The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation

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 numer...

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Autores principales:	Alice R. Moorey, Alejandro Cabanillas, Sarah M. Batt, Sonja Ghidelli-Disse, Beatriz Urones, Olalla Sanz, Joel Lelievre, Marcus Bantscheff, Liam R. Cox, Gurdyal S. Besra
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Mycobacterium tuberculosis MmpL3 EthR2 EthA2 BM212 Cytology QH573-671
Acceso en línea:	https://doaj.org/article/a33fd1f698b0424ca7a367765fd2c89c
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spelling	oai:doaj.org-article:a33fd1f698b0424ca7a367765fd2c89c2021-11-28T04:36:08ZThe multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation2468-233010.1016/j.tcsw.2021.100068https://doaj.org/article/a33fd1f698b0424ca7a367765fd2c89c2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2468233021000219https://doaj.org/toc/2468-2330The 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.Alice R. MooreyAlejandro CabanillasSarah M. BattSonja Ghidelli-DisseBeatriz UronesOlalla SanzJoel LelievreMarcus BantscheffLiam R. CoxGurdyal S. BesraElsevierarticleMycobacterium tuberculosisMmpL3EthR2EthA2BM212CytologyQH573-671ENThe Cell Surface, Vol 7, Iss , Pp 100068- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Mycobacterium tuberculosis MmpL3 EthR2 EthA2 BM212 Cytology QH573-671
spellingShingle	Mycobacterium tuberculosis MmpL3 EthR2 EthA2 BM212 Cytology QH573-671 Alice R. Moorey Alejandro Cabanillas Sarah M. Batt Sonja Ghidelli-Disse Beatriz Urones Olalla Sanz Joel Lelievre Marcus Bantscheff Liam R. Cox Gurdyal S. Besra The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation
description	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.
format	article
author	Alice R. Moorey Alejandro Cabanillas Sarah M. Batt Sonja Ghidelli-Disse Beatriz Urones Olalla Sanz Joel Lelievre Marcus Bantscheff Liam R. Cox Gurdyal S. Besra
author_facet	Alice R. Moorey Alejandro Cabanillas Sarah M. Batt Sonja Ghidelli-Disse Beatriz Urones Olalla Sanz Joel Lelievre Marcus Bantscheff Liam R. Cox Gurdyal S. Besra
author_sort	Alice R. Moorey
title	The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation
title_short	The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation
title_full	The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation
title_fullStr	The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation
title_full_unstemmed	The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation
title_sort	multi-target aspect of an mmpl3 inhibitor: the bm212 series of compounds bind ethr2, a transcriptional regulator of ethionamide activation
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/a33fd1f698b0424ca7a367765fd2c89c
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The multi-target aspect of an MmpL3 inhibitor: The BM212 series of compounds bind EthR2, a transcriptional regulator of ethionamide activation

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