Compositional profiling of heparin/heparan sulfate using mass spectrometry: assay for specificity of a novel extracellular human endosulfatase
Compositional profiling of heparin/heparan sulfate using mass spectrometry: assay for specificity of a novel extracellular human endosulfatase
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DOI：
10.1093/glycob/cwi064
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发表时间：
2005-08-01
期刊：
影响因子：
4.3
通讯作者：
Leary, JA
中科院分区：
文献类型：
--
作者：
Saad, OM;Ebel, H;Leary, JA
肝素和硫酸肝素（HS）是糖生物学领域研究的一类重要碳水化合物，具有多种生物学功能。它们的硫酸化模式和结构域组织的变化与不同的病理情况有关，如病毒感染、肿瘤生长和转移。为了获得这些生物分子的结构信息，以及它们在细胞生长和发育的不同阶段可能经历的修饰，研究人员开发了一种基于质谱的方法，并用于获得包含肝素/HS的糖胺聚糖（GAGs）的明确结构信息。该方法被用于检测新发现的人细胞外硫酸酯内切酶HSulf-2的肝素底物特异性，该酶与肿瘤发生有关。这个新方案包含12种已知的肝素双糖，包括三组异构体。为每个双糖确定一个独特的响应因子(R)，而多路复用和数据处理方法被纳入更快的数据采集和量化目的。用从牛和猪组织中分离的各种肝素/HS样品进行了原理验证。
An important class of carbohydrates studied within the field of glycobiology, heparin and heparan sulfate (HS) have been implicated in a diverse array of biological functions. Changes in their sulfation pattern and domain organization have been associated with different pathological situations such as viral infectivity, tumor growth, and metastasis. To obtain structural information about these biomolecules, and the modifications they may undergo during different stages of cell growth and development, a mass spectrometry-based method was developed and used to obtain unambiguous structural information on the glycosaminoglycans (GAGs) that comprise heparin/HS. The method was applied to assay for the heparin substrate specificity of a newly discovered human extracellular endosulfatase, HSulf-2, which has been implicated in tumorigenesis. This new protocol incorporates 12 known heparin disaccharides, including three sets of isomers. A unique response factor (R) is determined for each disaccharide, whereas a multiplexed and data processing method is incorporated for faster data acquisition and quantification purposes. Proof of principle was performed by using various heparin/HS samples isolated from bovine and porcine tissues.