title: Tagging Glycoproteins with Fluorescently Labeled GDP-Fucoses by Using α1,3-Fucosyltransferases.
abstract: Fucose-containing glycans mediate a variety of biological processes, but there is little information on reaction processes and mechanisms mediated by fucosyltransferases. We recently reported on fluorescently labeled GDP-β-L-fucose-ATTO 550, which enabled monitoring of α1,3-fucosyltransferase activity. Here we present an extension to the previously described results, based on the synthesis of a fluorescein-isothiocyanate (FITC)-labeled and two carboxyfluorescein-labeled (FAM-labeled) NDP-β-L-fucose derivatives, and applied all four compounds in labeling of different glycoproteins with the aid of four different fucosyltransferases. The labeling processes were analyzed by in-gel fluorescence and fluorescence polarization measurements. Comparison with the ATTO-labeled sugar revealed that the FITC-labeled fucose was the best of these substrates, and that the bacterial enzyme HP-FucT tolerated the fluorescent substrates better than human fucosyltransferases.
authors.affiliation: Biochemistry, Department of Chemistry, Faculty of Sciences, Hamburg University, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany).
authors.forename: Katrin
authors.initials: K
authors.lastname: Seelhorst
authors.affiliation: Organic Chemistry, Department of Chemistry, Faculty of Sciences, Hamburg University, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany).
authors.forename: Katharina
authors.initials: K
authors.lastname: Pahnke
authors.affiliation: Organic Chemistry, Department of Chemistry, Faculty of Sciences, Hamburg University, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany).
authors.forename: Chris
authors.initials: C
authors.lastname: Meier
authors.affiliation: Biochemistry, Department of Chemistry, Faculty of Sciences, Hamburg University, Martin-Luther-King-Platz 6, 20146 Hamburg (Germany).
authors.forename: Ulrich
authors.initials: U
authors.lastname: Hahn
keywords: carbohydrates; cycloSal-nucleotides; fluorescence labeling; fucosyltransferases; glycoproteins