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Crystal structure of the retaining galactosyltransferase LgtC from Neisseria meningitidis in complex with donor and acceptor sugar analogs

Nature Structural Biology volume 8, pages 166–175 (2001)Cite this article

Abstract

Many bacterial pathogens express lipooligosaccharides that mimic human cell surface glycoconjugates, enabling them to attach to host receptors and to evade the immune response. In Neisseria meningitidis, the galactosyltransferase LgtC catalyzes a key step in the biosynthesis of lipooligosaccharide structure by transferring α-d-galactose from UDP-galactose to a terminal lactose. The product retains the configuration of the donor sugar glycosidic bond; LgtC is thus a retaining glycosyltranferase. We report the 2 Å crystal structures of the complex of LgtC with manganese and UDP 2-deoxy-2-fluoro-galactose (a donor sugar analog) in the presence and absence of the acceptor sugar analog 4′-deoxylactose. The structures, together with results from site-directed mutagenesis and kinetic analysis, give valuable insights into the unique catalytic mechanism and, as the first structure of a glycosyltransferase in complex with both the donor and acceptor sugars, provide a starting point for inhibitor design.

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Figure 1: Glycosyl transfer reactions.
Figure 2: An amino acid sequence alignment of N. meningitidis LgtC and related enzymes from glycosyl transferase family 8.
Figure 3: The overall architecture of LgtC.
Figure 4: The active site.
Figure 5: Schematic representation of the interactions between the enzyme and the substrate analogs.
Figure 6: Possible mechanisms of the LgtC catalyzed reaction.

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Acknowledgements

We thank H. Bellamy and the staff of the SSRL for access to beamline 1-5 for data collection. We thank D. Dombroski for performing initial crystallization trials on LgtC. Financial support from the Natural Sciences and Engineering Research Council of Canada in terms of a scholarship to H.D.L. and a strategic grant to S.G.W., W.W.W. and N.C.J.S. is gratefully acknowledged. K.P. is a Swedish Research Council for Engineering Sciences Scholar. N.C.J.S. is a MRC Scholar, Burroughs Wellcome New Investigator and Howard Hughes International Scholar.

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  1. Department of Biochemistry and Molecular Biology, University of British Columbia, 2146 Health Sciences Mall, Vancouver, V6T 1Z3, British Columbia, Canada

    Karina Persson & Natalie C.J. Strynadka

  2. Department of Chemistry, University of British Columbia, 2036 Main Mall, Vancouver, V6T 1Z1, British Columbia, Canada

    Hoa D. Ly & Stephen G. Withers

  3. Institute for Biological Sciences, National Research Council, Room 3157, 100 Sussex Drive, Ottawa, K1A OR6, Ontario, Canada

    Manuela Dieckelmann & Warren W. Wakarchuk

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Correspondence to Natalie C.J. Strynadka.

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Persson, K., Ly, H., Dieckelmann, M. et al. Crystal structure of the retaining galactosyltransferase LgtC from Neisseria meningitidis in complex with donor and acceptor sugar analogs. Nat Struct Mol Biol 8, 166–175 (2001). https://doi.org/10.1038/84168

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