Amyloid beta peptides do not form peptide-derived free radicals spontaneously, but can enhance metal-catalyzed oxidation of hydroxylamines to nitroxides

J Biol Chem. 1999 Apr 2;274(14):9392-9. doi: 10.1074/jbc.274.14.9392.

Abstract

Amyloid beta (Abeta) peptides play an important role in the pathogenesis of Alzheimer's disease. Free radical generation by Abeta peptides was suggested to be a key mechanism of their neurotoxicity. Reports that neurotoxic free radicals derived from Abeta-(1-40) and Abeta-(25-35) peptides react with the spin trap N-tert-butyl-alpha-phenylnitrone (PBN) to form a PBN/.Abeta peptide radical adduct with a specific triplet ESR signal assert that the peptide itself was the source of free radicals. We now report that three Abeta peptides, Abeta-(1-40), Abeta-(25-35), and Abeta-(40-1), do not yield radical adducts with PBN from the Oklahoma Medical Research Foundation (OMRF). In contrast to OMRF PBN, incubation of Sigma PBN in phosphate buffer without Abeta peptides produced a three-line ESR spectrum. It was shown that this nitroxide is di-tert-butylnitroxide and is formed in the Sigma PBN solution as a result of transition metal-catalyzed auto-oxidation of the respective hydroxylamine present as an impurity in the Sigma PBN. Under some conditions, incubation of PBN from Sigma with Abeta-(1-40) or Abeta-(25-35) can stimulate the formation of di-tert-butylnitroxide. It was shown that Abeta peptides enhanced oxidation of cyclic hydroxylamine 1-hydroxy-4-oxo-2,2,6, 6-tetramethylpiperidine (TEMPONE-H), which was strongly inhibited by the treatment of phosphate buffer with Chelex-100. It was shown that ferric and cupric ions are effective oxidants of TEMPONE-H. The data obtained allow us to conclude that under some conditions toxic Abeta peptides Abeta-(1-40) and Abeta-(25-35) enhance metal-catalyzed oxidation of hydroxylamine derivatives, but do not spontaneously form peptide-derived free radicals.

MeSH terms

  • Amyloid beta-Peptides / metabolism*
  • Butanes
  • Catalysis
  • Chromatography, High Pressure Liquid
  • Copper / metabolism
  • Cyclic N-Oxides
  • Ferric Compounds / metabolism
  • Free Radicals / metabolism
  • Hydroxylamines / metabolism*
  • Metals / metabolism*
  • Nitrogen Oxides / metabolism*
  • Oxidation-Reduction
  • Piperidines
  • Spin Labels

Substances

  • Amyloid beta-Peptides
  • Butanes
  • Cyclic N-Oxides
  • Ferric Compounds
  • Free Radicals
  • Hydroxylamines
  • Metals
  • Nitrogen Oxides
  • Piperidines
  • Spin Labels
  • di-t-butyl nitroxide
  • 1-hydroxy-2,2,6,6-tetramethyl-4-oxopiperidine
  • phenyl-N-tert-butylnitrone
  • Copper