Identification of KasA as the cellular target of an anti-tubercular scaffold

Katherine A. Abrahams, Chun-Wa Chung, Sonja Ghidelli-Disse, Joaquín Rullas, María José Rebollo-López, Sudagar S. Gurcha, Jonathan A.G. Cox, Alfonso Mendoza, Elena Jiménez-Navarro, María Santos Martínez-Martínez, Margarete Neu, Anthony Shillings, Paul Homes, Argyrides Argyrou, Ruth Casanueva, Nicholas J. Loman, Patrick J. Moynihan, Joël Lelièvre, Carolyn Selenski, Matthew Axtman & 10 others Laurent Kremer, Marcus Bantscheff, Iñigo Angulo-Barturen, Mónica Cacho Izquierdo, Nicholas C. Cammack, Gerard Drewes, Lluis Ballell, David Barros, Gurdyal S. Besra, Robert H. Bates

Research output: Contribution to journalArticle

Abstract

Phenotypic screens for bactericidal compounds are starting to yield promising hits against tuberculosis. In this regard, whole-genome sequencing of spontaneous resistant mutants generated against an indazole sulfonamide (GSK3011724A) identifies several specific single-nucleotide polymorphisms in the essential Mycobacterium tuberculosis β-ketoacyl synthase (kas) A gene. Here, this genomic-based target assignment is confirmed by biochemical assays, chemical proteomics and structural resolution of a KasA-GSK3011724A complex by X-ray crystallography. Finally, M. tuberculosis GSK3011724A-resistant mutants increase the in vitro minimum inhibitory concentration and the in vivo 99% effective dose in mice, establishing in vitro and in vivo target engagement. Surprisingly, the lack of target engagement of the related β-ketoacyl synthases (FabH and KasB) suggests a different mode of inhibition when compared with other Kas inhibitors of fatty acid biosynthesis in bacteria. These results clearly identify KasA as the biological target of GSK3011724A and validate this enzyme for further drug discovery efforts against tuberculosis.

LanguageEnglish
Article number12581
Number of pages13
JournalNature Communications
Volume7
DOIs
Publication statusPublished - 1 Sep 2016

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tuberculosis
Mycobacterium tuberculosis
Scaffolds
Tuberculosis
Genes
Indazoles
X ray crystallography
X Ray Crystallography
Biosynthesis
Sulfonamides
Microbial Sensitivity Tests
Drug Discovery
Polymorphism
Proteomics
Single Nucleotide Polymorphism
Assays
Bacteria
Fatty Acids
Nucleotides
Genome

Bibliographical note

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this
article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

Funding: EU FP7 (261378); Bill & Melinda Gates Foundation (OPP1095631; and MRC (MR/K012118/1).

Keywords

  • Journal Article

Cite this

Abrahams, K. A., Chung, C-W., Ghidelli-Disse, S., Rullas, J., Rebollo-López, M. J., Gurcha, S. S., ... Bates, R. H. (2016). Identification of KasA as the cellular target of an anti-tubercular scaffold. Nature Communications, 7, [12581]. https://doi.org/10.1038/ncomms12581
Abrahams, Katherine A. ; Chung, Chun-Wa ; Ghidelli-Disse, Sonja ; Rullas, Joaquín ; Rebollo-López, María José ; Gurcha, Sudagar S. ; Cox, Jonathan A.G. ; Mendoza, Alfonso ; Jiménez-Navarro, Elena ; Martínez-Martínez, María Santos ; Neu, Margarete ; Shillings, Anthony ; Homes, Paul ; Argyrou, Argyrides ; Casanueva, Ruth ; Loman, Nicholas J. ; Moynihan, Patrick J. ; Lelièvre, Joël ; Selenski, Carolyn ; Axtman, Matthew ; Kremer, Laurent ; Bantscheff, Marcus ; Angulo-Barturen, Iñigo ; Izquierdo, Mónica Cacho ; Cammack, Nicholas C. ; Drewes, Gerard ; Ballell, Lluis ; Barros, David ; Besra, Gurdyal S. ; Bates, Robert H. / Identification of KasA as the cellular target of an anti-tubercular scaffold. In: Nature Communications. 2016 ; Vol. 7.
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Abrahams, KA, Chung, C-W, Ghidelli-Disse, S, Rullas, J, Rebollo-López, MJ, Gurcha, SS, Cox, JAG, Mendoza, A, Jiménez-Navarro, E, Martínez-Martínez, MS, Neu, M, Shillings, A, Homes, P, Argyrou, A, Casanueva, R, Loman, NJ, Moynihan, PJ, Lelièvre, J, Selenski, C, Axtman, M, Kremer, L, Bantscheff, M, Angulo-Barturen, I, Izquierdo, MC, Cammack, NC, Drewes, G, Ballell, L, Barros, D, Besra, GS & Bates, RH 2016, 'Identification of KasA as the cellular target of an anti-tubercular scaffold' Nature Communications, vol. 7, 12581. https://doi.org/10.1038/ncomms12581

