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Structural and biochemical analyses of the tetrameric cell binding domain of Lys170 from enterococcal phage F170/08

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Abstract

Lysins are a class of hydrolytic enzymes used by bacteriophages to target and cleave the peptidoglycan of bacterial cell walls during their lytic cycle. The lysins from bacteriophages that infect Gram-positive bacteria are typically monomeric and consist of one or two catalytic domains (CD) and a cell binding domain (CBD). However, multimeric lysins encoded by a single gene have also been reported, among which Lys170 from enterococcal phage F170/08 was one of the first identified. Here, we determined the crystal structure of Lys170 CBD at 1.40 Å resolution. The structure reveals that Lys170 CBDs assemble into a tetrameric functional unit and that each monomer folds into a three-stranded β-sheet core capped on each side by an α-helix. In addition, we identified key residues of Lys170 CBD involved in host cell binding. Our work provides a basis for designing highly efficient lysins targeting Enterococcus faecalis.

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Availability of data and material

Atomic coordinate and structure have been deposited in the Protein Data Bank (PDB) under accession number 7D55.

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Acknowledgements

The authors thank the staff at beamline BL17U1 of Shanghai Synchrotron of Radiation Facility (SSRF) for their assistance with diffraction data collection. We thank Zhimin Guo from The First Hospital, Jilin University, Changchun, China for providing the strains of Enterococcus faecalis. We also thank our colleague Vanja Perčulija for language editing and improving the readability of the article.

Funding

This work was supported by the National Nature Science Foundation of China grants (31770948 and 31800159), the Special Open Fund of Key Laboratory of Experimental Marine Biology, Chinese Academy of Sciences (SKF2020NO1), Marine Economic Development Special Fund of Fujian Province (FJHJF-L-2020-2), the project of University-Industry Cooperation from Fujian Provincial Department of Science and Technology (2020Y4007), Natural Science Foundation of Fujian Province (2019J05065), High-level personnel introduction grant of Fujian Normal University (Z0210509), and the Fujian Provincial Department of Science and Technology (2020Y4007, 2021H0004).

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Author notes

  1. Xiaolong Xu and Dandan Zhang contributed equally to this work.

Authors and Affiliations

  1. The Key Laboratory of Innate Immune Biology of Fujian Province, Provincial University Key Laboratory of Cellular Stress Response and Metabolic Regulation, Biomedical Research Center of South China, Key Laboratory of OptoElectronic Science and Technology for Medicine of the Ministry of Education, College of Life Sciences, Fujian Normal University, Fuzhou, China

    Xiaolong Xu, Dandan Zhang, Biao Zhou, Xiangkai Zhen & Songying Ouyang

  2. The Engineering Technology Research Center of Characteristic Medicinal Plants of Fujian, Ningde Normal University, Ningde, China

    Dandan Zhang

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  1. Xiaolong Xu
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Contributions

Songying Ouyang and Xiangkai Zhen conceived and designed the experiments. Xiaolong Xu and Dandan Zhang, Biao Zhou expressed Lys170 protein and Lys170 CBD mutants, performed crystal screening, crystal optimization, data collection, bactericidal activity assay, Xiangkai Zhen determined the structure of Lys170 CBD. Xiaolong Xu, Xiangkai Zhen and Songying Ouyang analyzed the data, Xiangkai Zhen and Songying Ouyang wrote the manuscript. All authors have read and approved the manuscript.

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Correspondence to Xiangkai Zhen or Songying Ouyang.

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Xu, X., Zhang, D., Zhou, B. et al. Structural and biochemical analyses of the tetrameric cell binding domain of Lys170 from enterococcal phage F170/08. Eur Biophys J 50, 721–729 (2021). https://doi.org/10.1007/s00249-021-01511-x

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  • DOI: https://doi.org/10.1007/s00249-021-01511-x

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