Proteome-Wide Profiling of Targets of Cysteine reactive Small Molecules by Using Ethynyl Benziodoxolone Reagents
Proteome-Wide Profiling of Targets of Cysteine reactive Small Molecules by Using Ethynyl Benziodoxolone Reagents
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DOI：
10.1002/anie.201505641
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发表时间：
2015-09-07
影响因子：
16.6
通讯作者：
Adibekian, Alexander
中科院分区：
文献类型：
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作者：
Abegg, Daniel;Frei, Reto;Adibekian, Alexander
在这项研究中，我们提出了一个高效的方法，在复杂的蛋白质组和活细胞中的半胱氨酸残基的蛋白质组学分析。我们的方法是基于使用带有叠氮基的可点击的炔基苯碘多索酮的复杂蛋白质组中的半胱氨酸的炔基化。该反应在温和的生理条件下快速进行，并且具有非常高的化学选择性。所形成的叠氮化物封端的炔基-半胱氨酸加合物易于通过LC-MS/MS检测，并且可以通过CuAAC用塔姆拉或生物素炔进一步官能化。我们通过鉴定姜黄素的蛋白质组学靶点，证明了炔基苯并碘多佐酮在化学蛋白质组学应用中的实用性，姜黄素是一种二芳基庚烷类天然产物，过去和现在都是作为抗癌剂的多项人体临床试验的一部分。我们的研究结果表明，姜黄素共价修饰细胞信号传导和代谢的几个关键参与者，最引人注目的是酪蛋白激酶I γ。我们预计，这种新的半胱氨酸分析方法将在化学蛋白质组学和药物发现中找到广泛的应用。
In this study, we present a highly efficient method for proteomic profiling of cysteine residues in complex proteomes and in living cells. Our method is based on alkynylation of cysteines in complex proteomes using a clickable alkynyl benziodoxolone bearing an azide group. This reaction proceeds fast, under mild physiological conditions, and with a very high degree of chemoselectivity. The formed azide-capped alkynyl-cysteine adducts are readily detectable by LC-MS/MS, and can be further functionalized with TAMRA or biotin alkyne via CuAAC. We demonstrate the utility of alkynyl benziodoxolones for chemical proteomics applications by identifying the proteomic targets of curcumin, a diarylheptanoid natural product that was and still is part of multiple human clinical trials as anticancer agent. Our results demonstrate that curcumin covalently modifies several key players of cellular signaling and metabolism, most notably the enzyme casein kinase I gamma. We anticipate that this new method for cysteine profiling will find broad application in chemical proteomics and drug discovery.