Structural analysis of O-linked oligosaccharide-alditols by electrospray-tandem mass spectrometry after mild periodate oxidation and derivatization with 2-aminopyridine.
O-linked oligosaccharide-alditols were analyzed by a combination of high-performance liquid chromatography (HPLC) and electrospray-tandem mass spectrometry (ESI-MS/MS). First, oligosaccharide-alditols were treated with sodium meta-periodate under conditions where core N-acetylgalactosaminitol is specifically degraded. The resulting fragments were labeled with 2-aminopyridine and purified on a reversed-phase column. Pyridylamino oligosaccharides yielded protonated molecular ions in positive-ion ES-MS and gave Y-series sequence ions, arising from glycosidic cleavages, by ESI-tandem mass spectrometry. Information on sugar sequence and branching of oligosaccharides linked at C6 and C3 to the N-acetylgalactosaminitol can be obtained. A systematic study of various oligosaccharide-alditols demonstrated that this approach constitutes a powerful tool for the structural characterization of O-glycans available only in limited quantities.
Morelle
W
W
Laboratoire de Chimie Biologique, Université des Sciences et Technologies de Lille, Villeneuve d'Ascq, France.
Lemoine
J
J
Strecker
G
G
D000631:Aminopyridines; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D007202:Indicators and Reagents; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D010084:Oxidation-Reduction; D010504:Periodic Acid; D018629:Spectrometry, Mass, Secondary Ion / Q000379:methods; D013402:Sugar Alcohols; D014312:Trisaccharides / Q000737:chemistry
D000631:Aminopyridines; D007202:Indicators and Reagents; D009844:Oligosaccharides; D013402:Sugar Alcohols; D014312:Trisaccharides; D010504:Periodic Acid; C032439:alpha-aminopyridine