Synthesis of 3-O-Sulfated Disaccharide and Tetrasaccharide Standards for Compositional Analysis of Heparan Sulfate
Synthesis of 3-O-Sulfated Disaccharide and Tetrasaccharide Standards for Compositional Analysis of Heparan Sulfate
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DOI：
10.1021/acs.biochem.9b00838
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发表时间：
2020-09-01
期刊：
影响因子：
2.9
通讯作者：
Liu, Jian
中科院分区：
文献类型：
--
作者：
Dhurandhare, Vijay Manohar;Pagadala, Vijayakanth;Liu, Jian
硫酸乙酰肝素（HS）中葡萄糖胺糖单元上的3-O-硫酸化与各种生物功能有关，包括在医院治疗血栓性疾病的抗凝活性。3-O-硫酸葡糖胺由硫酸乙酰肝素葡糖胺3-磺基转移酶生物合成。由于其生物学意义，需要3-O-硫酸化寡糖标准品以促进HS的组成分析。这些寡糖必须在非还原端含有Δ（4，5）-不饱和糖醛酸（Δ UA）残基，这是由于在组成分析程序期间使用的肝素裂解酶催化的解聚反应。在此，我们描述了以毫克规模制备一种3-O-硫酸化二糖（化合物4）和三种3-O-硫酸化四糖（化合物1-3）的方案。通过使用肝素裂解酶降解合成的八糖来完成3-O-硫酸化二糖和四糖标准品的合成。进一步分析表明，3-O-硫酸寡糖标准品在碱性条件下不稳定，证实了先前研究的结果。通过在磷酸盐缓冲液存在下降低pH值来减少不需要的降解。3-O-硫酸寡糖标准品是用于表征生物来源HS中3-O-硫酸化的试剂。
3-O-Sulfation on the glucosamine sugar unit in heparan sulfate (HS) is linked to various biological functions, including the anticoagulant activity to treat thrombotic disorders in hospitals. The 3-O-sulfated glucosamine is biosynthesized by heparan sulfate glucosamine 3-sulfotransferases. Because of its biological significance, there is a need for 3-O-sulfated oligosaccharide standards to facilitate the compositional analysis of HS. These oligosaccharides must contain a Delta(4,5)-unsaturated uronic acid (Delta UA) residue at the nonreducing end, which is due to the depolymerization reaction catalyzed by heparin lyases used during the compositional analysis procedure. Here, we describe a protocol for the preparation of one 3-O-sulfated disaccharide (compound 4) and three 3-O-sulfated tetrasaccharides (compound 1-3) in a milligram scale. The synthesis of 3-O-sulfated disaccharide and tetrasaccharide standards was completed by degrading synthetic octasaccharides using heparin lyases. Further analysis revealed that 3-O-sulfated oligosaccharide standards are labile under basic conditions, confirming the findings from a previous study. The unwanted degradation was reduced by decreasing the pH in the presence of phosphate buffer. The 3-O-sulfated oligosaccharide standards are reagents to characterize 3-O-sulfation in HS derived from biological sources.