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

23 Citations (Scopus)

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

Fingerprint

Spin Trapping
Ethylene Glycol
Superoxides
Polyethylene glycols
Superoxide Dismutase
Reactive Oxygen Species
Neutrophils
Hydroxyl Radical
Free Radicals
Phagosomes
Vacuoles
Phagocytosis
Oxides
Paramagnetic resonance
Cytoplasm

Keywords

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

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

Cite this

Intracellular spin-trapping of oxygen-centered radicals generated by human neutrophils. / Pou, Sovitj; Rosen, Gerald M.; Britigan, Bradley E.; Cohen, Myron S.

In: BBA - General Subjects, Vol. 991, No. 3, 27.06.1989, p. 459-464.

Research output: Contribution to journalArticle

Pou, Sovitj ; Rosen, Gerald M. ; Britigan, Bradley E. ; Cohen, Myron S. / Intracellular spin-trapping of oxygen-centered radicals generated by human neutrophils. In: BBA - General Subjects. 1989 ; Vol. 991, No. 3. pp. 459-464.
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