PMID: 9443069Jan 27, 1998Paper

The effects of different buffers on the oxidation of DNA by thiols and ferric iron

Journal of Biochemical and Molecular Toxicology
N Spear, S D Aust

Abstract

Because buffers can act as metal ligands, they can effect several reactions necessary for DNA oxidation by ferric iron and thiols, such as iron reduction. Therefore, these reactions were studied in Hepes and phosphate buffers and unbuffered NaCl. Reduction of Fe3+ by dithiothreitol (DTT) and cysteine was observed in either Hepes or NaCl solutions, but not in phosphate buffer. Thiyl radicals were observed in Hepes, but there was much less thiyl radical production in the saline or phosphate solutions. Redox cycling between either DTT or cysteine and Fe3+ also resulted in dioxygen consumption in Hepes buffer. Reduction of Fe3+ and O2 resulted in the formation of an oxidant capable of producing 8-hydroxy-2'-deoxyguanosine (8-OHdG) in calf-thymus DNA. The highest levels of 8-OHdG were detected when DTT or cysteine and Fe3+ were incubated in Hepes, while much less DNA oxidation was detected when the experiment was done in a saline solution, and almost no DNA oxidation occurred in the phosphate buffer. These results demonstrate that the use of different buffers can greatly affect the ability of thiols to promote iron-dependent oxidations.

Citations

Oct 18, 2000·Free Radical Biology & Medicine·K B BeckmanB N Ames
Mar 18, 2008·Free Radical Biology & Medicine·Marina E SolovievaVladimir S Akatov
Jun 22, 2018·Luminescence : the Journal of Biological and Chemical Luminescence·Chao WangWeiying Lin
Apr 3, 2001·Journal of Biochemical and Molecular Toxicology·Z DjuricG M Strasburg
Mar 20, 2010·Organic & Biomolecular Chemistry·Baocun ZhuWeihong Tan
Apr 28, 2021·Analytical Methods : Advancing Methods and Applications·Abdul Hadi MehmoodWeiying Lin

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