Abstract
Copper is both an essential nutrient and a toxic element able to catalyze free radicals formation which damage lipids and proteins. Although the available copper redox species in aerobic environment is Cu(II), proteins that participate in metal homeostasis use Cu(I). With isolated Escherichia coli membranes, we have previously shown that electron flow through the respiratory chain promotes cupric ions reduction by NADH dehydrogenase-2 and quinones. Here, we determined Cu(II)-reductase activity by whole cells using strains deficient in these respiratory chain components. Measurements were done by the appearance of Cu(I) in the supernatants of cells exposed to sub-lethal Cu(II) concentrations. In the absence of quinones, the Cu(II)-reduction rate decreased ~70% in respect to the wild-type strain, while this diminution was about 85% in a strain lacking both NDH-2 and quinones. The decrease was ~10% in the absence of only NDH-2. In addition, we observed that quinone deficient strains failed to grow in media containing either excess or deficiency of copper, as we have described for NDH-2 deficient mutants. Thus, the Cu(II)-reduction by E. coli intact cells is mainly due to quinones and to a lesser extent to NDH-2, in a quinone-independent way. To our knowledge, this is the first in vivo demonstration of the involvement of E. coli respiratory components in the Cu(II)-reductase activity which contributes to the metal homeostasis.
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Acknowledgments
We specially thank Dr E. M. Massa, who has been the mentor of our investigations in the subject, and the lab colleagues M. R. Rintoul, L. A. Schurig-Briccio, J. M. Villegas and L. Cerioni for helpful discussions. We also thank Dr B. J. Wallace for providing strains AN387, Dr R. A. Schmitz for strain RAS50, Dr C. Rensing for strain GR6, Dr J. Imlay for providing strains SSK1, SSK2 and SSK3 and Dr I. R. Booth for providing strain MJF335. This research was supported by grants from CIUNT, CONICET, and ANPCyT (Argentine).
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Volentini, S.I., Farías, R.N., Rodríguez-Montelongo, L. et al. Cu(II)-reduction by Escherichia coli cells is dependent on respiratory chain components. Biometals 24, 827–835 (2011). https://doi.org/10.1007/s10534-011-9436-3
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DOI: https://doi.org/10.1007/s10534-011-9436-3