New Insights into the Reaction Paths of Hydroxyl Radicals with Purine Moieties in DNA and Double-Stranded Oligodeoxynucleotides

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.