Physical Sciences and Mathematics Commons^™

Iron-Dithiothreitol Dependent Production Of 8-Hydroxy-2'-Deoxyguanosine In Dna, David Allan Cook

Undergraduate Honors Capstone Projects

Oxidative damage to DNA in a metal-thiol system was studied. Calf thymus DNA was incubated in an iron(IIl)-dithiothreitol system prepared in various buffers. Damage was measured by monitoring the production of 8-hydroxy-2'-deoxyguanosine. DNA oxidation was proportional to the concentration of dithiothreitol (DTT) in HEPES and tris(hydroxymethyl)aminomethane (Tris) buffers while it was only roughly related to DTT concentration in sodium chloride. Incubation in potassium phosphate buffer produced no damage. The results indicate that hydroxyl radicals may be generated by the iron-OTT system in HEPES, Tris, and sodium chloride, while in the phosphate buffer an iron phosphate complex may be formed which …