The chemoselective one-step alkylation and isolation of thiophosphorylated cdk2 substrates in the presence of native cysteine.
The chemoselective one-step alkylation and isolation of thiophosphorylated cdk2 substrates in the presence of native cysteine.
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在天然半胱氨酸存在下，硫代磷酸化 cdk2 底物的化学选择性一步烷基化和分离。
DOI：
10.1002/cbic.201000528
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发表时间：
2011
期刊：
影响因子：
--
通讯作者：
Lee SE
中科院分区：
文献类型：
--
作者：
Lee SE
信号通路的阐明在很大程度上依赖于蛋白激酶底物的鉴定。最近的研究已经证明了使用ATP类似物和修饰的蛋白激酶进行特异性底物标记的化学遗传学的功效。在这里，我们将N6-（环己基）ATPγS与类似物敏感的cdk 2变体结合，使其底物硫代磷酸化，并证明了pH依赖性，化学选择性，一步烷基化，以促进硫代磷酸化肽的检测或分离。
The elucidation of signalling pathways relies heavily upon the identification of protein kinase substrates. Recent investigations have demonstrated the efficacy of chemical genetics using ATP analogues and modified protein kinases for specific substrate labelling. Here we combineN6‐(cyclohexyl)ATPγS with an analogue‐sensitive cdk2 variant to thiophosphorylate its substrates and demonstrate a pH‐dependent, chemoselective, one‐step alkylation to facilitate the detection or isolation of thiophosphorylated peptides.