Strategy for Profiling and Structure Elucidation of Mucin-Type Oligosaccharides by Mass Spectrometry
| Autor: | Jerry L. Hedrick, LeAnn L. Lindsay, Jinhua Zhang, Carlito B. Lebrilla |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
chemistry.chemical_classification
Chromatography Fourier Analysis biology Xenopus Mucins Oligosaccharides Oligosaccharide Mass spectrometry Tandem mass spectrometry biology.organism_classification High-performance liquid chromatography Analytical Chemistry chemistry Biochemistry Exoglycosidase Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Cleave Carbohydrate Conformation Animals Structural motif Chromatography High Pressure Liquid |
| Zdroj: | Analytical Chemistry. 76:5990-6001 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/ac049666s |
| Popis: | A strategy combining accurate mass determination, tandem mass spectrometry, structure homology, and exoglycosidases is described that allows the structural characterization of mucin-type O-linked oligosaccharides. The method is used to profile with quantitation the O-linked oligosaccharide (both neutral and anionic) components of the only diploid Xenopus frog, Xenopus tropicalis. Collision-induced dissociation was used to determine connectivity, to identify previously characterized oligosaccharides, and to determine the presence of structural motifs in unknown oligosaccharides. Exoglycosidase digestion was used to identify the individual residues along with the linkages. The enzymes were also used to cleave larger oligosaccharides to smaller units that are similar to previously elucidated components. By using CID, isomeric structures were compared to determine whether they were identical. In this way, the exoglycosidases were more effectively used, and their use was minimized. A total of 35 oligosaccharides including neutral, sialylated, and sulfated were characterized in this way. The relative abundances of all components were also determined based on HPLC. |
| Databáze: | OpenAIRE |
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