Yang H O, Gunay N S, Toida T, Kuberan B, Yu G, Kim Y S, Linhardt R J
Department of Chemistry, Division of Medicinal and Natural Products Chemistry, University of Iowa, Iowa City, IA 52242, USA.
Glycobiology. 2000 Oct;10(10):1033-9. doi: 10.1093/glycob/10.10.1033.
Eight oligosaccharides were prepared from dermatan sulfate (DS) and their structures were elucidated. Porcine intestinal mucosal DS was subjected to controlled depolymerization using chondroitin ABC lyase (chondroitinase ABC). The oligosaccharide mixture formed was fractionated by low-pressure gel permeation chromatography (GPC). Size uniform mixtures of disaccharides, tetrasaccharides, hexasaccharides, octasaccharides, decasaccharides, and dodecasaccharides were obtained. Each size-fractionated mixture was then purified on the basis of charge by repetitive semi-preparative strong-anion-exchange (SAX) high-performance liquid chromatography (HPLC). This approach has led to the isolation of six homogeneous oligosaccharides. The size of the oligosaccharides were determined using GPC-HPLC. Treatment of tetrasaccharide and hexasaccharide fragments with Hg(OAc)2 afforded trisaccharide and pentasaccharide products, respectively. The purity of the oligosaccharides obtained was confirmed by analytical SAX-HPLC, and capillary electrophoresis (CE). The molecular mass and degree of sulfation of the eight purified oligosaccharides were elucidated using electrospray ionization (ESI) mass spectrometry and their structures were established with high field nuclear magnetic resonance (NMR) spectroscopy. These DS-oligosaccharides are currently being used to study for interaction of the DS with biologically important proteins.
从硫酸皮肤素（DS）制备了8种寡糖，并阐明了它们的结构。使用软骨素ABC裂解酶（软骨素酶ABC）对猪肠黏膜DS进行可控解聚。形成的寡糖混合物通过低压凝胶渗透色谱法（GPC）进行分离。获得了二糖、四糖、六糖、八糖、十糖和十二糖的大小均匀混合物。然后通过重复的半制备强阴离子交换（SAX）高效液相色谱法（HPLC），根据电荷对每个大小分级的混合物进行纯化。这种方法已导致分离出6种均一的寡糖。使用GPC-HPLC测定寡糖的大小。用Hg（OAc）2处理四糖和六糖片段分别得到三糖和五糖产物。通过分析型SAX-HPLC和毛细管电泳（CE）确认所得寡糖的纯度。使用电喷雾电离（ESI）质谱法阐明了8种纯化寡糖的分子量和硫酸化程度，并通过高场核磁共振（NMR）光谱确定了它们的结构。这些DS-寡糖目前正用于研究DS与具有生物学重要性的蛋白质之间的相互作用。
