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Binding of Azole Antibiotics to Staphylococcus aureus Flavohemoglobin Increases Intracellular Oxidative Stress
Citation key ISI:000274891300007
Author Nobre, Ligia S. and Todorovic, Smilja and Tavares, Ana Filipa N. and Oldfield, Eric and Hildebrandt, Peter and Teixeira, Miguel and Saraiva, Ligia M.
Pages 1527-1533
Year 2010
ISSN 0021-9193
DOI 10.1128/JB.01378-09
Address 1752 N ST NW, WASHINGTON, DC 20036-2904 USA
Journal J. Bacteriology
Volume 192
Number 6
Month MAR 15
Abstract In this work, we report that flavohemoglobin contributes to the azole susceptibility of Staphylococcus aureus. We first observed that deletion of the flavohemoglobin gene leads to an increase in the viability of imidazole-treated S. aureus cells and that reversion to the wild-type phenotype occurs upon expression of flavohemoglobin from a multicopy plasmid. Further spectroscopic analyses showed that miconazole, the most efficient azole antibiotic against S. aureus, ligates to heme of both oxidized and reduced flavohemoglobin. The binding of miconazole to oxidized flavohemoglobin, with an association constant of 1.7 x 10(6) M-1, typical of a tight, specific binding equilibrium, results in augmentation of the superoxide production by the enzyme. These results are corroborated by in vivo studies showing that imidazole-treated S. aureus cells expressing flavohemoglobin contain a larger amount of reactive oxygen species. Moreover, it was observed that the survival of miconazole-treated S. aureus internalized by murine macrophages is higher for cells lacking flavohemoglobin. Altogether, the present data revealed that in S. aureus, flavohemoglobin enhances the antimicrobial activity of imidazoles via an increase of intracellular oxidative stress.
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