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Methods for Determining Glycosyltransferase Kinetics

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Protein-Carbohydrate Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1588))

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Abstract

Glycosyltransferases are a class of biosynthetic enzymes that transfer individual activated monosaccharide units to specific acceptors. Colorimetric assays using the detection of released products such as para-nitrophenol and coupled assays for inorganic phosphate detection allow for convenient and quantifiable kinetic characterization. These techniques may be applied to determine the enzymatic activity of glycosyltransferases by indirectly measuring the transfer of nucleotide-activated donor carbohydrate units to various cognate acceptor molecules. In addition to an overview of these methods, the protocol for quantifying the glycosyltransferase activity used for the characterization of penicillin-binding proteins (PBPs) involving the transfer of lipid II to form elongated murein chains during bacterial cell wall synthesis is described herein.

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Acknowledgment

This work was supported by NSERC grant RGPIN-2014-05018 and the Faculty of Science, Wilfrid Laurier University.

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Authors and Affiliations

  1. Department of Chemistry & Biochemistry, Wilfrid Laurier University, 75 University Avenue West, Waterloo, ON, Canada, N2L 3C5

    Maria Ngo & Michael D. L. Suits

Authors
  1. Maria Ngo
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  2. Michael D. L. Suits
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Correspondence to Michael D. L. Suits .

Editor information

Editors and Affiliations

  1. Agriculture and Agri-Food Canada, Lethbridge, Alberta, Canada

    D. Wade Abbott

  2. Biotechnology Institute, University of Groningen, Groningen, The Netherlands

    Alicia Lammerts van Bueren

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Ngo, M., Suits, M.D.L. (2017). Methods for Determining Glycosyltransferase Kinetics. In: Abbott, D., Lammerts van Bueren, A. (eds) Protein-Carbohydrate Interactions. Methods in Molecular Biology, vol 1588. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6899-2_6

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  • DOI: https://doi.org/10.1007/978-1-4939-6899-2_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6898-5

  • Online ISBN: 978-1-4939-6899-2

  • eBook Packages: Springer Protocols

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