Scavenging of Peroxynitrite by a Phenolic/peroxidase System Prevents Oxidative Damage to DNA

We examined the ability of horseradish peroxidase (HRP), an analog of human myeloperoxidase, to protect DNA against oxidative damage caused by peroxynitrite in the presence of chlorogenic acid (CGA), a naturally occurring polyphenol. Chlorogenic acid inhibits the formation of single strand breaks in supercoiled pBR322 DNA by acting as a scavenger of peroxynitrite. Horseradish peroxidase markedly enhances the extent of DNA protection by catalyzing the decomposition of peroxynitrite in the presence of CGA. Horseradish peroxidase alone does not inhibit peroxynitrite-induced DNA strand breaks, indicating that CGA is required as an electron donor to regenerate the active enzyme. The apparent second order rate constant for the HRP-mediated oxidation of CGA in the presence of peroxynitrite at pH 6.9 is 3.4 x 10(7) M(-1) s(-1). This high rate suggests that CGA and other dietary polyphenols might efficiently scavenge peroxynitrite in peroxidase-containing systems in vivo.