Projects per year
Abstract
The emergence of drug-resistant strains of Mycobacterium tuberculosis (Mtb) ensures that drug discovery efforts remain at the forefront of TB research. There are multiple different experimental approaches that can be employed in the discovery of anti-TB agents. Notably, inhibitors of MmpL3 are numerous and structurally diverse in Mtb and have been discovered through the generation of spontaneous resistant mutants and subsequent whole genome sequencing studies. However, this approach is not always reliable and can lead to incorrect target assignment and requires orthogonal confirmatory approaches. In fact, many of these inhibitors have also been shown to act as multi-target agents, with secondary targets in Mtb, as well as in other non-MmpL3-containing pathogens. Herein, we have investigated further the cellular targets of the MmpL3-inhibitor BM212 and a number of BM212 analogues. To determine the alternative targets of BM212, which may have been masked by MmpL3 mutations, we have applied a combination of chemo-proteomic profiling using bead-immobilised BM212 derivatives and protein extracts, along with whole-cell and biochemical assays. The study identified EthR2 (Rv0078) as a protein that binds BM212 analogues. We further demonstrated binding of BM212 to EthR2 through an in vitro tryptophan fluorescence assay, which showed significant quenching of tryptophan fluorescence upon addition of BM212. Our studies have demonstrated the value of revisiting drugs with ambiguous targets, such as MmpL3, in an attempt to find alternative targets and the study of off-target effects to understand more precisely target engagement of new hits emerging from drug screening campaigns.
Original language | English |
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Article number | 100068 |
Number of pages | 12 |
Journal | The Cell Surface |
Volume | 7 |
Early online date | 23 Nov 2021 |
DOIs | |
Publication status | Published - Dec 2021 |
Bibliographical note
Funding Information:The research leading to the results has received funding from the People Programme (Marie Curie Actions) of the European Union's Seventh Framework Programme (FP7/2007-2013) under REA grant agreement no. 291799 through the Tres Cantos Open Lab Foundation (Project Code TC206) to AC. GSB also acknowledges support in the form of a Personal Research Chair from Mr James Bardrick, a Royal Society Wolfson Research Merit Award, The Medical Research Council (MR/S000542/1, MR/R001154/1 and MR/P015859/1). We also gratefully acknowledge the inspiring scientific discussions with the GSK DDW staff as well as the support provided by the TCOLF operations team during all stages of the project.
Funding Information:
The research leading to the results has received funding from the People Programme (Marie Curie Actions) of the European Union’s Seventh Framework Programme (FP7/2007-2013) under REA grant agreement no. 291799 through the Tres Cantos Open Lab Foundation (Project Code TC206) to AC. GSB also acknowledges support in the form of a Personal Research Chair from Mr James Bardrick, a Royal Society Wolfson Research Merit Award, The Medical Research Council (MR/S000542/1, MR/R001154/1 and MR/P015859/1). We also gratefully acknowledge the inspiring scientific discussions with the GSK DDW staff as well as the support provided by the TCOLF operations team during all stages of the project.
Publisher Copyright:
© 2021 The Author(s)
Keywords
- Mycobacterium tuberculosis
- MmpL3
- EthR2
- EthA2
- BM212
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The Mycobacterium tuberculosis Cell Envelope: unravelling complex cell wall assembly, degradation and re-cycling pathways
Besra, D., Bhatt, A., Futterer, K., Alderwick, L. & Zhang, J.
1/03/19 → 28/02/25
Project: Research Councils
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MICA: Addressing the burgeoning problem of tuberculosis: Exploiting phenotypic hits to identify new protein targets for drug discovery
Besra, D., Cox, L. & Futterer, K.
1/04/18 → 31/03/22
Project: Research Councils