Nitration of gamma-tocopherol and oxidation of alpha-tocopherol by copper-zinc superoxide dismutase/H2O2/NO2-: role of nitrogen dioxide free radical

Proc Natl Acad Sci U S A. 1998 Oct 27;95(22):12912-7. doi: 10.1073/pnas.95.22.12912.

Abstract

Copper-zinc superoxide dismutase (Cu,ZnSOD) is the antioxidant enzyme that catalyzes the dismutation of superoxide (O2*-) to O2 and H2O2. In addition, Cu,ZnSOD also exhibits peroxidase activity in the presence of H2O2, leading to self-inactivation and formation of a potent enzyme-bound oxidant. We report in this study that lipid peroxidation of L-alpha-lecithin liposomes was enhanced greatly during the SOD/H2O2 reaction in the presence of nitrite anion (NO2-) with or without the metal ion chelator, diethylenetriaminepentacetic acid. The presence of NO2- also greatly enhanced alpha-tocopherol (alpha-TH) oxidation by SOD/H2O2 in saturated 1, 2-dilauroyl-sn-glycero-3-phosphatidylcholine liposomes. The major product identified by HPLC and UV-studies was alpha-tocopheryl quinone. When 1,2-diauroyl-sn-glycero-3-phosphatidylcholine liposomes containing gamma-tocopherol (gamma-TH) were incubated with SOD/H2O2/NO2-, the major product identified was 5-NO2-gamma-TH. Nitrone spin traps significantly inhibited the formation of alpha-tocopheryl quinone and 5-NO2-gamma-TH. NO2- inhibited H2O2-dependent inactivation of SOD. A proposed mechanism of this protection involves the oxidation of NO2- by an SOD-bound oxidant to the nitrogen dioxide radical (*NO2). In this study, we have shown a new mechanism of nitration catalyzed by the peroxidase activity of SOD. We conclude that NO2- is a suitable probe for investigating the peroxidase activity of familial Amyotrophic Lateral Sclerosis-linked SOD mutants.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Electron Spin Resonance Spectroscopy
  • Free Radicals / metabolism
  • Hydrogen Peroxide / metabolism*
  • Isoenzymes / metabolism
  • Isomerism
  • Kinetics
  • Liposomes
  • Nitrites / metabolism*
  • Nitrites / pharmacology
  • Nitrogen Dioxide / metabolism*
  • Oxidation-Reduction
  • Phosphatidylcholines
  • Superoxide Dismutase / metabolism*
  • Vitamin E / metabolism*

Substances

  • Free Radicals
  • Isoenzymes
  • Liposomes
  • Nitrites
  • Phosphatidylcholines
  • Vitamin E
  • Hydrogen Peroxide
  • Superoxide Dismutase
  • Nitrogen Dioxide