Rapid Differentiation of Chondroitin Sulfate Isomers by Gas-phase Hydrogen-deuterium Exchange.
Rapid Differentiation of Chondroitin Sulfate Isomers by Gas-phase Hydrogen-deuterium Exchange.
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DOI：
10.2174/1566524020666200915110707
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发表时间：
2020
影响因子：
2.5
通讯作者：
Guttman M
中科院分区：
文献类型：
--
作者：
Alonge KM;Harkewicz R;Guttman M
硫酸软骨素(CS)-糖胺多聚糖(GAG)是附着在CS蛋白多糖上的带负电荷的线性多糖，是中枢和外周组织生物基质的主要成分。据预测，它们与CS二糖结合的硫酸盐基团的位置会影响蛋白质-糖链的相互作用和生物功能。尽管生物组织中CS-GAG的传统免疫组织化学分析提供了发育和疾病状态下GAG丰度的变化信息，但由于分离CS异构体的固有复杂性，对其特定的硫化模式的定量分析受到限制。虽然已经开发了用液相分离来分析和定量硫酸盐异构体的方法，但仍需要新的技术来阐明CS-GAGS的全部生物学特性。在这里，我们研究了离子迁移率光谱和气相氢-重离子交换来解析最常见的硫酸盐化4S-和6S-CS二糖中的位置硫化异构体。这两种异构体的迁移率非常相似，不能用任何漂移气体有效地拆分。相反，气相氢-氚交换显示出与几种氚交换试剂截然不同的吸氢速率，这为分离CS异构体提供了一种新的、快速的方法。气相氢/氚交换是区分硫酸软骨素同分异构体的有效工具。
Chondroitin sulfate (CS)-glycosaminoglycans (GAGs) are linear, negatively charged polysaccharides attached to CS proteoglycans that make up a major component of biological matrices throughout both central and peripheral tissues. The position of their attached sulfate groups to the CS disaccharide is predicted to influence protein-glycan interactions and biological function. Although traditional immunohistochemical analysis of CS-GAGs in biological tissues have provided information regarding changes in GAG abundance during developmental and disease states, quantitative analysis of their specific sulfation patters is limited due to the inherent complexity of separating CS isomers. While methods have been developed to analyze and quantify sulfation isomers using liquid phase separation, new techniques are still needed to elucidate the full biology of CS-GAGs. Here we examine ion mobility spectrometry and gas-phase hydrogen deuterium exchange to resolve positional sulfation isomers in the most common sulfated 4S- and 6S-CS disaccharides. The mobilities for these two isomers are highly similar and could not be resolved effectively with any drift gas tested. In contrast, gas-phase hydrogen deuterium exchange showed very different rates of deuterium uptake with several deuterium exchange reagents, presenting a promising novel and rapid approach for resolving CS isomers. Gas-phase hydrogen/deuterium exchange is an effective tool for distinguishing chondroitin sulfate isomers.