title: Calculated two-dimensional sugar map of pyridylaminated oligosaccharides: elucidation of the jack bean alpha-mannosidase digestion pathway of Man9GlcNAc2.
abstract: We have developed a two-dimensional (2-D) mapping of pyridylaminated oligosaccharides as an aid to structural determination of glycoprotein-derived oligosaccharides. Using the available data of reverse-phase HPLC of pyridylamino-oligosaccharides, this was further extended to parameterization of unit contribution by each sugar component, which allows the prediction of possible structures from the elution volume. We have extended this approach to the data obtained with amide-silica HPLC column to obtain a calculated 2-D mapping technique for the oligomannose-type oligosaccharides (M-series). In this method, the elution volumes of all possible pyridylamino-oligosaccharides up to the size of Glc1Man9GlcNAc2 (50 in total) are calculated from the established UC values to construct a 2-D map. To test the validity of the calculated 2-D map, the structures of intermediate PA-oligosaccharides generated during the alpha-mannosidase (jack bean) digestion of Man9GlcNAc2 (porcine thyroglobulin) were analyzed to establish the digestion pathway. The validity of this approach is substantiated by an independent deduction of the intermediate structures based on structural relationships and the coincidence of elution volumes. Our results agree well with the recently published digestion pathway of Man5GlcNAc2 by the same enzyme and that of Man9GlcNAc2 by lysosomal alpha-mannosidase.
authors.affiliation: Mie Research Laboratory, Sanwa Kagaku Kenkyusho Co. Ltd, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Tomiya
authors.forename: Y C
authors.initials: YC
authors.lastname: Lee
authors.forename: T
authors.initials: T
authors.lastname: Yoshida
authors.forename: Y
authors.initials: Y
authors.lastname: Wada
authors.forename: J
authors.initials: J
authors.lastname: Awaya
authors.forename: M
authors.initials: M
authors.lastname: Kurono
authors.forename: N
authors.initials: N
authors.lastname: Takahashi
mesh_terms: D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008351:Mannans / Q000378:metabolism*; D008361:Mannosidases / Q000378:metabolism*; D008745:Methylation; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009844:Oligosaccharides / Q000737:chemistry*; D011725:Pyridines / Q000737:chemistry; D043323:alpha-Mannosidase
chemical_list: D002241:Carbohydrates; D008351:Mannans; D009844:Oligosaccharides; D011725:Pyridines; C057328:mannosyl(9)-N-acetylglucosamine; D008361:Mannosidases; D043323:alpha-Mannosidase