Structurally Informative Tandem Mass Spectrometry of Highly Sulfated Natural and Chemoenzymatically Synthesized Heparin and Heparan Sulfate Glycosaminoglycans
Structurally Informative Tandem Mass Spectrometry of Highly Sulfated Natural and Chemoenzymatically Synthesized Heparin and Heparan Sulfate Glycosaminoglycans
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DOI：
10.1074/mcp.m112.026880
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发表时间：
2013-04-01
影响因子：
7
通讯作者：
Amster, I. Jonathan
中科院分区：
文献类型：
--
作者：
Kailemia, Muchena J.;Li, Lingyun;Amster, I. Jonathan
来源于肝素和硫酸乙酰肝素的高度硫酸化的糖胺聚糖寡糖一直是通过串联质谱法分析的高度难处理的一类分子。在许多离子活化方法下，这类分子通常表现出SO 3损失作为最重要的断裂途径，干扰糖胺聚糖链中磺基位置的分配。我们在这里报告的方法，稳定磺基，有利于完整的结构分析密集硫酸化（两个或更多个磺基每二糖重复单元）肝素和硫酸乙酰肝素低聚物。这是通过完全去除所有可电离的质子来实现的，通过在电喷雾电离期间充电或通过Na+/H+交换。向样品溶液中加入毫摩尔水平的NaOH有助于产生符合此标准的前体离子。发现这种方法适用于多种来源于天然来源或通过化学酶合成产生的硫酸肝素寡糖，其具有多达12个糖亚基和多达11个磺基。Molecular & Cellular Proteomics 12：10.1074/mcp. M112.026880，979-990，2013.
The highly sulfated glycosaminoglycan oligosaccharides derived from heparin and heparan sulfate have been a highly intractable class of molecules to analyze by tandem mass spectrometry. Under the many methods of ion activation, this class of molecules generally exhibits SO3 loss as the most significant fragmentation pathway, interfering with the assignment of the location of sulfo groups in glycosaminoglycan chains. We report here a method that stabilizes sulfo groups and facilitates the complete structural analysis of densely sulfated (two or more sulfo groups per disaccharide repeat unit) heparin and heparan sulfate oligomers. This is achieved by complete removal of all ionizable protons, either by charging during electrospray ionization or by Na+/H+ exchange. The addition of millimolar levels of NaOH to the sample solution facilitates the production of precursor ions that meet this criterion. This approach is found to work for a variety of heparin sulfate oligosaccharides derived from natural sources or produced by chemoenzymatic synthesis, with up to 12 saccharide subunits and up to 11 sulfo groups. Molecular & Cellular Proteomics 12: 10.1074/mcp.M112.026880, 979-990, 2013.