Spin trapping evidence for the lack of significant hydroxyl radical production during the respiration burst of human phagocytes using a spin adduct resistant to superoxide-mediated destruction

Bradley E Britigan; Thomas J Coffman; Garry R Buettner

doi:10.1016/S0021-9258(19)39850-3

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Spin trapping evidence for the lack of significant hydroxyl radical production during the respiration burst of human phagocytes using a spin adduct resistant to superoxide-mediated destruction

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Spin trapping evidence for the lack of significant hydroxyl radical production during the respiration burst of human phagocytes using a spin adduct resistant to superoxide-mediated destruction

Bradley E Britigan, Thomas J Coffman and Garry R Buettner

The Journal of biological chemistry, Vol.265(5), pp.2650-2656

1990

DOI: 10.1016/S0021-9258(19)39850-3

PMID: 2154454

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Abstract

Failure to detect hydroxyl radical (.OH)-derived spin adducts of 5,5-dimethyl-1-pyrroline N-oxide in electron spin resonance (ESR) spin trapping experiments has been offered as evidence for the lack of the endogenous capacity of stimulated human phagocytes (neutrophils, monocytes, and monocyte-derived macrophages (MDM] to generate .OH. Recent reports that 5,5-dimethyl-1-pyrroline N-oxide spin adducts are unstable in the presence of superoxide-generating systems such as stimulated neutrophils has raised concerns regarding the sensitivity of spin trapping techniques for assessment of phagocyte free radical formation. Consequently, we have employed a new approach that uses the spin trap N-t-butyl-alpha-phenyl-nitrone (PBN) and dimethyl sulfoxide. In the presence of dimethyl sulfoxide and PBN, the formation of .OH via three different mechanisms in air-saturated aqueous solutions all yielded a single nitroxide species whose ESR peak amplitude remained stable in the presence of superoxide (.O2-). This nitroxide, which we have assigned as PBN/.OCH3, appears to be an oxygen-centered radical derived from the spin trapping of the reaction product of O2 and methyl radical. When neutrophils, monocytes, or MDM were stimulated with phorbol 12-myristate 13-acetate or opsonized zymosan in the presence of exogenous iron, catalase-inhibitable PBN/.OCH3 was the sole nitroxide detected. In the absence of exogenous iron, no nitroxide was observed, providing evidence for the lack of the endogenous capacity of neutrophils, monocytes, and MDM to generate .OH.

Cell Physiology

Fundamental and applied biological sciences. Psychology

Biological and medical sciences

Endocytosis

Molecular and cellular biology

Details

Title: Subtitle: Spin trapping evidence for the lack of significant hydroxyl radical production during the respiration burst of human phagocytes using a spin adduct resistant to superoxide-mediated destruction
Creators: Bradley E Britigan - Veterans administration medical cent., dep. internal medicine, Iowa City IA 52242, United States
Thomas J Coffman - Veterans administration medical cent., dep. internal medicine, Iowa City IA 52242, United States
Garry R Buettner - Veterans administration medical cent., dep. internal medicine, Iowa City IA 52242, United States
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.265(5), pp.2650-2656
DOI: 10.1016/S0021-9258(19)39850-3
PMID: 2154454
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: American Society for Biochemistry and Molecular Biology; Bethesda, MD
Language: English
Date published: 1990
Academic Unit: Radiation Oncology
Record Identifier: 9984047651202771

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