Abstract
Phages infecting the polysialic acid (polySia)-encapsulated human pathogen Escherichia coli K1 are equipped with capsule-degrading tailspikes known as endosialidases, which are the only identified enzymes that specifically degrade polySia. As polySia also promotes cellular plasticity and tumor metastasis in vertebrates, endosialidases are widely applied in polySia-related neurosciences and cancer research. Here we report the crystal structures of endosialidase NF and its complex with oligomeric sialic acid. The structure NF, which reveals three distinct domains, indicates that the unique polySia specificity evolved from a combination of structural elements characteristic of exosialidases and bacteriophage tailspike proteins. The endosialidase assembles into a catalytic trimer stabilized by a triple β-helix. Its active site differs markedly from that of exosialidases, indicating an endosialidase-specific substrate-binding mode and catalytic mechanism. Residues essential for endosialidase activity were identified by structure-based mutational analysis.
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Acknowledgements
We thank the staff of European Molecular Biology Laboratory beamline at DESY, Hamburg, and the staff of the PSF beamline at BESSY, Berlin, for guidance during data collection. We are grateful to our colleague O. Einsle for help during the crystal structure analysis. This work was supported by the Deutsche Forschungsgemeinschaft and the Fonds der Chemischen Industrie.
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Supplementary Fig. 1
Binding of inactive endoNF mutants to polySia. (PDF 159 kb)
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Stummeyer, K., Dickmanns, A., Mühlenhoff, M. et al. Crystal structure of the polysialic acid–degrading endosialidase of bacteriophage K1F. Nat Struct Mol Biol 12, 90–96 (2005). https://doi.org/10.1038/nsmb874
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DOI: https://doi.org/10.1038/nsmb874
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