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The Mycobacterium tuberculosis protein LdtMt2 is a nonclassical transpeptidase required for virulence and resistance to amoxicillin

Nature Medicine volume 16, pages 466–469 (2010)Cite this article

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Abstract

The peptidoglycan layer is a vital component of the bacterial cell wall. The existing paradigm describes the peptidoglycan network as a static structure that is cross-linked predominantly by 4→3 transpeptide linkages. However, the nonclassical 3→3 linkages predominate the transpeptide networking of the peptidoglycan layer of nonreplicating Mycobacterium tuberculosis1,2. The molecular basis of these linkages and their role in the physiology of the peptidoglycan layer, virulence and susceptibility of M. tuberculosis to drugs remain undefined. Here we identify MT2594 as an L,D-transpeptidase that generates 3→3 linkages in M. tuberculosis. We show that the loss of this protein leads to altered colony morphology, loss of virulence and increased susceptibility to amoxicillin-clavulanate during the chronic phase of infection. This suggests that 3→3 cross-linking is vital to the physiology of the peptidoglycan layer. Although a functional homolog exists, expression of ldtMt2 is dominant throughout the growth phases of M. tuberculosis. 4→3 transpeptide linkages are targeted by one of the most widely used classes of antibacterial drugs in human clinical use today, β-lactams. Recently, meropenem-clavulanate was shown to be effective against drug-resistant M. tuberculosis3. Our study suggests that a combination of L,D-transpeptidase and β-lactamase inhibitors could effectively target persisting bacilli during the chronic phase of tuberculosis.

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Figure 1: Morphology and growth in vitro.
Figure 2: Characterization of LdtMt2 from M. tuberculosis.
Figure 3: Assessment of growth, virulence and susceptibility to amoxicillin in vivo.
Figure 4: Proposed model for physiology of the peptidoglycan layer in M. tuberculosis.

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Acknowledgements

We gratefully acknowledge the support of US National Institutes of Health award AI30036. This work was also supported by the Foundation pour la Recherche Médicale (Equipe FRM 2006 (Dequation 200661107918)). M. Lavollay is the recipient of an Institut National de la Santé et de la Recherche Médicale PhD fellowship (Poste d'Accueil pour Pharmacien, Médecin, et Vétérinaire).

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Authors and Affiliations

  1. Department of Medicine, Johns Hopkins University, Baltimore, Maryland, USA

    Radhika Gupta, William R Bishai & Gyanu Lamichhane

  2. Centre de Recherche des Cordeliers, Laboratorie de Recherché Moleculaire sur les Antibiotiques, Université Pierre et Marie Curie, Paris, France

    Marie Lavollay, Jean-Luc Mainardi & Michel Arthur

  3. Université Paris Descartes, Paris, France

    Marie Lavollay, Jean-Luc Mainardi & Michel Arthur

  4. Institut National de la Santé et de la Recherche Médicale, Paris, France

    Marie Lavollay, Jean-Luc Mainardi & Michel Arthur

  5. Hôpital Européen Georges Pompidou, Paris, France

    Jean-Luc Mainardi

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  1. Radhika Gupta
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Contributions

R.G., W.R.B. and G.L. designed the project. J.-L.M. and M.A. designed the biochemical characterization of MT2594. M.L. and J.-L.M. performed biochemistry and analyzed data. R.G. and G.L. conducted genetics, microbiology and mouse experiments. G.L. wrote the manuscript with contributions from the other authors.

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Correspondence to Gyanu Lamichhane.

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The authors declare no competing financial interests.

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Supplementary Figs. 1–5 and Supplementary Methods (PDF 839 kb)

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Gupta, R., Lavollay, M., Mainardi, JL. et al. The Mycobacterium tuberculosis protein LdtMt2 is a nonclassical transpeptidase required for virulence and resistance to amoxicillin. Nat Med 16, 466–469 (2010). https://doi.org/10.1038/nm.2120

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