OBJECITVE To prepare a heparan sulfate dodecasaccharide and analyze its structural sequence.METHODS HSderived 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-exchange 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 △UA-GlcNAc (Ⅳ-A) and △UA-GlcNS(Ⅳ-S) with the relative ratio of 5∶ 1.Based on the ES-CID-MSn analysis,its structural sequence was △UAGlcNAc-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.
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万方数据
维普
