DNA damage by tert-butoxyl radicals generated in the photolysis of a water-soluble, DNA-binding peroxyester acting as a radical source.
The photolysis of the water-soluble perester 1 leads to tert-butoxyl radicals as confirmed by EPR studies with the spin trap 5, 5-dimethylpyrroline N-oxide (DMPO). In the presence of DNA, oxidative cleavage of the latter was demonstrated by the formation of strand breaks in supercoiled pBR 322 DNA and by a substantial decrease of the melting temperature of salmon testes DNA. Guanidine, released from, for example, oxazolone and oxoimidazolidine on base treatment, was observed with calf thymus DNA and 2'-deoxyguanosine. These DNA modifications were effectively inhibited by the radical scavenger di-tert-butylcresol or the hydrogen atom donor glutathione. Photosensitization by the arene chromophore was excluded since the corresponding ester 2 caused no DNA damage, nor were the photoproducts of the perester 1 active. The efficacy of the perester 1 in oxidizing DNA derives from the fact that the tert-butoxyl radicals are photolytically generated in the immediate vicinity of the DNA, due to electrostatic binding of the cationic perester to the DNA, as confirmed by fluorescence measurements. These results demonstrate that the photolysis of perester 1 provides a suitable source of tert-butoxyl radicals in aqueous media, a necessary prerequisite for biochemical investigations.
Institute of Organic Chemistry, University of Würzburg, Am Hubland, D-97074 Würzburg, Germany. adam@chemie.uniwuerzburg.de
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Adam
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Grimm
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Saha-Möller
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Dall'Acqua
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Miolo
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Vedaldi
D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D004247:DNA / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism*; D004249:DNA Damage; D004278:DNA, Superhelical / Q000737:chemistry / Q000378:metabolism; D005609:Free Radicals / Q000737:chemistry; D006147:Guanine / Q000737:chemistry / Q000378:metabolism; D010084:Oxidation-Reduction; D010782:Photolysis; D000644:Quaternary Ammonium Compounds / Q000138:chemical synthesis / Q000737:chemistry* / Q000633:toxicity*
C115185:(4-(tert-butyldioxycarbonyl)benzyl)triethylammonium chloride; D004278:DNA, Superhelical; D005609:Free Radicals; D000644:Quaternary Ammonium Compounds; D006147:Guanine; D004247:DNA