Identification of KasA as the cellular target of an anti-tubercular scaffold. / Abrahams, Katherine A.; Chung, Chun-Wa; Ghidelli-Disse, Sonja; Rullas, Joaquín; Rebollo-López, María José; Gurcha, Sudagar S.; Cox, Jonathan A.G.; Mendoza, Alfonso; Jiménez-Navarro, Elena; Martínez-Martínez, María Santos; Neu, Margarete; Shillings, Anthony; Homes, Paul; Argyrou, Argyrides; Casanueva, Ruth; Loman, Nicholas J.; Moynihan, Patrick J.; Lelièvre, Joël; Selenski, Carolyn; Axtman, Matthew; Kremer, Laurent; Bantscheff, Marcus; Angulo-Barturen, Iñigo; Izquierdo, Mónica Cacho; Cammack, Nicholas C.; Drewes, Gerard; Ballell, Lluis; Barros, David; Besra, Gurdyal S.; Bates, Robert H.

In: Nature Communications, Vol. 7, 12581, 01.09.2016.

Research output: Contribution to journalArticle

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T1 - Identification of KasA as the cellular target of an anti-tubercular scaffold

AU - Abrahams, Katherine A.

AU - Chung, Chun-Wa

AU - Ghidelli-Disse, Sonja

AU - Rullas, Joaquín

AU - Rebollo-López, María José

AU - Gurcha, Sudagar S.

AU - Cox, Jonathan A.G.

AU - Mendoza, Alfonso

AU - Jiménez-Navarro, Elena

AU - Martínez-Martínez, María Santos

AU - Neu, Margarete

AU - Shillings, Anthony

AU - Homes, Paul

AU - Argyrou, Argyrides

AU - Casanueva, Ruth

AU - Loman, Nicholas J.

AU - Moynihan, Patrick J.

AU - Lelièvre, Joël

AU - Selenski, Carolyn

AU - Axtman, Matthew

AU - Kremer, Laurent

AU - Bantscheff, Marcus

AU - Angulo-Barturen, Iñigo

AU - Izquierdo, Mónica Cacho

AU - Cammack, Nicholas C.

AU - Drewes, Gerard

AU - Ballell, Lluis

AU - Barros, David

AU - Besra, Gurdyal S.

AU - Bates, Robert H.

N1 - This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ Funding: EU FP7 (261378); Bill & Melinda Gates Foundation (OPP1095631; and MRC (MR/K012118/1).

PY - 2016/9/1

Y1 - 2016/9/1

N2 - Phenotypic screens for bactericidal compounds are starting to yield promising hits against tuberculosis. In this regard, whole-genome sequencing of spontaneous resistant mutants generated against an indazole sulfonamide (GSK3011724A) identifies several specific single-nucleotide polymorphisms in the essential Mycobacterium tuberculosis β-ketoacyl synthase (kas) A gene. Here, this genomic-based target assignment is confirmed by biochemical assays, chemical proteomics and structural resolution of a KasA-GSK3011724A complex by X-ray crystallography. Finally, M. tuberculosis GSK3011724A-resistant mutants increase the in vitro minimum inhibitory concentration and the in vivo 99% effective dose in mice, establishing in vitro and in vivo target engagement. Surprisingly, the lack of target engagement of the related β-ketoacyl synthases (FabH and KasB) suggests a different mode of inhibition when compared with other Kas inhibitors of fatty acid biosynthesis in bacteria. These results clearly identify KasA as the biological target of GSK3011724A and validate this enzyme for further drug discovery efforts against tuberculosis.

AB - Phenotypic screens for bactericidal compounds are starting to yield promising hits against tuberculosis. In this regard, whole-genome sequencing of spontaneous resistant mutants generated against an indazole sulfonamide (GSK3011724A) identifies several specific single-nucleotide polymorphisms in the essential Mycobacterium tuberculosis β-ketoacyl synthase (kas) A gene. Here, this genomic-based target assignment is confirmed by biochemical assays, chemical proteomics and structural resolution of a KasA-GSK3011724A complex by X-ray crystallography. Finally, M. tuberculosis GSK3011724A-resistant mutants increase the in vitro minimum inhibitory concentration and the in vivo 99% effective dose in mice, establishing in vitro and in vivo target engagement. Surprisingly, the lack of target engagement of the related β-ketoacyl synthases (FabH and KasB) suggests a different mode of inhibition when compared with other Kas inhibitors of fatty acid biosynthesis in bacteria. These results clearly identify KasA as the biological target of GSK3011724A and validate this enzyme for further drug discovery efforts against tuberculosis.

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Abrahams KA, Chung C-W, Ghidelli-Disse S, Rullas J, Rebollo-López MJ, Gurcha SS et al. Identification of KasA as the cellular target of an anti-tubercular scaffold. Nature Communications. 2016 Sep 1;7. 12581. https://doi.org/10.1038/ncomms12581