Interaction of metronidazole with Escherichia coli deoxyribonucleic acid

Abstract

To define the characteristics of the reported binding of metronidazole to DNA, we isolated the DNA from hypoxic incubation mixtures that contained both [14C]metronidazole and metronidazole-susceptible strains of Escherichia coli. Thus, either [2-14C]metronidazole or [1',2'-14C]metronidazole was incubated with either wild-type E. coli (strain AB1157) or a DNA repair mutant (strain SR58) that is highly susceptible to metronidazole. Approximately 0.02% of the radiolabel in the metronidazole was found to be associated with DNA isolated from both strains of bacteria, a percentage similar to that found to be associated with DNA from mammalian sources in a variety of in vitro and in vivo experiments performed by other investigators. The bound radioactivity was not diminished, however, when a great excess of non-radiolabeled metronidazole was included in the incubation mixture, indicating that the binding we observed was probably due to impurities in the radiolabeled metronidazole. We also examined the binding to DNA of a possible surrogate for the partially reduced form of metronidazole, 1-methyl-4-phenyl-5-nitrosoimidazole (5NO), that has been described previously. The binding of the tritiated form of 5NO to DNA was also found to be undiminished by the addition of carrier 5NO (a finding which does not refute the hypothesis that 5NO may serve as a surrogate for the study of the active form of metronidazole). These studies do not exclude the binding to DNA of either metronidazole or a possible surrogate of its active functionality, but they indicate that if such binding occurs, it must be limited to very few sites on DNA and hence will be difficult to characterize.