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Structural analysis of the sialyltransferase CstII from Campylobacter jejuni in complex with a substrate analog

Nature Structural & Molecular Biology volume 11, pages 163–170 (2004)Cite this article

Abstract

Sialic acid terminates oligosaccharide chains on mammalian and microbial cell surfaces, playing critical roles in recognition and adherence. The enzymes that transfer the sialic acid moiety from cytidine-5′-monophospho-N-acetyl-neuraminic acid (CMP-NeuAc) to the terminal positions of these key glycoconjugates are known as sialyltransferases. Despite their important biological roles, little is understood about the mechanism or molecular structure of these membrane-associated enzymes. We report the first structure of a sialyltransferase, that of CstII from Campylobacter jejuni, a highly prevalent foodborne pathogen. Our structural, mutagenesis and kinetic data provide support for a novel mode of substrate binding and glycosyl transfer mechanism, including essential roles of a histidine (general base) and two tyrosine residues (coordination of the phosphate leaving group). This work provides a framework for understanding the activity of several sialyltransferases, from bacterial to human, and for the structure-based design of specific inhibitors.

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Figure 1: Reaction scheme of CstII.
Figure 2: The overall architecture of CstIIΔ32.
Figure 3: The active site of CstIIΔ32.
Figure 4: The donor substrate analog CMP-3FNeuAc.

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Acknowledgements

We thank the US Department of Energy for access to data collection facilities at the NSLS, and also M. Karwaski and S. Ryan for technical help. We thank Neose Pharmaceuticals for providing sialyllactose and CMP-NeuAc for the kinetic studies. The work was funded by the Howard Hughes Medical Institute, the Canadian Institute of Health Research and the Burroughs Wellcome Foundation (to N.S.), the Natural Sciences and Engineering Research Council and a Human Frontiers Science Grant (to S.W.) and by the National Research Council–GH (to W.W. and M.G.).

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  1. Andrew G Watts and Luke L Lairson: These authors contributed equally to this work.

Authors and Affiliations

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

    Cecilia P C Chiu, Daniel Lim & Natalie C J Strynadka

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

    Andrew G Watts, Luke L Lairson & Stephen G Withers

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

    Michel Gilbert & Warren W Wakarchuk

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Correspondence to Natalie C J Strynadka.

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Chiu, C., Watts, A., Lairson, L. et al. Structural analysis of the sialyltransferase CstII from Campylobacter jejuni in complex with a substrate analog. Nat Struct Mol Biol 11, 163–170 (2004). https://doi.org/10.1038/nsmb720

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