Enzymatic/Chemical Release of O-Glycans Allowing MS Analysis at High Sensitivity
Enzymatic/Chemical Release of O-Glycans Allowing MS Analysis at High Sensitivity
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DOI：
10.1021/ac901363h
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发表时间：
2009-12-01
影响因子：
7.4
通讯作者：
Mechref, Yehia
中科院分区：
文献类型：
--
作者：
Goetz, John A.;Novotny, Milos V.;Mechref, Yehia
随着O-连接寡糖在许多医学疾病中的作用越来越重要，必须开发新技术，以高灵敏度进行分析。虽然质谱法（MS）提供了足够的测量重要的O-连接寡糖聚糖和他们的档案，从糖蛋白的释放尚未得到充分解决的生物医学应用的需要。这项工作描述了一种新的策略，涉及一个完整的酶促降解与O-连接的寡糖的固相全甲基化过程中的化学释放相结合。分析数据表明，在全甲基化过程中，丝氨酸和苏氨酸（而不是精氨酸）残基的切割非常有效。串联MS分析证实了模型糖蛋白的这些观察结果。通过同位素标记MS的比较测量表明，这种方法大大优于以前使用的裂解程序上级。
As the role of O-linked oligosaccharides have been demonstrated to be increasingly important in numerous medical conditions, it is imperative to develop new techniques allowing their analysis at high sensitivity. While mass spectrometry (MS) provides adequate measurements of important O-linked oligosaccharides glycans and their profiles, the release from glycoproteins has not been sufficiently addressed for the needs of biomedical applications. This work describes a new strategy, involving the combination of a complete enzymatic degradation with a chemical release during the solid-phase permethylation of O-linked oligosaccharides. The analytical data implicate highly effective cleavage from the serine and threonine (but not arginine) residues, during permethylation. Tandem MS analyses confirmed these observations for model glycoproteins. Comparative measurements through isotopic labeling MS show this approach to be vastly superior over the previously used cleavage procedures.