Abstract
Glycosyltransferases (GTs) are a large family of enzymes that specifically transfer sugar moieties to a diverse range of substrates. The process of bacterial glycosylation (such as biosynthesis of glycolipids, glycoproteins, and polysaccharides) has been studied extensively, yet the majority of GTs involved remains poorly characterized. Besides predicting enzymatic parameters of GTs, the resolution of three-dimensional structures of GTs can help to determine activity, donor sugar binding, and acceptor substrate binding sites. It also facilitates amino acid sequence-based structural modeling and biochemical characterization of their homologues. Here we describe a general procedure to accomplish expression and purification of soluble and active recombinant GTs. Enzymatic characterization, and crystallization of GTs, and data refinement for structural analysis are also covered in this protocol.
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Acknowledgments
We thank Dr. Heidi Erlandsen for critical reading of the manuscript. The work was supported by NIH/NIDCR grant R01DE017954 (HW).
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Zhu, F., Wu, R., Zhang, H., Wu, H. (2013). Structural and Biochemical Analysis of a Bacterial Glycosyltransferase. 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_3
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DOI: https://doi.org/10.1007/978-1-62703-465-4_3
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Publisher Name: Humana Press, Totowa, NJ
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