EPR Study of KO2 as a Source of Superoxide and •BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H2S and Se-Derivatives
- PMID: 34439533
- PMCID: PMC8389328
- DOI: 10.3390/antiox10081286
EPR Study of KO2 as a Source of Superoxide and •BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H2S and Se-Derivatives
Abstract
Superoxide radical anion (O2•-) and its derivatives regulate numerous physiological and pathological processes, which are extensively studied. The aim of our work was to utilize KO2 as a source of O2•- and the electron paramagnetic resonance (EPR) spin trapping 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO) technique for the preparation of •BMPO-OOH and/or •BMPO-OH radicals in water solution without DMSO. The method distinguishes the interactions of various compounds with •BMPO-OOH and/or •BMPO-OH radicals over time. Here, we show that the addition of a buffered BMPO-HCl mixture to powdered KO2 formed relatively stable •BMPO-OOH and •BMPO-OH radicals and H2O2, where the •BMPO-OOH/OH ratio depended on the pH. At a final pH of ~6.5-8.0, the concentration of •BMPO-OOH radicals was ≥20 times higher than that of •BMPO-OH, whereas at pH 9.0-10.0, the •BMPO-OH radicals prevailed. The •BMPO-OOH/OH radicals effectively cleaved the plasmid DNA. H2S decreased the concentration of •BMPO-OOH/OH radicals, whereas the selenium derivatives 1-methyl-4-(3-(phenylselanyl) propyl) piperazine and 1-methyl-4-(4-(phenylselanyl) butyl) piperazine increased the proportion of •BMPO-OH over the •BMPO-OOH radicals. In conclusion, the presented approach of using KO2 as a source of O2•-/H2O2 and EPR spin trap BMPO for the preparation of •BMPO-OOH/OH radicals in a physiological solution could be useful to study the biological effects of radicals and their interactions with compounds.
Keywords: EPR spectra simulation; KO2; antioxidants; cleavage DNA; hydrogen sulfide; radical; selenium-derivatives; superoxide; •BMPO-OOH spin adduct.
Conflict of interest statement
The authors declare no conflict of interest.
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References
-
- Carroll L., Pattison D.I., Davies J.B., Anderson R.F., Lopez-Alarcon C., Davies M.J. Superoxide radicals react with peptide-derived tryptophan radicals with very high rate constants to give hydroperoxides as major products. Free Radic. Biol. Med. 2018;118:126–136. doi: 10.1016/j.freeradbiomed.2018.02.033. - DOI - PubMed
-
- Zhao H., Joseph J., Zhang H., Karoui H., Kalyanaraman B. Synthesis and biochemical applications of a solid cyclic nitrone spin trap: A relatively superior trap for detecting superoxide anions and glutathiyl radicals. Free Radic. Biol. Med. 2001;31:599–606. doi: 10.1016/S0891-5849(01)00619-0. - DOI - PubMed
-
- Bézière N., Hardy M., Poulhès F., Karoui H., Tordo P., Ouari O., Frapart Y.-M., Rockenbauer A., Boucher J.-L., Mansuy D., et al. Metabolic stability of superoxide adducts derived from newly developed cyclic nitrone spin traps. Free Radic. Biol. Med. 2014;67:150–158. doi: 10.1016/j.freeradbiomed.2013.10.812. - DOI - PubMed
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