EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives
Superoxide radical anion (O<sub>2</sub><sup>•−</sup>) and its derivatives regulate numerous physiological and pathological processes, which are extensively studied. The aim of our work was to utilize KO<sub>2</sub> as a source of O<sub>2</sub><sup&g...
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MDPI AG
2021-08-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/10/8/1286 |
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| author | Anton Misak Vlasta Brezova Miroslav Chovanec Karol Luspai Muhammad Jawad Nasim Marian Grman Lenka Tomasova Claus Jacob Karol Ondrias |
| author_facet | Anton Misak Vlasta Brezova Miroslav Chovanec Karol Luspai Muhammad Jawad Nasim Marian Grman Lenka Tomasova Claus Jacob Karol Ondrias |
| author_sort | Anton Misak |
| collection | DOAJ |
| description | Superoxide radical anion (O<sub>2</sub><sup>•−</sup>) and its derivatives regulate numerous physiological and pathological processes, which are extensively studied. The aim of our work was to utilize KO<sub>2</sub> as a source of O<sub>2</sub><sup>•−</sup> and the electron paramagnetic resonance (EPR) spin trapping 5-<i>tert</i>-butoxycarbonyl-5-methyl-1-pyrroline <i>N</i>-oxide (BMPO) technique for the preparation of <sup>•</sup>BMPO-OOH and/or <sup>•</sup>BMPO-OH radicals in water solution without DMSO. The method distinguishes the interactions of various compounds with <sup>•</sup>BMPO-OOH and/or <sup>•</sup>BMPO-OH radicals over time. Here, we show that the addition of a buffered BMPO-HCl mixture to powdered KO<sub>2</sub> formed relatively stable <sup>•</sup>BMPO-OOH and <sup>•</sup>BMPO-OH radicals and H<sub>2</sub>O<sub>2</sub>, where the <sup>•</sup>BMPO-OOH/OH ratio depended on the pH. At a final pH of ~6.5–8.0, the concentration of <sup>•</sup>BMPO-OOH radicals was ≥20 times higher than that of <sup>•</sup>BMPO-OH, whereas at pH 9.0–10.0, the <sup>•</sup>BMPO-OH radicals prevailed. The <sup>•</sup>BMPO-OOH/OH radicals effectively cleaved the plasmid DNA. H<sub>2</sub>S decreased the concentration of <sup>•</sup>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 <sup>•</sup>BMPO-OH over the <sup>•</sup>BMPO-OOH radicals. In conclusion, the presented approach of using KO<sub>2</sub> as a source of O<sub>2</sub><sup>•−</sup>/H<sub>2</sub>O<sub>2</sub> and EPR spin trap BMPO for the preparation of <sup>•</sup>BMPO-OOH/OH radicals in a physiological solution could be useful to study the biological effects of radicals and their interactions with compounds. |
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| language | English |
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| spelling | doaj.art-abe424a591c0460c9759feb3dd404ab12023-11-22T06:36:55ZengMDPI AGAntioxidants2076-39212021-08-01108128610.3390/antiox10081286EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-DerivativesAnton Misak0Vlasta Brezova1Miroslav Chovanec2Karol Luspai3Muhammad Jawad Nasim4Marian Grman5Lenka Tomasova6Claus Jacob7Karol Ondrias8Biomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, SlovakiaInstitute of Physical Chemistry and Chemical Physics, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Radlinského 9, 81237 Bratislava, SlovakiaBiomedical Research Center, Department of Genetics, Cancer Research Institute, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, SlovakiaInstitute of Physical Chemistry and Chemical Physics, Faculty of Chemical and Food Technology, Slovak University of Technology in Bratislava, Radlinského 9, 81237 Bratislava, SlovakiaDivision of Bioorganic Chemistry, School of Pharmacy, University of Saarland, D-66123 Saarbruecken, GermanyBiomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, SlovakiaBiomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, SlovakiaDivision of Bioorganic Chemistry, School of Pharmacy, University of Saarland, D-66123 Saarbruecken, GermanyBiomedical Research Center, Department of Molecular Physiology, Institute of Clinical and Translational Research, Slovak Academy of Sciences, Dúbravská Cesta 9, 84505 Bratislava, SlovakiaSuperoxide radical anion (O<sub>2</sub><sup>•−</sup>) and its derivatives regulate numerous physiological and pathological processes, which are extensively studied. The aim of our work was to utilize KO<sub>2</sub> as a source of O<sub>2</sub><sup>•−</sup> and the electron paramagnetic resonance (EPR) spin trapping 5-<i>tert</i>-butoxycarbonyl-5-methyl-1-pyrroline <i>N</i>-oxide (BMPO) technique for the preparation of <sup>•</sup>BMPO-OOH and/or <sup>•</sup>BMPO-OH radicals in water solution without DMSO. The method distinguishes the interactions of various compounds with <sup>•</sup>BMPO-OOH and/or <sup>•</sup>BMPO-OH radicals over time. Here, we show that the addition of a buffered BMPO-HCl mixture to powdered KO<sub>2</sub> formed relatively stable <sup>•</sup>BMPO-OOH and <sup>•</sup>BMPO-OH radicals and H<sub>2</sub>O<sub>2</sub>, where the <sup>•</sup>BMPO-OOH/OH ratio depended on the pH. At a final pH of ~6.5–8.0, the concentration of <sup>•</sup>BMPO-OOH radicals was ≥20 times higher than that of <sup>•</sup>BMPO-OH, whereas at pH 9.0–10.0, the <sup>•</sup>BMPO-OH radicals prevailed. The <sup>•</sup>BMPO-OOH/OH radicals effectively cleaved the plasmid DNA. H<sub>2</sub>S decreased the concentration of <sup>•</sup>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 <sup>•</sup>BMPO-OH over the <sup>•</sup>BMPO-OOH radicals. In conclusion, the presented approach of using KO<sub>2</sub> as a source of O<sub>2</sub><sup>•−</sup>/H<sub>2</sub>O<sub>2</sub> and EPR spin trap BMPO for the preparation of <sup>•</sup>BMPO-OOH/OH radicals in a physiological solution could be useful to study the biological effects of radicals and their interactions with compounds.https://www.mdpi.com/2076-3921/10/8/1286KO<sub>2</sub>antioxidantsEPR spectra simulation<sup>•</sup>BMPO-OOH spin adductsuperoxideradical |
| spellingShingle | Anton Misak Vlasta Brezova Miroslav Chovanec Karol Luspai Muhammad Jawad Nasim Marian Grman Lenka Tomasova Claus Jacob Karol Ondrias EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives Antioxidants KO<sub>2</sub> antioxidants EPR spectra simulation <sup>•</sup>BMPO-OOH spin adduct superoxide radical |
| title | EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives |
| title_full | EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives |
| title_fullStr | EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives |
| title_full_unstemmed | EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives |
| title_short | EPR Study of KO<sub>2</sub> as a Source of Superoxide and <sup>•</sup>BMPO-OH/OOH Radical That Cleaves Plasmid DNA and Detects Radical Interaction with H<sub>2</sub>S and Se-Derivatives |
| title_sort | epr study of ko sub 2 sub as a source of superoxide and sup • sup bmpo oh ooh radical that cleaves plasmid dna and detects radical interaction with h sub 2 sub s and se derivatives |
| topic | KO<sub>2</sub> antioxidants EPR spectra simulation <sup>•</sup>BMPO-OOH spin adduct superoxide radical |
| url | https://www.mdpi.com/2076-3921/10/8/1286 |
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