Evaluation of spin trapping agents and trapping conditions for detection of cell-generated reactive oxygen species. Arch Biochem Biophys 2005 May 01;437(1):59-68
Date
04/12/2005Pubmed ID
15820217DOI
10.1016/j.abb.2005.02.028Scopus ID
2-s2.0-16244368485 (requires institutional sign-in at Scopus site) 95 CitationsAbstract
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.
Author List
Shi H, Timmins G, Monske M, Burdick A, Kalyanaraman B, Liu Y, Clément JL, Burchiel S, Liu KJAuthor
Balaraman Kalyanaraman PhD Professor in the Biophysics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Cell LineCyclic N-Oxides
Electron Spin Resonance Spectroscopy
Humans
Reactive Oxygen Species
Sensitivity and Specificity
Spin Trapping
Superoxides
