Journal
MOLECULES
Volume 24, Issue 21, Pages -Publisher
MDPI
DOI: 10.3390/molecules24213860
Keywords
DNA damage; 5 ',8-cyclopurines; 8-oxo-dG; free radicals; pulse radiolysis; gamma radiolysis; Fenton reaction; oligonucleotides
Funding
- Marie Sklodowska-Curie European Training Network (ETN) ClickGene: Click Chemistry for Future Gene Therapies to Benefit Citizens, Researchers and Industry [H2020-MSCAETN-2014-642023]
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The reaction of hydroxyl radical (HO center dot) with DNA produces many primary reactive species and many lesions as final products. In this study, we have examined the optical spectra of intermediate species derived from the reaction of HO center dot with a variety of single- and double-stranded oligodeoxynucleotides and ct-DNA in the range of 1 mu s to 1 ms by pulse radiolysis using an Intensified Charged Coupled Device (ICCD) camera. Moreover, we applied our published analytical protocol based on an LC-MS/MS system with isotopomeric internal standards to enable accurate and precise measurements of purine lesion formation. In particular, the simultaneous measurement of the four purine 5 ',8-cyclo-2 '-deoxynucleosides (cPu) and two 8-oxo-7,8-dihydro-2 '-deoxypurine (8-oxo-Pu) was obtained upon reaction of genetic material with HO center dot radicals generated either by gamma-radiolysis or Fenton-type reactions. Our results contributed to the debate in the literature regarding absolute level of lesions, method of HO center dot radical generation, 5 ' R/5 ' S diastereomeric ratio in cPu, and relative abundance between cPu and 8-oxo-Pu.
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