Vendrell-Navarro Glòria, Brockmeyer Andreas, Waldmann Herbert, Janning Petra, Ziegler Slava
Abteilung Chemische Biologie, Max-Planck-Institut für molekulare Physiologie, Otto-Hahn-Str. 11, Dortmund, 44227, Germany.
Methods Mol Biol. 2015;1263:263-86. doi: 10.1007/978-1-4939-2269-7_21.
Currently, cell-based screenings yield a multitude of small molecule modulators of diverse biological processes. The most demanding step in the course of elucidation of the mode of action of biologically active compounds is the identification of the target proteins. Although there is no generic approach available, affinity-based chemical proteomics is the most widely applied methodology. Particularly, quantitative chemical proteomics has proven very powerful in the identification of the putative targets of small molecules. Here we describe the procedure for identification of target proteins for small molecules employing affinity chromatography and the stable isotope labeling in cell culture (SILAC) for quantitative proteomics.
目前，基于细胞的筛选产生了众多调节各种生物过程的小分子调节剂。在阐明生物活性化合物作用方式的过程中，最具挑战性的步骤是鉴定靶蛋白。虽然没有通用的方法，但基于亲和力的化学蛋白质组学是应用最广泛的方法。特别是，定量化学蛋白质组学已被证明在鉴定小分子的假定靶标方面非常有效。在这里，我们描述了使用亲和色谱和细胞培养中的稳定同位素标记（SILAC）进行定量蛋白质组学来鉴定小分子靶蛋白的程序。
