xanthine oxidase-generated oxidants with human plasma or bovine serum albumin (BSA) and egg phosphatidylcholine (PC) liposomes have been studied by measuring protein sulfhydryl oxidation and two markers of free radical-mediated lipid peroxidation, thiobarbituric acid reactive substances (TBARS) and conjugated dienes. Plasma incubated with 5 mU/ml xanthine oxidase (XO) and 0.5 mM hypoxanthine (Hx) for 2 h at 37 degrees C had 25-53% oxidation of sulfhydryl groups, with greater than 80% of the oxidation occurring during the first 20 min of the reaction. Concentrations of BSA similar to those present in serum, when exposed to XO/Hx-mediated oxidative stress, showed an even greater decrease in sulfhydryl concentration than that of plasma. No significant increase in plasma TBARS and conjugated dienes was observed during the 2-h incubation period in the presence of XO. Egg PC liposomes, suspended to a plasma phospholipid-equivalent concentration, showed a minor increase in TBARS and conjugated dienes under similar XO/Hx incubation conditions. In the presence of 0.23 mM BSA, lipid peroxidation was completely inhibited. A similar inhibition of lipid peroxidation was induced by cysteine but not by uric acid. Electrophoretic and arsenite-mediated sulfur reduction analysis revealed that BSA was oxidized beyond the disulfide form, with sulfenic acid formed during the initial period of oxidation. Protein sulfhydryls served as sacrificial antioxidants, preventing plasma lipid peroxidation, as well as being targets for oxidative damage. Plasma protein thiol oxidation was determined to be a more sensitive and specific indication of oxidant stress to the vascular compartment than assessment of lipid oxidation byproducts.
authors.affiliation: Department of Anesthesiology, University of Alabama, Birmingham 35233.
authors.forename: R
authors.initials: R
authors.lastname: Radi
authors.forename: K M
authors.initials: KM
authors.lastname: Bush
authors.forename: T P
authors.initials: TP
authors.lastname: Cosgrove
authors.forename: B A
authors.initials: BA
authors.lastname: Freeman
mesh_terms: D000328:Adult; D001151:Arsenic / Q000494:pharmacology; D018053:Arsenites; D001798:Blood Proteins / Q000378:metabolism*; D006801:Humans; D019271:Hypoxanthine; D007042:Hypoxanthines / Q000494:pharmacology; D007700:Kinetics; D015227:Lipid Peroxidation; D008055:Lipids / Q000097:blood*; D008081:Liposomes; D010713:Phosphatidylcholines; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013858:Thiobarbiturates; D014969:Xanthine Oxidase / Q000097:blood* / Q000494:pharmacology
chemical_list: D018053:Arsenites; D001798:Blood Proteins; D007042:Hypoxanthines; D008055:Lipids; D008081:Liposomes; D010713:Phosphatidylcholines; D013858:Thiobarbiturates; D012710:Serum Albumin, Bovine; D019271:Hypoxanthine; D014969:Xanthine Oxidase; C029684:thiobarbituric acid; C015001:arsenite; D001151:Arsenic