title: Glycan profiling of urine, amniotic fluid and ascitic fluid from galactosialidosis patients reveals novel oligosaccharides with reducing end hexose and aldohexonic acid residues.
abstract: Urine, amniotic fluid and ascitic fluid samples of galactosialidosis patients were analyzed and structurally characterized for free oligosaccharides using capillary high-performance anion-exchange chromatography with pulsed amperometric detection and online mass spectrometry. In addition to the expected endo-beta-N-acetylglucosaminidase-cleaved products of complex-type sialylated N-glycans, O-sulfated oligosaccharide moieties were detected. Moreover, novel carbohydrate moieties with reducing-end hexose residues were detected. On the basis of structural features such as a hexose-N-acetylhexosamine-hexose-hexose consensus sequence and di-sialic acid units, these oligosaccharides are thought to represent, at least in part, glycan moieties of glycosphingolipids. In addition, C(1)-oxidized, aldohexonic acid-containing versions of most of these oligosaccharides were observed. These observations suggest an alternative catabolism of glycosphingolipids in galactosialidosis patients: oligosaccharide moieties from glycosphingolipids would be released by a hitherto unknown ceramide glycanase activity. The results show the potential and versatility of the analytical approach for structural characterization of oligosaccharides in various body fluids.
authors.affiliation: Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands. c.bruggink@lumc.nl
authors.forename: Cees
authors.initials: C
authors.lastname: Bruggink
authors.forename: Ben J H M
authors.initials: BJ
authors.lastname: Poorthuis
authors.forename: Monique
authors.initials: M
authors.lastname: Piraud
authors.forename: Roseline
authors.initials: R
authors.lastname: Froissart
authors.forename: André M
authors.initials: AM
authors.lastname: Deelder
authors.forename: Manfred
authors.initials: M
authors.lastname: Wuhrer
mesh_terms: D000653:Amniotic Fluid / Q000737:chemistry*; D001202:Ascitic Fluid / Q000737:chemistry*; D002240:Carbohydrate Sequence; D043402:Cathepsin A / Q000235:genetics; D002852:Chromatography, Ion Exchange; D005333:Fetus / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D006028:Glycosphingolipids / Q000378:metabolism; D006601:Hexoses / Q000032:analysis; D006801:Humans; D007223:Infant; D007231:Infant, Newborn / Q000378:metabolism / Q000652:urine; D016464:Lysosomal Storage Diseases / Q000235:genetics / Q000378:metabolism* / Q000652:urine*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D009439:Neuraminidase / Q000172:deficiency*; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry / Q000652:urine; D025942:Oligosaccharides, Branched-Chain / Q000032:analysis / Q000737:chemistry / Q000652:urine; D013400:Sugar Acids / Q000032:analysis; D014556:Urine / Q000737:chemistry; D001616:beta-Galactosidase / Q000172:deficiency*
chemical_list: D006028:Glycosphingolipids; D006601:Hexoses; D009844:Oligosaccharides; D025942:Oligosaccharides, Branched-Chain; D013400:Sugar Acids; D006026:Glycoside Hydrolases; C051111:ceramide glycanase; D009439:Neuraminidase; D001616:beta-Galactosidase; D043402:Cathepsin A