Analysis of oligosaccharide structures from the reducing end terminal by combining partial acid hydrolysis and a two-dimensional sugar map.
A new sensitive and convenient method for the structural analysis of oligosaccharides was developed, in which partial acid hydrolysis was combined with two-dimensional sugar mapping of pyridylamino (PA)-oligosaccharides. A PA-oligosaccharide was partially hydrolyzed, the acid hydrolysis conditions being optimized so as to obtain various PA-oligosaccharide fragments with high yields from different types of PA-oligosaccharides. The acid hydrolyzates were then fractionated every 1 glucose unit by size-fractionation HPLC, and each fraction was analyzed by reversed-phase HPLC. The structure of each PA-oligosaccharide fragment was identified on a two-dimensional sugar map prepared with standard PA-sugar chains, after which the original PA-oligosaccharide was reconstructed from the reducing end terminal based on the obtained structures of the PA-oligosaccharide fragments. The method was successfully applied to resolving the structure of an N-linked sugar chain.
Makino
Y
Y
Department of Chemistry, Osaka University Graduate School of Science, Japan.
Omichi
K
K
Hase
S
S
D000631:Aminopyridines / Q000378:metabolism; D000818:Animals; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D005456:Fluorescent Dyes / Q000032:analysis; D006026:Glycoside Hydrolases / Q000378:metabolism; D006868:Hydrolysis; D007136:Immunoglobulins / Q000737:chemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D012680:Sensitivity and Specificity; D016339:Spectrometry, Mass, Fast Atom Bombardment; D013696:Temperature; D013997:Time Factors; D014269:Trifluoroacetic Acid / Q000378:metabolism
D000631:Aminopyridines; D005456:Fluorescent Dyes; C016512:IgY; D007136:Immunoglobulins; D009844:Oligosaccharides; D014269:Trifluoroacetic Acid; D006026:Glycoside Hydrolases