Inhibition of bleomycin (BLM)-induced DNA breakage by superoxide dismutase (SOD) has been reported and presumed to be due to its removal of the superoxide free radicals generated by BLM in the presence of iron(II). We have studied the possibility that the inhibitory effect might result from DNA-binding of SOD. The effect of copper-zinc SOD on BLM-induced DNA degradation was investigated using the PM-2 DNA fluorescence technique. PM-2 DNA was incubated with BLM in the presence or absence of native and heat-inactivated copperzinc SOD as determined by the epinephrine autoxidation method. The concentrations of SOD required to inhibit 50% PM-2 DNA degradation for the native and the inactivated SOD were 100 and 120 µg/ml, respectively. Analysis of the reaction mixture by agarose gel electrophoresis confirmed the absence of DNA degradation by BLM in the presence of either form of SOD. PM-2 DNA was shown to bind native or inactivated SOD by Sephadex G-100 column chromatography, fluorescence-quenching studies, and agarose gel electrophoresis. Thus, these results indicate that SOD is able to bind to PM-2 DNA and inhibit BLM-induced degradation independently of its free radical-scavenging activity. The inhibition was more effective against BLM than other compounds which degrade PM-2 DNA. This suggests that SOD may bind to BLM-binding and/or BLM degradation sites in PM-2 DNA, and the observed inhibition is unrelated to its effects on free radicals.


Research supported by a grant from Bristol Laboratories, Syracuse, N. Y.

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