Unusual copper-induced sensitization of the biological damage due to superoxide radicals.

Abstract

The role of superoxide radicals in biological damage in the presence of copper(II) ions has been investigated. Solutions of purified penicillinase in phosphate buffer, saturated with either air N/sub 2/, N/sub 2/O, or N/sub 2/O/O/sub 2/, were..gamma..-irradiated in the presence and in the absence of formate. The residual activity of the enzyme in catalyzing the cleavage of the..beta..-lactam ring of benzylpenicillin, was then monitored. The results indicate that in metal-free systems only the primary water-derived radicals, H,e/sub aq/, and OH, contribute toward enzyme inactivation, while the secondary species O/sub 2/ and CO/sub 2/ do not. The effect of copper(II) ions on the radiation-induced damage depended on which of the active species was predominant. With OH radicals, no effect of copper was detected. The damage originating from e/sub aq/ and H radicals decreased with the addition of copper, presumably due to their trapping by the copper(II) ions. In contrast, with O/sub 2/ radicals predominant in the system, copper dramatically enhanced the damage. This copper-induced sensitization was further increased in the presence of H/sub 2/O/sub 2/ and the dose modifying factor of copper exceeded 100, i.e. enchancement of the damage by two orders of magnitude, as compared with an oxygen enhancement ratiomore » of 2-3 generally found for molecular oxygen. This copper effect could be completely eliminated by EDTA. The present results suggest that the presence of both transition metal ions and H/sub 2/O/sub 2/ and the binding of the metal ions to the target bimolecules are required for the manifestation.« less