Structural and biochemical analysis of a bacterial glycosyltransferase

Fan Zhu, Ren Wu, Hua Zhang, Hui Wu

Research output: Chapter in Book/Report/Conference proceedingChapter

5 Scopus citations

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.

Original languageEnglish (US)
Title of host publicationGlycosyltransferases
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages29-39
Number of pages11
ISBN (Print)9781627034647
DOIs
StatePublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1022
ISSN (Print)1064-3745

Keywords

  • Crystallization and data refinement
  • Glycosyltransferase assays
  • Glycosyltransferases
  • Protein purification

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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