Intracellular spin-trapping of oxygen-centered radicals generated by human neutrophils

Sovitj Pou, Gerald M. Rosen, Bradley E. Britigan, Myron S. Cohen

Research output: Contribution to journalArticle

24 Scopus citations

Abstract

Phagocytosing neutrophils secrete superoxide into a vacuole generally inaccessible for direct study. However, the spin-trapping agent 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) enters the cytoplasm of several cell types where it can report free radical species including superoxide and hydroxyl radical. In the present study we employed a variety of experimental conditions to eliminate extracellular ESR signals and/or free radicals generated by stimulated neutrophils so that DMPO adducts reported events inside the cell. We identified a concentration of poly(ethylene glycol)-modified superoxide dismutase that permitted measurement of intracellular superoxide as determined by several criteria. It seems likely that poly(ethylene glycol)-modified superoxide dismutase is too large to enter the neutrophil phagosome. Under these conditions no hydroxyl radical was detected, as would be predicted from earlier studies with spin-trapping. Use of poly(ethylene glycol)-modified superoxide dismutase should allow on-line measurement of phogosomal events, thereby improving our understanding of microbicidal and inflammatory processes.

Original languageEnglish (US)
Pages (from-to)459-464
Number of pages6
JournalBBA - General Subjects
Volume991
Issue number3
DOIs
StatePublished - Jun 27 1989

Keywords

  • (Human neutrophils)
  • Free radical
  • Poly(ethylene glycol)
  • Spin trapping
  • Superoxide dismutase

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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