Van Pelt J, Van Bilsen D G, Kamerling J P, Vliegenthart J F
Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
Eur J Biochem. 1988 May 16;174(1):183-7. doi: 10.1111/j.1432-1033.1988.tb14080.x.
Sialidosis urine was fractionated by gel filtration on Bio-Gel P-6. All pooled fractions containing carbohydrates showed the presence of small amounts of GalNAc in non-reducing position, besides free N-acetyllactosamine type of oligosaccharides as major constituents. The fractions were subjected to reductive alkaline borohydride degradation, after which the major part of GalNAc was recovered as N-acetyl-D-galactosaminitol (GalNAc-ol). The GalNAc-ol-containing material was separated from the N-glycosidic oligosaccharides by a second gel-filtration step on AcA 202. Subsequently, the O-glycosidic sialyloligosaccharide-alditols were subfractionated by anion-exchange chromatography on Mono Q. Structural analysis by 500-MHz 1H-NMR spectroscopy revealed two major components in all fractions, namely: NeuAc alpha 2-3Gal beta 1-3GalNAc-ol and NeuAc alpha 2-3Gal beta 1-3[NeuAc alpha 2-6]GalNAc-ol. Furthermore, NeuAc alpha 2-3Gal beta 1-3[NeuAc alpha 2-3Gal beta 1-4GlcNAc beta 1-6]GalNAc-ol was found as a minor component in some of the fractions. The presence of these carbohydrate chains in Bio-Gel fractions differing in molecular mass suggested that they are derived from glycopeptides which are heterogeneous in their peptide part.
对唾液酸沉积症患者的尿液进行Bio-Gel P-6凝胶过滤分级分离。所有含有碳水化合物的合并级分，除了以游离N-乙酰乳糖胺型寡糖为主要成分外，在非还原端均显示存在少量的N-乙酰半乳糖胺（GalNAc）。将这些级分进行还原性碱性硼氢化物降解，之后大部分GalNAc以N-乙酰-D-半乳糖胺醇（GalNAc-ol）的形式回收。通过在AcA 202上进行第二次凝胶过滤步骤，将含GalNAc-ol的物质与N-糖苷寡糖分离。随后，通过Mono Q阴离子交换色谱对O-糖苷唾液酸寡糖糖醇进行细分。通过500兆赫兹的1H-NMR光谱进行结构分析，结果显示所有级分中均存在两种主要成分，即：NeuAcα2-3Galβ1-3GalNAc-ol和NeuAcα2-3Galβ1-3[NeuAcα2-6]GalNAc-ol。此外，在某些级分中还发现NeuAcα2-3Galβ1-3[NeuAcα2-3Galβ1-4GlcNAcβ1-6]GalNAc-ol作为次要成分。这些碳水化合物链存在于分子量不同的Bio-Gel级分中，这表明它们源自肽部分具有异质性的糖肽。
