Microsequencing of glycans using 2-aminobenzamide and MALDI-TOF mass spectrometry: occurrence of unique linkage-dependent fragmentation.
2-Aminobenzamide-derivatized oligosaccharides were separated by three lectin column chromatographies and then subjected to matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) for structural characterization of the carbohydrates. The combination of sequential exoglycosidase digestion and MALDI-TOF MS greatly facilitates the monosaccharide sequencing and is more feasible than size-exclusion column chromatography in terms of the time consumed and the laboriousness of the procedure. By this strategy, microsequencing of 2-3 pmol of oligosaccharide derivatives could be achieved. Furthermore, spectra obtained by the post source decay (PSD) mode provide excellent sequence information. The relative intensities of metastable ions due to fragmentation at glycosidic linkages were different among linkage isomers of particular oligosaccharides. These results demonstrate that PSD analysis possesses significant potential for the estimation of glycosidic linkage in carbohydrate structures.
Sato
Y
Y
Department of Glycobiology, Tokyo Metropolitan Institute of Gerontology, Japan.
Suzuki
M
M
Nirasawa
T
T
Suzuki
A
A
Endo
T
T
D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D062367:ortho-Aminobenzoates / Q000737:chemistry*
D011134:Polysaccharides; D062367:ortho-Aminobenzoates; C000219:anthranilamide