Fluorescent Benzothiazinone Analogues Efficiently and Selectively Label Dpre1 in Mycobacteria and Actinobacteria

Raphael Sommer, João Neres, Jérémie Piton, Neeraj Dhar, Astrid van der Sar, Raju Mukherjee, Thierry Laroche, Paul J Dyson, John D McKinney, Wilbert Bitter, Vadim Makarov, Stewart T Cole

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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 languageEnglish
Pages (from-to)3184-3192
Number of pages9
JournalACS chemical biology
Issue number11
Early online date5 Oct 2018
Publication statusPublished - 16 Nov 2018


We thank A. Vocat and S. Boy-Röttger for technical assistance, L. Vera-Cabrera for providing strains, C. Trefzer and K. Johnsson for useful discussions and L. Reymond for the synthesis of compounds. The research leading to these results received funding from the European Community’s Seventh Framework Programme (MM4TB, Grant 260872) and from an International Incoming Marie Curie fellowship from the European Commission to J.N. (DPRETB, Grant 252802). R.S. was supported by a grant from the Fondation Jacqueline Beytout. V.M. and S.T.C. are named inventors on patents related to this work.

FundersFunder number
Fondation Jacqueline Beytout
International Incoming Marie Curie
Seventh Framework ProgrammeMM4TB, 260872
European Commission252802


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