A Simple and Effective Cleavable Linker for Chemical Proteomics Applications
A Simple and Effective Cleavable Linker for Chemical Proteomics Applications
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DOI：
10.1074/mcp.m112.021014
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发表时间：
2013-01-01
影响因子：
7
通讯作者：
Verhelst, Steven H. L.
中科院分区：
文献类型：
--
作者：
Yang, Yinliang;Hahne, Hannes;Verhelst, Steven H. L.
代谢性标记或探针修饰蛋白质的研究是化学蛋白质组学的一个重要领域。蛋白质靶标的分离和纯化是MS鉴定前的必要步骤。生物素-链霉亲和素系统在这一过程中得到了广泛的应用，但苛刻的变性条件也会释放天然的生物素化蛋白和非选择性结合蛋白。可切割链接器策略是解决这一问题的一种很有前途的方法。虽然已经开发和测试了几个可切割接头，但一种高效、容易合成和廉价的可切割接头是蛋白质组学工具箱的理想补充。在这里，我们描述了一种邻近的二醇可裂解连接子的化学蛋白质组学应用。通过易于处理的化学，我们将这种连接物结合到一个基于活性的探针和一个生物素炔标签中，使其能够进行生物正交连接。有了这些试剂，与标准的珠状消化相比，背景蛋白鉴定显著减少。分子与细胞蛋白质组学12：10.1074/mcp.M112.021014,237-244,2013年。
The study of metabolically labeled or probe-modified proteins is an important area in chemical proteomics. Isolation and purification of the protein targets is a necessary step before MS identification. The biotin-streptavidin system is widely used in this process, but the harsh denaturing conditions also release natively biotinylated proteins and non-selectively bound proteins. A cleavable linker strategy is a promising approach for solving this problem. Though several cleavable linkers have been developed and tested, an efficient, easily synthesized, and inexpensive cleavable linker is a desirable addition to the proteomics toolbox. Here, we describe the chemical proteomics application of a vicinal diol cleavable linker. Through easy-to-handle chemistry we incorporate this linker into an activity-based probe and a biotin alkyne tag amenable for bioorthogonal ligation. With these reagents, background protein identifications are significantly reduced relative to standard on-bead digestion. Molecular & Cellular Proteomics 12: 10.1074/mcp.M112.021014, 237-244, 2013.