Hypochlorous acid (HOCl) is generated as an endogenous product of the respiratory burst in mammalian neutrophils by myeloperoxidase from hydrogen peroxide (H 2 O 2 ) and chloride (Cl Ϫ ).1) HOCl generated by myeloperoxidase is of central importance in immune surveillance and host defense mechanisms. However, it also has potential to harm normal tissue and contribute to inflammatory injury. Indeed, reagent HOCl and/or the myeloperoxidase-H 2 O 2 -Cl Ϫ system have been reported to react with nucleic acid bases to form various compounds. [2][3][4][5][6][7][8] We have examined the reaction of 2Ј-deoxyguanosine (dG) with reagent HOCl and/or the myeloperoxidase-H 2 O 2 -Cl Ϫ system and found that a diimino-imidazole nucleoside, an amino-imidazolone nucleoside, a spiroiminodihydantoin nucleoside and 8-chloro-2Ј-deoxyguanosine were generated. reported that 8-nitro-2Ј-deoxyguanosine was generated by the reaction of dG with the myeloperoxidase-H 2 O 2 -Cl Ϫ system in the presence of NO 2 Ϫ at plasma concentrations of Cl . These papers showed that NO 2 Ϫ and Br Ϫ were significant regents with respect to nucleoside damage caused by the myeloperoxidase-H 2 O 2 -Cl Ϫ system. However, in these studies and others of damage caused by myeloperoxidase, specific products and their yields were determined, but data for the whole reaction, i.e., for consumption of the target nucleosides, were extremely limited.In the present study, we investigated the reaction of a nucleoside mixture with the myeloperoxidase-H 2 O 2 -Cl Ϫ system in the presence of various anions including F , and the consumption of each nucleoside was determined by HPLC.
Results and Discussion pH-Dependence of the Reaction of the Nucleoside Mixture with the Myeloperoxidase-H 2 O 2 -Cl؊ System A nucleoside mixture (dG, dC, 2Ј-deoxythymidine (dT) and 2Ј-deoxyadenosine (dA); 100 mM each) was incubated with 50 nM myeloperoxidase in 100 mM sodium phosphate buffers of different pH with 200 mM H 2 O 2 and 100 mM NaCl at 37°C for 30 min. The reaction was terminated by addition of Nacetylcysteine. The reaction mixture was analyzed by reversed-phase HPLC to determine the remaining concentration of each nucleoside. Figure 1 shows a plot of the concentrations of nucleosides versus pH. Consumption of nucleosides was observed under mildly acidic conditions. The myeloperoxidase-H 2 O 2 -Cl Ϫ system consumed nucleosides in the following order: dGϾdCϾ ϾdTϾdAÅ0. The optimal pH for consumption of nucleosides was 4.7. At pH 4.7, total consumption was 67.3 mM corresponding to 33.7% relative to H 2 O 2 added.It has been reported that dC reacts with a myeloperoxi-