title: An assay for α 1,6-fucosyltransferase (FUT8) activity based on the HPLC separation of a reaction product with fluorescence detection.
abstract: N-Glycans with an α-fucose unit linked to the 6-position of the innermost GlcNAc are widely distributed among the animal kingdom, from worms and insects to human. This α1,6-linked fucosyl residue, frequently referred to as a core fucose, is formed via the action of an α1,6-fucosyltransferase, the mammalian ortholog which is systematically called FUT8. In mammals, it is well known that the extent of core-fucosylation in cellular and secreted glycoproteins varies, e.g., according to differentiation and carcinogenesis of the cells. This chapter describes a method for the sensitive and quantitative assay of FUT8 activity using a fluorescence-labeled oligosaccharyl asparagine derivative as the glycosyl acceptor substrate.
authors.affiliation: Division of Molecular Cell Biology, Department of Biomolecular Sciences, Faculty of Medicine, Saga University, Saga, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Ihara
authors.forename: Hiroki
authors.initials: H
authors.lastname: Tsukamoto
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Ikeda
mesh_terms: D000818:Animals; D001216:Asparagine / Q000031:analogs & derivatives; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D057075:Enzyme Assays / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D005647:Fucosyltransferases / Q000378:metabolism*; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis / Q000378:metabolism
chemical_list: D005456:Fluorescent Dyes; D009844:Oligosaccharides; D001216:Asparagine; D005647:Fucosyltransferases; C020559:Glycoprotein 6-alpha-L-fucosyltransferase