Abstract
Benzothiazinones (BTZ) are highly potent bactericidal inhibitors of mycobacteria and the lead compound, BTZ043, and the optimized drug candidate, PBTZ169, have potential for the treatment of tuberculosis. Here, we exploited the tractability of the BTZ scaffold by attaching a range of fluorophores to the 2-substituent of the BTZ ring via short linkers. We show by means of fluorescence imaging that the most advanced derivative, JN108, is capable of efficiently labeling its target, the essential flavoenzyme DprE1, both in cell-free extracts and after purification as well as in growing cells of different actinobacterial species. DprE1 displays a polar localization in Mycobacterium tuberculosis, M. marinum, M. smegmatis, and Nocardia farcinica but not in Corynebacterium glutamicum. Finally, mutation of the cysteine residue in DprE1 in these species, to which BTZ covalently binds, abolishes completely the interaction with JN108, thereby highlighting the specificity of this fluorescent probe.
Original language | English |
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Pages (from-to) | 3184-3192 |
Number of pages | 9 |
Journal | ACS Chemical Biology |
Volume | 13 |
Issue number | 11 |
Early online date | 5 Oct 2018 |
DOIs | |
Publication status | Published - 16 Nov 2018 |
Keywords
- Actinomycetales/drug effects
- Affinity Labels/chemical synthesis
- Alcohol Oxidoreductases/antagonists & inhibitors
- Antitubercular Agents/chemical synthesis
- Bacterial Proteins/antagonists & inhibitors
- Cell Membrane/metabolism
- Drug Design
- Enzyme Inhibitors/chemical synthesis
- Fluoresceins/chemical synthesis
- Fluorescence
- Fluorescent Dyes/chemical synthesis
- Hep G2 Cells
- Humans
- Microbial Sensitivity Tests
- Microscopy, Fluorescence/methods
- Mutation
- Thiazines/chemical synthesis