Kinetic controlled affinity labeling of target enzyme with thioester chemistry
Kinetic controlled affinity labeling of target enzyme with thioester chemistry
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使用硫酯化学对目标酶进行动力学控制的亲和力标记
DOI：
10.1016/j.bmc.2016.05.059
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
S.
中科院分区：
文献类型：
--
作者：
Tomohiro;T.; Nakabayashi;M.; Sugita;Y.; Morimoto;S.
高特异性一直是亲和标记靶标记法的一个重要特征。特别是，通过消耗配体(探针)的快速反应来标记靶标，与靶蛋白的常见官能团反应应具有高度的特异性，而不应与相似的非靶蛋白基团反应。在这里，我们证明了动力学控制的亲和标记酰基辅酶A合成酶使用含有苯硫酯连接的脂肪酸类似物。通过提高结合袋中的标记率，获得了高特异性。该方法可用于分析信号转导途径中的一系列靶标酶和转运蛋白。
High specificity has been an important feature in affinity labeling for target profiling. Especially, to label targets via rapidly progressing reactions with consumption of ligand (probe), high specificity of reaction with common functional groups of target protein should be achieved without reactions with similar groups of non-target proteins. Herein, we demonstrate the kinetic controlled affinity labeling of acyl CoA synthetase using a fatty acid analogue containing a phenylthioester linkage. High specificity was attained by accelerating the labeling rate in the binding pocket. This approach could be useful for profiling a series of target enzymes and transporters in signal transduction pathways.