Analysis of O-linked oligosaccharide alditols by high-pH anion-exchange chromatography with pulsed amperometric detection.
To apply high-pH anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) to the accurate and sensitive analysis of O-linked oligosaccharide alditol, removal of glycopeptides and peptides from oligosaccharide samples obtained by beta-elimination was first investigated. The results obtained by using fetuin as a model glycoprotein indicated that (glyco)peptides derivatized by fluorescamine are easily removed from the beta-eliminated oligosaccharide sample by means of a Sep-Pak C18 cartridge due to their increased hydrophobicity. This clean-up procedure was also effectively applied to the beta-eliminated oligosaccharide sample obtained from delipidated sheep erythrocyte ghosts. Furthermore, structural analysis of the oligosaccharide alditols by HPAEC-PAD was successfully performed in combination with partial and complete desialylation, composition analysis, periodate oxidation, and glycosidase digestion and revealed that sheep erythrocyte membrane glycoproteins contain Gal beta 1-->3Gal-NAc and Gal beta 1-->4GlcNAc beta 1-->3Gal beta 1-->3GalNac, to which one or two residues of Neu5Ac and Neu5Gc are attached in different combinations.
Kotani
N
N
Department of Biochemistry, University of Tokyo, Japan.
Takasaki
S
S
D000818:Animals; D001894:Borohydrides / Q000737:chemistry; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004910:Erythrocyte Membrane / Q000737:chemistry; D008040:Genetic Linkage; D008562:Membrane Glycoproteins / Q000032:analysis / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000032:analysis / Q000737:chemistry; D009438:Neuraminic Acids / Q000032:analysis / Q000737:chemistry; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry*; D012756:Sheep; D013402:Sugar Alcohols / Q000737:chemistry
D001894:Borohydrides; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D009438:Neuraminic Acids; D009844:Oligosaccharides; D013402:Sugar Alcohols; C032592:N-glycolylneuraminic acid; C025364:sodium borohydride; D019158:N-Acetylneuraminic Acid