Electrophilic PROTACs that degrade nuclear proteins by engaging DCAF16
Electrophilic PROTACs that degrade nuclear proteins by engaging DCAF16
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DOI：
10.1038/s41589-019-0279-5
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发表时间：
2019-07-01
影响因子：
14.8
通讯作者：
Cravatt, Benjamin F.
中科院分区：
文献类型：
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作者：
Zhang, Xiaoyu;Crowley, Vincent M.;Cravatt, Benjamin F.
配体依赖性蛋白质降解已经成为一种引人注目的策略，以控制细胞的蛋白质含量。然而，到目前为止，只有有限数量的E3连接酶被发现支持这一过程。在这里，我们使用了一种化学蛋白质组学策略，该策略利用广泛反应性的半胱氨酸定向亲电片段与细胞内蛋白质的选择性配体（例如，FKBP 12的SLF，BRD 4的JQ 1）偶联，以筛选通过E3连接酶共价加合作用的异双功能降解剂化合物（或蛋白水解靶向嵌合体，PROTAC）。该方法鉴定了DCAF 16-CUL 4-DDB 1 E3泛素连接酶的一种表征较差的底物识别组分-作为亲电PROTAC的靶标，该亲电PROTAC促进蛋白质的核限制性降解。我们发现，仅需要修饰适度部分（~ 10 - 40%）的DCAF 16以支持蛋白质降解，这表明亲电子PROTAC诱导新底物降解而基本上不干扰参与的E3连接酶的功能的潜力。
Ligand-dependent protein degradation has emerged as a compelling strategy to pharmacologically control the protein content of cells. So far, however, only a limited number of E3 ligases have been found to support this process. Here, we use a chemical proteomic strategy that leverages broadly reactive, cysteine-directed electrophilic fragments coupled to selective ligands for intracellular proteins (for example, SLF for FKBP12, JQ1 for BRD4) to screen for heterobifunctional degrader compounds (or proteolysis targeting chimeras, PROTACs) that operate by covalent adduction of E3 ligases. This approach identified DCAF16-a poorly characterized substrate recognition component of CUL4-DDB1 E3 ubiquitin ligases-as a target of electrophilic PROTACs that promote the nuclear-restricted degradation of proteins. We find that only a modest fraction (-10-40%) of DCAF16 needs to be modified to support protein degradation, pointing to the potential for electrophilic PROTACs to induce neosubstrate degradation without substantially perturbing the function of the participating E3 ligase.