Chemical Development of Intracellular Protein Heterodimerizers
Chemical Development of Intracellular Protein Heterodimerizers
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DOI：
10.1016/j.chembiol.2013.03.010
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发表时间：
2013-04-18
影响因子：
--
通讯作者：
Wymann, Matthias P.
中科院分区：
文献类型：
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作者：
Erhart, Dominik;Zimmermann, Mirjam;Wymann, Matthias P.
由受体配体或癌基因启动的细胞激活触发复杂和复杂的细胞内信号传导。在确定的起点和细胞位置启动信号的技术对于阐明选定途径的输出是有吸引力的。在这里，我们提出了基于源自Halo标签和snap标签的小分子交联融合蛋白的蛋白质异源二聚化系统的开发和验证。Halo标签和snap标签（HaXS）的化学二聚体具有良好的选择性，并对细胞内反应性进行了优化。HaXS强迫蛋白质相互作用，并能将蛋白质转运到不同的细胞室。由于HaloTag-HaXS-SNAP-tag复合物的共价性质，可以很容易地监测细胞内二聚化。最初的应用包括蛋白靶向细胞骨架、质膜、溶酶体，启动PI3K/mTOR通路，以及与雷帕霉素二聚化系统结合形成多重蛋白复合物。
Cell activation initiated by receptor ligands or oncogenes triggers complex and convoluted intracellular signaling. Techniques initiating signals at defined starting points and cellular locations are attractive to elucidate the output of selected pathways. Here, we present the development and validation of a protein heterodimerization system based on small molecules cross-linking fusion proteins derived from Halo Tags and SNAP-tags. Chemical dimerizers of Halo Tag and SNAP-tag (HaXS) show excellent selectivity and have been optimized for intracellular reactivity. HaXS force protein-protein interactions and can translocate proteins to various cellular compartments. Due to the covalent nature of the HaloTag-HaXS-SNAP-tag complex, intracellular dimerization can be easily monitored. First applications include protein targeting to cytoskeleton, to the plasma membrane, to lysosomes, the initiation of the PI3K/mTOR pathway, and multiplexed protein complex formation in combination with the rapamycin dimerization system.