Mass spectrometric method for determining the uronic acid epimerization in heparan sulfate disaccharides generated using nitrous acid.
Mass spectrometric method for determining the uronic acid epimerization in heparan sulfate disaccharides generated using nitrous acid.
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DOI：
10.1021/ac3016054
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发表时间：
2012-09-04
影响因子：
7.4
通讯作者：
Zaia, Joseph
中科院分区：
文献类型：
--
作者：
Gill, Vanessa Leah;Wang, Qi;Shi, Xiaofeng;Zaia, Joseph
硫酸乙酰肝素（HS）糖胺聚糖（GAG）调节许多生物学功能。为了更好地了解它们的生物学作用，有必要了解HS的结构，这需要鉴定硫酸化模式以及糖醛酸差向异构化。为了模拟HS结构，有必要对解聚产物进行定量分析。迄今为止，已经显示了用于分析肝素裂解酶分解产物的液相色谱-质谱（LC-MS）方法。然而，这些酶破坏了糖醛酸差向异构化的信息。使用亚硝酸（HONO）的脱氨基裂解是保留糖醛酸差向异构化的GAG解聚的经典方法。几种色谱方法已用于分析脱氨基裂解产物。色谱法的缺点是不能直接读出产生观察到的峰的结构。这份报告演示了多孔石墨化碳（PGC）-MS方法的量化HONO生成的二糖，以获得有关的硫酸化模式和糖醛酸差向异构化的信息。在这里，我们演示了糖醛酸差向异构体以及几何硫酸化异构体的分离和鉴定。结果与基准HS和肝素样品的预期结果相当。这些数据证明了PGC-MS用于定量HS亚硝酸解聚产物以用于HS和肝素的结构分析的实用性。
Heparan sulfate (HS) glycosaminoglycans (GAGs) regulate a host of biological functions. To better understand their biological roles, it is necessary to gain understanding about the structure of HS, which requires identification of the sulfation pattern as well as the uronic acid epimerization. In order to model HS structure, it is necessary to quantitatively profile depolymerization products. To date, liquid chromatography-mass spectrometry (LC-MS) methods for profiling heparin lyase decomposition products have been shown. These enzymes, however, destroy information about uronic acid epimerization. Deaminative cleavage using nitrous acid (HONO) is a classic method for GAG depolymerization that retains uronic acid epimerization. Several chromatographic methods have been used for analysis of deaminative cleavage products. The chromatographic methods have the disadvantage that there is no direct readout on the structures producing the observed peaks. This report demonstrates a porous graphitized carbon (PGC)-MS method for the quantification of HONO generated disaccharides to obtain information about the sulfation pattern and uronic acid epimerization. Here we demonstrate the separation and identification of uronic acid epimers as well as geometric sulfation isomers. The results are comparable to those expected for benchmark HS and heparin samples. The data demonstrate the utility of PGC-MS for quantification of HS nitrous acid depolymerization products for structural analysis of HS and heparin.
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影响因子：
4.8
作者：
Maccarana, M;Sakura, Y;Lindahl, U
通讯作者：
Lindahl, U
影响因子：
4.3
作者：
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影响因子：
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作者：
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DOI：
10.1016/j.jchromb.2005.07.014
发表时间：
2005-09-25
影响因子：
3
作者：
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通讯作者：
Whitelock, JM
影响因子：
4.8
作者：
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通讯作者：
Dierks, Thomas