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Biochemical characterisation of the chlamydial MurF ligase, and possible sequence of the chlamydial peptidoglycan pentapeptide stem

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Abstract

Chlamydiaceae are obligate intracellular bacteria that do not synthesise detectable peptidoglycan although they possess an almost complete arsenal of genes encoding peptidoglycan biosynthetic activities. In this paper, the murF gene from Chlamydia trachomatis was shown to be capable of complementing a conditional Escherichia coli mutant impaired in UDP-MurNAc-tripeptide:d-Ala-d-Ala ligase activity. Recombinant MurF from C. trachomatis was overproduced and purified from E. coli. It exhibited ATP-dependent UDP-MurNAc-X-γ-d-Glu-meso-A2pm:d-Ala-d-Ala ligase activity in vitro. No significant difference of kinetic parameters was seen when X was l-Ala, l-Ser or Gly. The l-Lys-containing UDP-MurNAc-tripeptide was a poorer substrate as compared to the meso-A2pm-containing one. Based on the respective substrate specificities of the chlamydial MurC, MurE, MurF and Ddl enzymes, a sequence l-Ala/l-Ser/Gly-γ-d-Glu-meso-A2pm-d-Ala-d-Ala is expected for the chlamydial pentapeptide stem, with Gly at position 1 being less likely.

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Acknowledgments

This work was supported by grants from the Centre National de la Recherche Scientifique (UMR 8619 and PICS 2950) and the European Union EUR-INTAFAR project (LSHM-CT-2004512138).

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Authors and Affiliations

  1. Laboratoire des Enveloppes Bactériennes et Antibiotiques, Centre National de la Recherche Scientifique, Institut de Biochimie et Biophysique Moléculaire et Cellulaire, UMR 8619, Bâtiment 430, Université Paris-Sud, 91405, Orsay Cedex, France

    Delphine Patin, Dominique Mengin-Lecreulx & Didier Blanot

  2. Antimicrobial Research Centre and Institute of Molecular and Cellular Biology, University of Leeds, Leeds, LS2 9JT, UK

    Julieanne Bostock & Ian Chopra

Authors
  1. Delphine Patin
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  2. Julieanne Bostock
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  3. Ian Chopra
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  4. Dominique Mengin-Lecreulx
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  5. Didier Blanot
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Correspondence to Didier Blanot.

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Communicated by Jean-Luc Pernodet.

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Patin, D., Bostock, J., Chopra, I. et al. Biochemical characterisation of the chlamydial MurF ligase, and possible sequence of the chlamydial peptidoglycan pentapeptide stem. Arch Microbiol 194, 505–512 (2012). https://doi.org/10.1007/s00203-011-0784-8

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