Abstract
Oligosaccharyltransferases (OTases) constitute a family of glycosyltransferases that catalyze the transfer of an oligosaccharide from a lipid donor to an acceptor molecule, commonly a protein. These enzymes can transfer a variety of glycan structures, including polysaccharides, to different protein acceptors. Therefore, this property endows the OTases with great biotechnological potential as these enzymes could be applied to produce several glycoconjugates relevant to the pharmaceutical industry. Furthermore, bacterial OTases are thought to be involved in pathogenesis mechanisms. Here we describe how to purify a representative OTase and its protein acceptor and glycan donor to perform in vitro glycosylation studies.
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Acknowledgments
We thank Ken McMahon and Nancy Price for proofreading the manuscript. This work was supported by funds from the Alberta Glycomics Centre to MFF. MFF is a CIHR New investigator and an AHFMR scholar.
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Musumeci, M.A., Ielmini, M.V., Feldman, M.F. (2013). In Vitro Glycosylation Assay for Bacterial Oligosaccharyltransferases. In: Brockhausen, I. (eds) Glycosyltransferases. Methods in Molecular Biology, vol 1022. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-465-4_13
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DOI: https://doi.org/10.1007/978-1-62703-465-4_13
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