Study of Highly Selective and Efficient Thiol Derivatization Using Selenium Reagents by Mass Spectrometry
Study of Highly Selective and Efficient Thiol Derivatization Using Selenium Reagents by Mass Spectrometry
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硒试剂质谱高选择性高效硫醇衍生化研究
DOI：
10.1021/ac1011602
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发表时间：
2010-08-15
影响因子：
7.4
通讯作者：
Chen, Hao
中科院分区：
文献类型：
--
作者：
Xu, Kehua;Zhang, Yun;Chen, Hao
本文报道了一种标记生物硫醇的新策略的系统的质谱学研究，包括硫醇裂解Se-N键形成新的Se-S键。我们的数据表明，该反应具有高选择性、快速、可逆和高效的特点。在20个氨基酸中，只有半胱氨酸对含Se-N的试剂有反应，反应在几秒钟内发生。通过添加二硫苏糖醇，可以回收硒试剂衍生的多肽。高的反应选择性和可逆性为从混合物中选择性识别和分离硫醇提供了潜力。此外，根据所使用的硒试剂的不同，衍生的多肽离子表现出可调的解离行为(要么容易断裂，要么在碰撞诱导解离时保持形成的Se-S键)，这一特征在蛋白质组学研究中是有用的。同样重要的是，硫醇衍生化的产率是惊人的，在30 S内，Ebselen对蛋白质β-乳球蛋白A的转化率为100%。此外，还展示了从混合物中快速筛选硫醇肽和鉴定游离和结合硫醇的数量的初步应用。这项研究中发现的独特的硒化学对于蛋白质/多肽的硫醇和二硫键的MS分析将是有价值的。
This paper reports a systemic mass spectrometry (MS) investigation of a novel strategy for labeling biological thiols, involving the cleavage of the Se-N bond by thiol to form a new Se-S bond. Our data show that the reaction is highly selective, rapid, reversible, and efficient. Among 20 amino acids, only cysteine is reactive toward Se-N containing reagents and the reaction occurs in seconds. With the addition of dithiothreitol, peptides derivatized by selenium reagents can be recovered. The high reaction selectivity and reversibility provide potential in both selective identification and isolation of thiols from mixtures. Also, with dependence on the selenium reagent used, derivatized peptide ions exhibit tunable dissociation behaviors (either facile cleavage or preservation of the formed Se-S bond upon collision-induced dissociation), a feature that is useful in proteomics studies. Equally importantly, the thiol derivatization yield is striking, as reflected by 100% conversion of protein beta-lactoglobulin A using ebselen within 30 s. In addition, preliminary applications such as rapid screening of thiol peptides from mixtures and identification of the number of protein free and bound thiols have been demonstrated. The unique selenium chemistry uncovered in this study would be valuable in the MS analysis of thiols and disulfide bonds of proteins/peptides.