Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu 215021, China.
Suzhou Ronnsi Pharma Co., Ltd, Suzhou, Jiangsu 215125, China.
Carbohydr Polym. 2020 Jul 15;240:116337. doi: 10.1016/j.carbpol.2020.116337. Epub 2020 Apr 21.
Two bottom-up strategies, disaccharide and oligosaccharide analyses, were applied to elucidate the structure of a fucosylated chondroitin sulfate (FCS). The FCS was hydrolyzed with mild acid. The remained part was digested with CS lyase for disaccharide analysis. The products from each step were analyzed and the results revealed that mild sulfuric acid mainly released sulfated fucose branches, but also affected some residues and sulfo-groups on the backbone. Over 140 oligosaccharide fragments were generated by catalytic oxidation and identified by HPSEC-MS, including sulfated fucose oligosaccharides exclusively from branches, sulfated backbone fragments, and junctional fragments. Based on the results provided by these two methods, the proposed backbone of the FCS is mainly composed of GlcA→GalNAc4S6S and GlcA→GalNAc6S, and the branch is mainly located at GalNAc. The longest branch observed is nonasaccharide, and most of the fucose on the branches are mono and/or di-sulfated. NMR results supported the conclusion.
两种自下而上的策略，即二糖和寡糖分析，被应用于阐明一种岩藻糖基硫酸软骨素（FCS）的结构。FCS 用弱酸水解。剩余部分用 CS 裂解酶进行二糖分析。对每个步骤的产物进行分析，结果表明，温和的硫酸主要释放硫酸岩藻糖支链，但也会影响一些残基和主链上的磺酸基。通过催化氧化生成了 140 多种寡糖片段，并通过 HPSEC-MS 进行了鉴定，包括仅来自支链的硫酸岩藻糖寡糖、硫酸化的主链片段和连接片段。基于这两种方法提供的结果，提出的 FCS 主链主要由 GlcA→GalNAc4S6S 和 GlcA→GalNAc6S 组成，支链主要位于 GalNAc 上。观察到的最长支链是九糖，支链上的大多数岩藻糖是单磺酸化和/或二磺酸化的。NMR 结果支持了这一结论。
