Preparation and structural sequence analysis of heparan sulfate dodecasaccharide
- Author:
Zi-Heng ZHANG
1
Author Information
- Publication Type:Journal Article
- Keywords: Dodecasaccharide; Heparan sulfate; Heparinase
- From: Chinese Pharmaceutical Journal 2014;49(9):781-785
- CountryChina
- Language:Chinese
- Abstract: OBJECITVE: To prepare a heparan sulfate dodecasaccharide and analyze its structural sequence. METHODS: HS-derived oligosaccharide was prepared by heparinase II depolymerization. Size-uniform dodecasaccharide mixture was acquired by low pressure gel permeation chromatography on Bio-Gel P-10 column. The dodecasaccharides were further separated by strong anion-ex-change high-performance liquid chromatography (SAX-HPLC) to get a charge-uniform dodecasaccharide. The dodecasaccharide was completely hydrolyzed by heparinases I, II, and III, its disaccharides composition was analyzed by HPLC, and its structural sequence was confirmed by electrospray collision-induced dissociation mass spectrometry (ES-CID-MSn). RESULTS: One size- and charge-uniform pure dodecasaccharide was separated from HS enzymatic hydrolyzed mixture, and it was composed by δUA-GlcNAc (IV-A) and AUA-GlcNS (IV-S) with the relative ratio of 5-1. Based on the ES-CID-MSn analysis, its structural sequence was δUA-GlcNAc-GlcA-GlcNS-GlcA-GlcNAc-GlcA-GlcNAc- GlcA-GlcNAc-GlcA-GlcNAc. CONCLUSION: The dodecasaccharide was prepared from HS, its detailed structure is analyzed, and it will offer a basis for the structure-activity relationship study of HS oligosaccharide.
