Evaluation of spin trapping agents and trapping conditions for detection of cell-generated reactive oxygen species
| Autor: | Scott W. Burchiel, Jean-Louis Clément, Andrew D. Burdick, Yang Liu, Honglian Shi, Graham S. Timmins, Michael L. Monske, Ke Jian Liu, Balaraman Kalyanaraman |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
chemistry.chemical_classification
Reactive oxygen species Spin trapping Biophysics Electron Spin Resonance Spectroscopy Trapping Photochemistry Biochemistry Sensitivity and Specificity law.invention Quinone Adduct Ion Cell Line Cyclic N-Oxides chemistry.chemical_compound chemistry law Superoxides Pyrene Humans Electron paramagnetic resonance Reactive Oxygen Species Molecular Biology Spin Trapping |
| Zdroj: | Archives of biochemistry and biophysics. 437(1) |
| ISSN: | 0003-9861 |
| Popis: | Electron paramagnetic resonance with spin trapping is a useful technique to detect reactive oxygen species, such as superoxide radical anion (O2*-), a key species in many biological processes. We evaluated the abilities of four spin traps in trapping cell-generated O2*-: 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), 2-diethoxyphosphoryl-2-phenethyl-3,4-dihydro-2H-pyrrole N-oxide (DEPPEPO), 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide (DEPMPO), and 5,5-dimethyl-1-pyrroline N-oxide (DMPO). Optimal experimental conditions for obtaining maximal signal intensity of O2*- adduct in a cellular system were first studied. The maximal intensities of BMPO, DEPMPO, and DMPO adducts were similar while DEPPEPO did not trap cell-generated O2*- induced by 1,6-benzo[a]pyrene quinone in a human mammary epithelial cell line (MCF-10A). BMPO and DEPMPO adducts were more stable, considering the stability of their maximal signal, than DMPO adduct in the tested cellular systems. In addition, we observed that O2*- spin adducts were reduced to their corresponding hydroxyl adducts in the cellular system. The selection of optimal spin trap in trapping cell-generated O2*- is discussed. |
| Databáze: | OpenAIRE |
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