title: Breaking the dogma: PCB-derived semiquinone free radicals do not form covalent adducts with DNA, GSH, and amino acids.
abstract: Covalent bond formations of free radical metabolites with biomolecules like DNA and proteins are thought to constitute a major mechanism of toxicity and carcinogenesis. Glutathione (GSH) is generally accepted as a radical scavenger protecting the cell. In the present study, we investigated a semiquinone radical (SQ(●-)) metabolite of the semivolatile 4-chlorobiphenyl, using electron paramagnetic resonance spectroscopy, and oxygen consumption. Proton nuclear magnetic resonance ((1)H NMR) and liquid chromatography-mass spectrometry (LC-MS) were also employed to elucidate the radical interaction with DNA, amino acids, and GSH. We found that DNA and oligonucleotides stabilized SQ(●-) by electron delocalization in the π-stacking system, resulting in persistent radical intercalated, rather than forming a covalent bond with SQ(●-). This finding was strongly supported by the semiempirical calculation of the semioccupied molecular orbital and the linear combination of the atomic orbitals, indicating 9.8 kcal mol(-1) energy gain. The insertion of SQ(●-) into the DNA strand may result in DNA strand breaks and interruption of DNA replication process or even activate radical mediated secondary reactions. The presence of amino acids resulted in a decrease of the electron paramagnetic resonance (EPR) signal of SQ(●-) and correlated with their isoelectric points. The pH shifts the equilibrium of the dianions of hydroquinone and influenced indirectly the formation of SQ(●-). Similar findings were observed with GSH and Cys. GSH and Cys functioned as indirect radical scavengers; their activities depend on their chemical equilibria with the corresponding quinones, and their further reaction via Michael addition. The generally accepted role of GSH as radical scavenger in biological systems should be reconsidered based upon these findings, questioning the generally accepted view of radical interaction of semiquinones with biologically active compounds, like DNA, amino acids, proteins, and radical scavengers like GSH.
authors.affiliation: Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, 100 Oakdale Campus, Iowa, IA, 52242, USA.|Department of Occupational and Environmental Health, The University of Iowa, 100 Oakdale Campus, Iowa, IA, 52242, USA.|Sirindhorn College of Public Health, BanSuan, Muang, Chonburi, 2000, Thailand.
authors.forename: Orarat
authors.initials: O
authors.lastname: Wangpradit
authors.affiliation: Department of Chemistry, City College of New York, Convent Avenue at 138th St., New York, NY, 10031, USA.
authors.forename: Asif
authors.initials: A
authors.lastname: Rahaman
authors.affiliation: Central High-Field NMR Research Facility, Department of Chemistry, The University of Iowa, Iowa, IA, 52242, USA.
authors.forename: S V Santhana
authors.initials: SV
authors.lastname: Mariappan
authors.affiliation: Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, 100 Oakdale Campus, Iowa, IA, 52242, USA.|Free Radical and Radiation Biology Program, The University of Iowa, Iowa, IA, 52242, USA.
authors.forename: Garry R
authors.initials: GR
authors.lastname: Buettner
authors.affiliation: Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, 100 Oakdale Campus, Iowa, IA, 52242, USA.|Department of Occupational and Environmental Health, The University of Iowa, 100 Oakdale Campus, Iowa, IA, 52242, USA.
authors.forename: Larry W
authors.initials: LW
authors.lastname: Robertson
authors.affiliation: Interdisciplinary Graduate Program in Human Toxicology, The University of Iowa, 100 Oakdale Campus, Iowa, IA, 52242, USA. g.luthe@luthe-pharma.com.|School of Life Science, Engineering & Design, Saxion University of Applied Sciences, Enschede, Netherlands. g.luthe@luthe-pharma.com.|Luthe-Pharma, Fabrikstrasse 3, 48599, Gronau. g.luthe@luthe-pharma.com.
authors.forename: Gregor
authors.initials: G
authors.lastname: Luthe
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D016227:Benzoquinones / Q000737:chemistry*; D004247:DNA / Q000737:chemistry*; D018736:DNA Adducts / Q000737:chemistry; D004249:DNA Damage; D004578:Electron Spin Resonance Spectroscopy; D005978:Glutathione / Q000378:metabolism; D015394:Molecular Structure; D011078:Polychlorinated Biphenyls / Q000737:chemistry*; D011809:Quinones / Q000737:chemistry
chemical_list: D000596:Amino Acids; D016227:Benzoquinones; D018736:DNA Adducts; D011809:Quinones; C025232:semiquinone radicals; D004247:DNA; D011078:Polychlorinated Biphenyls; D005978:Glutathione
keywords: Carcinogenesis; DNA; Electron paramagnetic resonance; Glutathione; Polychlorinated biphenyls; Prostaglandin H synthase-2; Reactive oxygen species; Semiquinone free radicals