Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins.
Thiol-ene-Enabled Detection of Thiophosphorylation as a Labeling Strategy for Phosphoproteins.
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硫醇烯检测硫代磷酸化作为磷蛋白的标记策略。
DOI：
10.1007/978-1-4939-3049-4_1
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Carlson,ErinE
中科院分区：
文献类型：
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作者：
Wilke,KaelynE;Carlson,ErinE
三磷酸腺苷 (ATP) 类似物腺苷 5′-O-(3-硫代三磷酸) (ATPγS) 已被用作研究激酶底物磷酸化的工具。硫代磷酸基团的转移不仅代表了磷酸蛋白质组中的独特修饰，而且对磷酸酶活性也保持稳定。然而，由于硫代磷酸盐和蛋白质硫醇具有相似的化学反应性，该物种的检测很复杂。在这里，我们描述了一种利用硫醇-烯反应检测蛋白质硫代磷酸化的新方法。首先通过自由基化学化学选择性地封端蛋白质硫醇，可以特异性地观察激酶催化的硫代磷酸化。
The adenosine triphosphate (ATP) analogue adenosine 5′-O-(3-thiotriphosphate) (ATPγS) has been applied as a tool to study kinase-substrate phosphorylation. Not only does the transfer of a thiophosphate group represent a unique modification amid the phosphoproteome, but it can also be stable to phosphatase activity. However, detection of this species is complicated due to the similar chemical reactivity of thiophosphate and proteinaceous thiols. Here, we describe a novel method for detection of protein thiophosphorylation utilizing the thiol-ene reaction. By first chemoselectively capping protein thiols through radical chemistry, kinase-catalyzed thiophosphorylation can be visualized specifically.