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Structural basis of bacterial defense against g-type lysozyme-based innate immunity

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Abstract

Gram-negative bacteria can produce specific proteinaceous inhibitors to defend themselves against the lytic action of host lysozymes. So far, four different lysozyme inhibitor families have been identified. Here, we report the crystal structure of the Escherichia coli periplasmic lysozyme inhibitor of g-type lysozyme (PliG-Ec) in complex with Atlantic salmon g-type lysozyme (SalG) at a resolution of 0.95 Å, which is exceptionally high for a complex of two proteins. The structure reveals for the first time the mechanism of g-type lysozyme inhibition by the PliG family. The latter contains two specific conserved regions that are essential for its inhibitory activity. The inhibitory complex formation is based on a double ‘key-lock’ mechanism. The first key-lock element is formed by the insertion of two conserved PliG regions into the active site of the lysozyme. The second element is defined by a distinct pocket of PliG accommodating a lysozyme loop. Computational analysis indicates that this pocket represents a suitable site for small molecule binding, which opens an avenue for the development of novel antibacterial agents that suppress the inhibitory activity of PliG.

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Acknowledgments

Access to the synchrotron beamlines PROXIMA1 at Soleil and X33 at the Deutsches Elektronen-Synchrotron is gratefully acknowledged, with a special ‘thank you’ to Dr. Andrew Thompson for help with ultrahigh-resolution data collection. We also thank Dr. Pavel Afonine for advice on crystallographic refinement. S. Leysen holds a doctoral grant from the K.U. Leuven. L. Vanderkelen holds a doctoral fellowship from the Flemish Institute for the Promotion of Scientific Technological Research (IWT). This work was further financially supported by a Research Grant (G.0363.08) from the Research Foundation-Flanders (F.W.O.-Vlaanderen) and from the KU Leuven Research Fund (research project METH/07/03).

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Authors and Affiliations

  1. Laboratory for Biocrystallography, Department of Pharmaceutical and Pharmacological Sciences, Katholieke Universiteit Leuven, Herestraat 49 bus 822, 3000, Leuven, Belgium

    S. Leysen, S. D. Weeks & S. V. Strelkov

  2. Laboratory of Food Microbiology, Leuven Food Science and Nutrition Research Centre (LFoRCe), Katholieke Universiteit Leuven, Kasteelpark Arenberg 22, 3001, Leuven, Belgium

    L. Vanderkelen & C. W. Michiels

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  1. S. Leysen
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  5. S. V. Strelkov
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Correspondence to S. V. Strelkov.

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Leysen, S., Vanderkelen, L., Weeks, S.D. et al. Structural basis of bacterial defense against g-type lysozyme-based innate immunity. Cell. Mol. Life Sci. 70, 1113–1122 (2013). https://doi.org/10.1007/s00018-012-1184-1

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