Glycan profiling of urine, amniotic fluid and ascitic fluid from galactosialidosis patients reveals novel oligosaccharides with reducing end hexose and aldohexonic acid residues.
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.
Bruggink
Cees
C
Department of Parasitology, Leiden University Medical Center, Leiden, The Netherlands. c.bruggink@lumc.nl
Poorthuis
Ben J H M
BJ
Piraud
Monique
M
Froissart
Roseline
R
Deelder
André M
AM
Wuhrer
Manfred
M
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*
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