Bactericidal mode of action of bedaquiline

Kiel Hards, Jennifer R. Robson, Michael Berney, Lisa Shaw, Dirk Bald, Anil Koul, Koen Andries, Gregory M. Cook

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Objectives: It is not fully understood why inhibiting ATP synthesis in Mycobacterium species leads to death in non-replicating cells. We investigated the bactericidal mode of action of the anti-tubercular F<inf>1</inf>F<inf>o</inf>-ATP synthase inhibitor bedaquiline (Sirturo™) in order to further understand the lethality of ATP synthase inhibition. Methods: Mycobacterium smegmatis strains were used for all the experiments. Growth and survival during a bedaquiline challenge were performed in multiple media types. A time-course microarray was performed during initial bedaquiline challenge in minimal medium. Oxygen consumption and proton-motive force measurements were performed on whole cells and inverted membrane vesicles, respectively. Results: A killing of 3 log.<inf>10</inf> cfu/mL was achieved 4-fold more quickly in minimal medium (a glycerol carbon source) versus rich medium (LB with Tween 80) during bedaquiline challenge. Assessing the accelerated killing condition, we identified a transcriptional remodelling of metabolism that was consistent with respiratory dysfunction but inconsistent with ATP depletion. In glycerol-energized cell suspensions, bedaquiline caused an immediate 2.3-fold increase in oxygen consumption. Bedaquiline collapsed the transmembrane pH gradient, but not the membrane potential, in a dose-dependent manner. Both these effects were dependent on binding to the F.<inf>1</inf>F.<inf>o</inf>-ATP synthase. Conclusions: Challenge with bedaquiline results in an electroneutral uncoupling of respiration-driven ATP synthesis. This may be a determinant of the bactericidal effects of bedaquiline, while ATP depletion may be a determinant of its delayed onset of killing. We propose that bedaquiline binds to and perturbs the a-c subunit interface of the F<inf>o</inf>, leading to futile proton cycling, which is known to be lethal to mycobacteria.

Original languageEnglish (US)
Article numberdkv054
Pages (from-to)2028-2037
Number of pages10
JournalJournal of Antimicrobial Chemotherapy
Volume70
Issue number7
DOIs
StatePublished - Dec 6 2014

Fingerprint

bedaquiline
Adenosine Triphosphate
Proton-Motive Force
Mycobacterium
Oxygen Consumption
Glycerol
Substrate Cycling
Mycobacterium smegmatis
Polysorbates
Membrane Potentials

Keywords

  • Antimycobacterial agents
  • F<inf>1</inf>F<inf>o</inf>-ATP synthase
  • Mycobacteria
  • R207910
  • TMC207

ASJC Scopus subject areas

  • Pharmacology
  • Pharmacology (medical)
  • Infectious Diseases

Cite this

Hards, K., Robson, J. R., Berney, M., Shaw, L., Bald, D., Koul, A., ... Cook, G. M. (2014). Bactericidal mode of action of bedaquiline. Journal of Antimicrobial Chemotherapy, 70(7), 2028-2037. [dkv054]. https://doi.org/10.1093/jac/dkv054

Bactericidal mode of action of bedaquiline. / Hards, Kiel; Robson, Jennifer R.; Berney, Michael; Shaw, Lisa; Bald, Dirk; Koul, Anil; Andries, Koen; Cook, Gregory M.

In: Journal of Antimicrobial Chemotherapy, Vol. 70, No. 7, dkv054, 06.12.2014, p. 2028-2037.

Research output: Contribution to journalArticle

Hards, K, Robson, JR, Berney, M, Shaw, L, Bald, D, Koul, A, Andries, K & Cook, GM 2014, 'Bactericidal mode of action of bedaquiline', Journal of Antimicrobial Chemotherapy, vol. 70, no. 7, dkv054, pp. 2028-2037. https://doi.org/10.1093/jac/dkv054
Hards K, Robson JR, Berney M, Shaw L, Bald D, Koul A et al. Bactericidal mode of action of bedaquiline. Journal of Antimicrobial Chemotherapy. 2014 Dec 6;70(7):2028-2037. dkv054. https://doi.org/10.1093/jac/dkv054
Hards, Kiel ; Robson, Jennifer R. ; Berney, Michael ; Shaw, Lisa ; Bald, Dirk ; Koul, Anil ; Andries, Koen ; Cook, Gregory M. / Bactericidal mode of action of bedaquiline. In: Journal of Antimicrobial Chemotherapy. 2014 ; Vol. 70, No. 7. pp. 2028-2037.
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abstract = "Objectives: It is not fully understood why inhibiting ATP synthesis in Mycobacterium species leads to death in non-replicating cells. We investigated the bactericidal mode of action of the anti-tubercular F1Fo-ATP synthase inhibitor bedaquiline (Sirturo™) in order to further understand the lethality of ATP synthase inhibition. Methods: Mycobacterium smegmatis strains were used for all the experiments. Growth and survival during a bedaquiline challenge were performed in multiple media types. A time-course microarray was performed during initial bedaquiline challenge in minimal medium. Oxygen consumption and proton-motive force measurements were performed on whole cells and inverted membrane vesicles, respectively. Results: A killing of 3 log.10 cfu/mL was achieved 4-fold more quickly in minimal medium (a glycerol carbon source) versus rich medium (LB with Tween 80) during bedaquiline challenge. Assessing the accelerated killing condition, we identified a transcriptional remodelling of metabolism that was consistent with respiratory dysfunction but inconsistent with ATP depletion. In glycerol-energized cell suspensions, bedaquiline caused an immediate 2.3-fold increase in oxygen consumption. Bedaquiline collapsed the transmembrane pH gradient, but not the membrane potential, in a dose-dependent manner. Both these effects were dependent on binding to the F.1F.o-ATP synthase. Conclusions: Challenge with bedaquiline results in an electroneutral uncoupling of respiration-driven ATP synthesis. This may be a determinant of the bactericidal effects of bedaquiline, while ATP depletion may be a determinant of its delayed onset of killing. We propose that bedaquiline binds to and perturbs the a-c subunit interface of the Fo, leading to futile proton cycling, which is known to be lethal to mycobacteria.",
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