In vitro glycosylation assay for bacterial oligosaccharyltransferases

Matias A. Musumeci, Maria V. Ielmini, Mario F. Feldman

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Scopus citations

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.

Original languageEnglish
Title of host publicationGlycosyltransferases
Subtitle of host publicationMethods and Protocols
Pages161-171
Number of pages11
DOIs
StatePublished - Jul 25 2013
Externally publishedYes

Publication series

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

Keywords

  • Glycoconjugates
  • In vitro glycosylation
  • O-linked glycosylation
  • O-oligosaccharyltransferase
  • PglL from Neisseria meningitidis

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  • Cite this

    Musumeci, M. A., Ielmini, M. V., & Feldman, M. F. (2013). In vitro glycosylation assay for bacterial oligosaccharyltransferases. In Glycosyltransferases: Methods and Protocols (pp. 161-171). (Methods in Molecular Biology; Vol. 1022). https://doi.org/10.1007/978-1-62703-465-4-13