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Detection of lipid radicals by electron paramagnetic resonance spin trapping using intact cells enriched with polyunsaturated fatty acid
Journal article   Open access   Peer reviewed

Detection of lipid radicals by electron paramagnetic resonance spin trapping using intact cells enriched with polyunsaturated fatty acid

James A North, Arthur A Spector and Garry R Buettner
The Journal of biological chemistry, Vol.267(9), pp.5743-5746
1992
DOI: 10.1016/S0021-9258(18)42614-2
PMID: 1313417
url
https://doi.org/10.1016/S0021-9258(18)42614-2View
Published (Version of record) Open Access

Abstract

Electron paramagnetic resonance (EPR) spin trapping was used to detect lipid-derived free radicals generated by iron-induced oxidative stress in intact cells. Using the spin trap alpha-(4-pyridyl 1-oxide)-N-tert-butylnitrone (POBN), carbon-centered radical adducts were detected. These lipid-derived free radicals were formed during incubation of ferrous iron with U937 cells that were enriched with docosahexaenoic acid (22:6n-3). The EPR spectra exhibited apparent hyperfine splittings characteristic of a POBN/alkyl radical, aN = 15.63 +/- 0.06 G and aH = 2.66 +/- 0.03 G, generated as a result of beta-scission of alkoxyl radicals. Spin adduct formation depended on the FeSO4 content of the incubation medium and the number of 22:6-enriched cells present; when the cells were enriched with oleic acid (18:1n-9), spin adducts were not detected. This is the first direct demonstration, using EPR, of a lipid-derived radical formed in intact cells in response to oxidant stress.
 Cell Physiology Fundamental and applied biological sciences. Psychology Biological and medical sciences Molecular and cellular biology Cell metabolism, cell oxidation

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