ESR identification of free radicals formed from the oxidation of catechol estrogens by Cu2+

Andrew M. Seacat, Periannan Kuppusamy, Jay L. Zweier, James D. Yager

Research output: Contribution to journalArticle

Abstract

Catechol estrogens are genotoxic, indirectly through redox cycling mechanisms leading to oxidative DNA damage and directly by formation of quinone-DNA adducts. Previously, we demonstrated that Cu2+ can oxidize estradiol (E2) catechols, establishing a copper redox cycle leading to the formation of DNA strand breaks. The goal of this study was to use electron spin resonance techniques to identify the free radical intermediates formed. The 2- and 4-OH catechols of E2 and ethinyl estradiol (EE) were oxidized to semiquinone intermediates, stabilized by Mg2+, when incubated with Cu2+. The 4-OH-EE semiquinone decayed more slowly than the 2-OH-EE semiquinone. Using the spin trap α-(4-pyridyl-1-oxide)-N-tert-butylnitrone, 4-OH-E2 plus Cu2+ generated hydroxyl radicals at a greater rate than 2-OH-E2 plus Cu2+. Formation of hydroxyl and methyl radical adducts was detected, using 5,5-dimethyl-1-pyrroline-N-oxide as the spin trap, when 2-OH-E2 was incubated with Cu2+ and 1% dimethyl sulfoxide. This was inhibited by the Cu2+ chelator bathocuproinedisulfonic acid and catalase. These data demonstrate that the oxidation of estrogen catechols by Cu2+ leads to a Cu-dependent mechanism of hydroxyl radical production via a hydrogen peroxide intermediate and suggest a mechanism for estrogen-associated site-specific DNA damage and mutagenesis.

Original languageEnglish (US)
Pages (from-to)45-52
Number of pages8
JournalArchives of Biochemistry and Biophysics
Volume347
Issue number1
DOIs
StatePublished - Nov 1 1997

Keywords

  • Catechol estrogens
  • Copper
  • DNA
  • Free radicals
  • Reactive oxygen
  • Redox cycle

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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