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Assay Methods for the Glycosyltransferases Involved in Synthesis of Bacterial Polysaccharides

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Bacterial Polysaccharides

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1954))

Abstract

Glycans play many important roles in bacterial biology and the complexity of the glycan structures requires biochemical assays in place to help characterize the biosynthetic pathways. Our focus has been on the use of enzymes from pathogens which make molecular mimics of host glycans. We have been examining glycosyltransferases that make strategic linkages in biologically active glycans which can be also exploited for potential therapeutic glycoconjugate synthesis. This chapter will provide details on assays for a variety of bacterial glycosyltransferases that we and others have used for the characterization of pathogen glycoconjugate biosynthetic pathways, and for the in vitro synthesis of human-like glycans produced by bacterial pathogens. The methods presented here should enable other assays to be developed for new pathway characterization.

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Acknowledgements

We thank Dr. Hong-Ming Chen, University of British Columbia, for the gift of BODIPY-NHS. We thank Sussex Research Chemicals, for the gift of azido-sugars. The work was supported by grants to WW from GlycoNet doi: https://doi.org/10.13039/501100009056, and BJ was supported by an Erwin Schrödinger Fellowship.

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Authors and Affiliations

  1. Department of Chemistry and Biology, Ryerson University, Toronto, ON, Canada

    Tasnim Abukar, Nakita Buenbrazo, Bettina Janesch, Laura Kell & Warren Wakarchuk

Authors
  1. Tasnim Abukar
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  2. Nakita Buenbrazo
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  3. Bettina Janesch
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  4. Laura Kell
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  5. Warren Wakarchuk
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Corresponding author

Correspondence to Warren Wakarchuk .

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Editors and Affiliations

  1. Department of Biomedical and Molecular Sciences, Queen’s University, Kingston, ON, Canada

    Inka Brockhausen

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Abukar, T., Buenbrazo, N., Janesch, B., Kell, L., Wakarchuk, W. (2019). Assay Methods for the Glycosyltransferases Involved in Synthesis of Bacterial Polysaccharides. In: Brockhausen, I. (eds) Bacterial Polysaccharides. Methods in Molecular Biology, vol 1954. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9154-9_17

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  • DOI: https://doi.org/10.1007/978-1-4939-9154-9_17

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9153-2

  • Online ISBN: 978-1-4939-9154-9

  • eBook Packages: Springer Protocols

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