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Cited by 212

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Oxidation of Guanine in DNA by Ru(phen)2(dppz)3+ Using the Flash-Quench Technique

Stemp, E. D. A., Arkin, M. R., Barton, J. K.·1997·Journal of the American Chemical Society

Abstract

A flash-quench method has been developed to probe oxidative damage to DNA. A photoexcited Ru(II) intercalator is quenched in DNA by a weakly bound, electron-transfer quencher to generate Ru(III), a powerful ground-state oxidant. Once generated, Δ-Ru(phen) 2 (dppz) 3+ bound to poly(dG-dC) rapidly oxidizes guanine within the DNA duplex. Transient absorption spectroscopy indicates rapid formation of the neutral guanine radical within the DNA duplex. Permanent damage resulting from the flash-quench experiment is monitored by gel electrophoresis of synthetic oligonucleotide duplexes. Oxidative damage, visualized by treatment with piperidine, occurs selectively at the 5‘-G of 5‘-GG-3‘ sites and at the 5‘- and central G of 5‘-GGG-3‘ triplets; enzymatic digestion in the absence of piperidine treatment shows formation of 8-oxo-2‘-deoxyguanosine with Ru(NH 3 ) 6 3+ as quencher. The yield of base damage is, furthermore, modulated by the choice of electron-transfer quencher. Quantum yields for damage vary in the order Ru(NH 3 ) 6 3+ < methyl viologen 2+ < Co(NH 3 ) 5 Cl 2+ and correlate with the instability of the reduced quencher. The flash-quench method, combining spectroscopy and product analysis, offers a novel and tunable approach to explore electron transfer chemistry on double helical DNA.

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Metadata

Authors
Stemp, E. D. A., Arkin, M. R., & Barton, J. K.
Venue
Journal of the American Chemical Society
Published
1997-03-01
Volume
119
Issue
12
Pages
2921-2925
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
OpenAlex
W2026084209
Cited by
212
References
2
FWCI
5.06
Type
journal article
Language
en
Open access
closed

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