title: A sequential bioorthogonal dual strategy: ManNAl and SiaNAl as distinct tools to unravel sialic acid metabolic pathways.
abstract: Recent methodological developments in metabolic oligosaccharide engineering (MOE) pave the way for tremendous advances in glycobiology. Herein, we propose a Sequential Bioorthogonal Dual Strategy (SBDS) combining the use of two unprotected alkyne-tagged monosaccharide reporters (ManNAl and SiaNAl) with the bioligation of fluorescent probes by copper-catalysed azide-alkyne cycloaddition (CuAAC). With SBDS, we are able to shed light on trafficking and cellular uptake mechanisms of sialic acid. Using their corresponding analogues, we visualized that SiaNAl enters via endocytosis, whereas its biosynthetic intermediate ManNAl uptake is mediated by a yet unknown but specific plasma membrane transporter. Sialin, a lysosomal protein, is shown to be crucial for the export of exogenous sialic acid from lysosomes to the cytosol. Metabolic labeling with alkyne-tagged derivatives of N-acetylneuraminic acid (Neu5Ac) or N-acetylmannosamine (ManNAc) could thus be used to follow endocytosis in physiological vs. pathological conditions.
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: P A
authors.initials: PA
authors.lastname: Gilormini
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: C
authors.initials: C
authors.lastname: Lion
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: D
authors.initials: D
authors.lastname: Vicogne
authors.affiliation: Laboratoire de Biochimie Métabolique, IFB, CHU Purpan, INSERM UMR 1037, CRCT, Université Paul Sabatier Toulouse III, Toulouse, France.
authors.forename: T
authors.initials: T
authors.lastname: Levade
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: S
authors.initials: S
authors.lastname: Potelle
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: C
authors.initials: C
authors.lastname: Mariller
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: Y
authors.initials: Y
authors.lastname: Guérardel
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: C
authors.initials: C
authors.lastname: Biot
authors.affiliation: Univ. Lille, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F-59000 Lille, France. christophe.biot@univ-lille1.fr francois.foulquier@univ-lille1.fr and CNRS, UMR 8576, F-59000 Lille, France.
authors.forename: F
authors.initials: F
authors.lastname: Foulquier
mesh_terms: D002384:Catalysis; D003300:Copper / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry*; D012794:Sialic Acids / Q000737:chemistry*
chemical_list: D009844:Oligosaccharides; D012794:Sialic Acids; D003300:Copper