Degradation of HaloTag-fused nuclear proteins using bestatin-HaloTag ligand hybrid molecules
Degradation of HaloTag-fused nuclear proteins using bestatin-HaloTag ligand hybrid molecules
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DOI：
10.1039/c5ob01395j
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发表时间：
2015-01-01
影响因子：
3.2
通讯作者：
Ishikawa, Minoru
中科院分区：
文献类型：
--
作者：
Tomoshige, Shusuke;Naito, Mikihiko;Ishikawa, Minoru
我们已经开发了一种蛋白质敲低技术，使用杂交小分子设计为靶蛋白配体和泛素连接酶细胞凋亡蛋白1 （cIAP1）配体的偶联物。然而，这项技术有一些局限性。在这里，我们报告了一种新的蛋白质敲低系统的发展，以解决这些限制。在这个系统中，目标蛋白与HaloTag融合，为降解诱导剂提供一个共同的结合位点。我们设计并合成了由烷基氯组成的小分子作为HaloTag结合降解诱导剂，它与HaloTag结合，与BE04(2)连接，BE04与cIAP1结合。利用该系统，我们成功敲除了活细胞中与配体未知的核蛋白HaloTag-fused cAMP响应元件结合蛋白1 （HaloTag-CREB1）和HaloTag-fused c-jun （HaloTag-c-jun）。halotag结合降解诱导剂易于合成，有望成为泛降解halotag融合蛋白的生物工具。
We have developed a protein knockdown technology using hybrid small molecules designed as conjugates of a ligand for the target protein and a ligand for ubiquitin ligase cellular inhibitor of apoptosis protein 1 (cIAP1). However, this technology has several limitations. Here, we report the development of a novel protein knockdown system to address these limitations. In this system, target proteins are fused with HaloTag to provide a common binding site for a degradation inducer. We designed and synthesized small molecules consisting of alkyl chloride as the HaloTag-binding degradation inducer, which binds to HaloTag, linked to BE04 (2), which binds to cIAP1. Using this system, we successfully knocked down HaloTag-fused cAMP responsive element binding protein 1 (HaloTag-CREB1) and HaloTag-fused c-jun (HaloTag-c-jun), which are ligand-unknown nuclear proteins, in living cells. HaloTag-binding degradation inducers can be synthesized easily, and are expected to be useful as biological tools for pan-degradation of HaloTag-fused proteins.