Structural determination of the O-linked sialyl oligosaccharides liberated from fetuin with endo-alpha-N-acetylgalactosaminidase-S by HPLC analysis and 600-MHz 1H-NMR spectroscopy.
The endo-alpha-N-acetylgalactosaminidase from the culture medium of Streptomyces sp. OH-11242 (endo-GalNAc-ase-S) hydrolyzed the O-glycosidic linkage between GalNAc and Ser (Thr) in fetuin, liberating oligosaccharides. The O-linked oligosaccharides liberated from the fetuin with endo-GalNAc-ase-S were pyridylaminated following fractionation on a Bio-Gel P-4 column. The structure of the pyridylaminated O-linked oligosaccharides from fetuin has been determined by reverse-phase HPLC and 600-MHz 1H-NMR spectroscopy. The chemical shifts and the coupling constants of pyridylaminated (PA) NeuAc alpha2-3Gal beta1-3GalNAc were refined by computer simulation of the spectrum. The structures of NeuAc alpha2-3Gal beta1-3(NeuAc alpha2-6)GalNAc-PA and NeuAc alpha2-3Gal beta1-3(NeuAc alpha2-3Gal beta1-4GlcNAc beta1-6)GalNAc-PA were determined by their structural reporter groups.
Ishii-Karakasa
I
I
Department of Biochemistry, School of Medicine, Kitasato University, Kanagawa, Japan.
Iwase
H
H
Hotta
K
K
D000818:Animals; D002236:Carbohydrate Conformation; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D006596:Hexosaminidases / Q000302:isolation & purification / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D009077:Mucins / Q000737:chemistry; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D013379:Substrate Specificity; D013552:Swine; D000509:alpha-Fetoproteins / Q000737:chemistry*; D048809:alpha-N-Acetylgalactosaminidase
D009077:Mucins; D009844:Oligosaccharides; D000509:alpha-Fetoproteins; C016735:sialooligosaccharides; D006596:Hexosaminidases; D048809:alpha-N-Acetylgalactosaminidase