title: In vitro glycosylation assay for bacterial oligosaccharyltransferases.
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.
authors.affiliation: Department of Biological Sciences, Alberta Glycomics Centre, University of Alberta, Edmonton, AB, Canada.
authors.forename: Matias A
authors.initials: MA
authors.lastname: Musumeci
authors.forename: Maria V
authors.initials: MV
authors.lastname: Ielmini
authors.forename: Mario F
authors.initials: MF
authors.lastname: Feldman
mesh_terms: D001419:Bacteria / Q000201:enzymology*; D057075:Enzyme Assays / Q000379:methods*; D006031:Glycosylation; D006602:Hexosyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D008565:Membrane Proteins; D006602:Hexosyltransferases; C025620:dolichyl-diphosphooligosaccharide - protein glycotransferase