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Crystal structures of g-type lysozyme from Atlantic cod shed new light on substrate binding and the catalytic mechanism

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Abstract

Crystal structures of Atlantic cod lysozyme have been solved with and without ligand bound in the active site to 1.7 and 1.9 Å resolution, respectively. The structures reveal the presence of NAG in the substrate binding sites at both sides of the catalytic Glu73, hence allowing the first crystallographic description of the goose-type (g-type) lysozyme E–G binding sites. In addition, two aspartic acid residues suggested to participate in catalysis (Asp101 and Asp90) were mutated to alanine. Muramidase activity data for two single mutants and one double mutant demonstrates that both residues are involved in catalysis, but Asp101 is the more critical of the two. The structures and activity data suggest that a water molecule is the nucleophile completing the catalytic reaction, and the roles of the aspartic acids are to ensure proper positioning of the catalytic water.

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Acknowledgments

The present study was supported by the Research Council of Norway (NRC), project number 158952 and the National Functional Genomics Program (FUGE). Provision of beamtime at the Macromolecular Crystallography Beamlines (BL14) of Berliner Elektronenspeicherring (BESSY) and the Swiss-Norwegian Beamlines (SNBL) at European Synchrotron Radiation Facility (ESRF) is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Chemistry, The Norwegian Structural Biology Centre, University of Tromsø, 9037, Tromsø, Norway

    Ronny Helland, Renate L. Larsen, Solrun Finstad & Peter Kyomuhendo

  2. Nofima Marine, P.O. Box 6122, 9291, Tromsø, Norway

    Peter Kyomuhendo

  3. Department of Marine Biotechnology, Norwegian College of Fishery Science, University of Tromsø, 9037, Tromsø, Norway

    Atle N. Larsen

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  1. Ronny Helland
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Correspondence to Ronny Helland or Atle N. Larsen.

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Helland, R., Larsen, R.L., Finstad, S. et al. Crystal structures of g-type lysozyme from Atlantic cod shed new light on substrate binding and the catalytic mechanism. Cell. Mol. Life Sci. 66, 2585–2598 (2009). https://doi.org/10.1007/s00018-009-0063-x

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