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Superoxide reaction with nitroxide spin-adducts.

A Samuni1, C M Krishna, P Riesz

  • 1Department of Molecular Biology, School of Medicine, Hebrew University of Jerusalem, Israel.

Free Radical Biology & Medicine
|January 1, 1989
PubMed
Summary
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Superoxide radicals degrade spin traps like DMPO--OH through a second-order reaction, independent of metal ions or thiols. This finding is crucial for accurately interpreting radical studies in biological systems.

Area of Science:

  • Biochemistry
  • Free Radical Chemistry
  • Spectroscopy

Background:

  • Nitroxide spin-adducts are used to detect reactive oxygen species.
  • Superoxide radical (O2-) is a key reactive oxygen species implicated in various biological processes.
  • Previous studies suggested metal ions or thiols were involved in superoxide reactions with spin traps.

Purpose of the Study:

  • To investigate the reaction kinetics and mechanism of superoxide radical with persistent nitroxide spin-adducts.
  • To clarify the role of metal ions and thiols in superoxide-mediated spin-adduct degradation.
  • To assess the implications of these reactions for interpreting electron spin resonance (ESR) studies.

Main Methods:

  • Electron spin resonance (ESR) spectrometry was used to study reactions.

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  • Superoxide radicals were generated enzymatically (xanthine/xanthine oxidase) and chemically (potassium superoxide in DMSO).
  • DMPO spin-adducts (DMPO--OH, DMPO--CH3, DMPO--H) and PBN--CH3 were reacted with superoxide.

    • Main Results:

    • Superoxide-induced depletion of DMPO--OH followed second-order kinetics.
    • The reaction did not require transition metal ions or thiols, contrary to prior reports.
    • Depleted spin-adducts could not be restored by reoxidation, indicating a non-one-electron reduction mechanism.
    • Superoxide also depleted DMPO--CH3 and DMPO--H, but not PBN--CH3.
    • Some stable nitroxides underwent pseudo-zero order depletion.

    Conclusions:

    • Superoxide radical actively destroys DMPO spin-adducts via a mechanism not involving metal ions or thiols.
    • The findings necessitate re-evaluation of ESR data in biological systems with high superoxide production.
    • Erroneous conclusions regarding primary radicals can arise from superoxide-induced spin-adduct destruction during phagocytosis or paraquat intoxication.