Makino Y, Omichi K, Hase S
Department of Chemistry, Osaka University Graduate School of Science, Japan.
Anal Biochem. 1998 Nov 15;264(2):172-9. doi: 10.1006/abio.1998.2848.
A new sensitive and convenient method for the structural analysis of oligosaccharides was developed, in which partial acid hydrolysis was combined with two-dimensional sugar mapping of pyridylamino (PA)-oligosaccharides. A PA-oligosaccharide was partially hydrolyzed, the acid hydrolysis conditions being optimized so as to obtain various PA-oligosaccharide fragments with high yields from different types of PA-oligosaccharides. The acid hydrolyzates were then fractionated every 1 glucose unit by size-fractionation HPLC, and each fraction was analyzed by reversed-phase HPLC. The structure of each PA-oligosaccharide fragment was identified on a two-dimensional sugar map prepared with standard PA-sugar chains, after which the original PA-oligosaccharide was reconstructed from the reducing end terminal based on the obtained structures of the PA-oligosaccharide fragments. The method was successfully applied to resolving the structure of an N-linked sugar chain.
开发了一种用于寡糖结构分析的新的灵敏且便捷的方法，该方法将部分酸水解与吡啶氨基（PA）-寡糖的二维糖图谱相结合。对PA-寡糖进行部分水解，优化酸水解条件，以便从不同类型的PA-寡糖中高产率地获得各种PA-寡糖片段。然后通过尺寸排阻高效液相色谱法按每1个葡萄糖单元对酸水解产物进行分级分离，并用反相高效液相色谱法对每个级分进行分析。在用标准PA-糖链制备的二维糖图谱上鉴定每个PA-寡糖片段的结构，之后根据获得的PA-寡糖片段的结构从还原端末端重建原始的PA-寡糖。该方法成功应用于解析N-连接糖链的结构。
