Trichomonas vaginalis: metronidazole and other nitroimidazole drugs are reduced by the flavin enzyme thioredoxin reductase and disrupt the cellular redox system. Implications for nitroimidazole toxicity and resistance

• Published in: Molecular microbiology Vol. 72; no. 2; pp. 518 - 536
• Main Authors: Leitsch, David, Kolarich, Daniel, Binder, Marina, Stadlmann, Johannes, Altmann, Friedrich, Duchêne, Michael
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01.04.2009; Blackwell Publishing Ltd; Blackwell
• Subjects: Animals; Antibiotics; Antiprotozoal Agents - metabolism; Antiprotozoal Agents - pharmacology; Biological and medical sciences; Cells; Cells, Cultured; Drug Resistance; Entamoeba histolytica; Enzymes; Flavins - metabolism; Fundamental and applied biological sciences. Psychology; Furazolidone - metabolism; Furazolidone - pharmacology; Giardia intestinalis; Metronidazole - metabolism; Metronidazole - pharmacology; Microbiology; Oxidation-Reduction; Parasitic protozoa; Proteomics; Protozoan Proteins - metabolism; Sulfhydryl Compounds - metabolism; Thioredoxin-Disulfide Reductase - isolation & purification; Thioredoxin-Disulfide Reductase - metabolism; Toxicity; Trichomonas vaginalis; Trichomonas vaginalis - drug effects; Trichomonas vaginalis - enzymology; Trichomonas vaginalis; Nitro compound; Imidazole derivatives; Protozoa; Microbiology; Enzyme; Toxicity; Trichomonadida; Thioredoxin; Resistance; Antiprotozoal agent; Antifungal agent; Parasiticide; Oxidoreductases; Metronidazole; Reductase
• ISSN: 0950-382X; 1365-2958
• DOI: 10.1111/j.1365-2958.2009.06675.x

Infections with the microaerophilic parasite Trichomonas vaginalis are treated with the 5-nitroimidazole drug metronidazole, which is also in use against Entamoeba histolytica, Giardia intestinalis and microaerophilic/anaerobic bacteria. Here we report that in T. vaginalis the flavin enzyme thioredoxin reductase displays nitroreductase activity with nitroimidazoles, including metronidazole, and with the nitrofuran drug furazolidone. Reactive metabolites of metronidazole and other nitroimidazoles form covalent adducts with several proteins that are known or assumed to be associated with thioredoxin-mediated redox regulation, including thioredoxin reductase itself, ribonucleotide reductase, thioredoxin peroxidase and cytosolic malate dehydrogenase. Disulphide reducing activity of thioredoxin reductase was greatly diminished in extracts of metronidazole-treated cells and intracellular non-protein thiol levels were sharply decreased. We generated a highly metronidazole-resistant cell line that displayed only minimal thioredoxin reductase activity, not due to diminished expression of the enzyme but due to the lack of its FAD cofactor. Reduction of free flavins, readily observed in metronidazole-susceptible cells, was also absent in the resistant cells. On the other hand, iron-depleted T. vaginalis cells, expressing only minimal amounts of PFOR and hydrogenosomal malate dehydrogenase, remained fully susceptible to metronidazole. Thus, taken together, our data suggest a flavin-based mechanism of metronidazole activation and thereby challenge the current model of hydrogenosomal activation of nitroimidazole drugs.