Targeted Protein Degradation through Fast Optogenetic Activation and Its Application to the Control of Cell Signaling
Targeted Protein Degradation through Fast Optogenetic Activation and Its Application to the Control of Cell Signaling
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DOI：
10.1021/jacs.1c04324
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发表时间：
2021-06-14
影响因子：
15
通讯作者：
Deiters, Alexander
中科院分区：
文献类型：
--
作者：
Ryan, Amy;Liu, Jihe;Deiters, Alexander
光学触发的蛋白质降解方法的发展使得能够研究动态蛋白质功能，例如参与细胞信号传导的那些功能，这些功能难以用传统的遗传技术来探测。在这里，我们描述了一种新的光控肽降解决定子的设计和实施赋予N端途径降解其蛋白质的目标。降解决定子包含光笼化的N-末端氨基酸和富含赖氨酸的13-残基接头。通过笼N-末端残基，我们能够光学控制N-降解决定子识别的E3连接酶，从而控制泛素化和蛋白酶体降解的目标蛋白。我们通过将这种方法应用于一组不同的靶蛋白，包括EGFP，萤火虫荧光素酶，激酶MEK 1和磷酸酶DUSP 6（也称为MKP 3），证明了广泛的适用性。笼状降解决定子可以与最小的蛋白质工程一起使用，并在秒至分钟的时间尺度上提供几乎完全的光触发蛋白质降解。
Development of methodologies for optically triggered protein degradation enables the study of dynamic protein functions, such as those involved in cell signaling, that are difficult to be probed with traditional genetic techniques. Here, we describe the design and implementation of a novel light-controlled peptide degron conferring N-end pathway degradation to its protein target. The degron comprises a photocaged N-terminal amino acid and a lysine-rich, 13-residue linker. By caging the N-terminal residue, we were able to optically control N-degron recognition by an E3 ligase, consequently controlling ubiquitination and proteasomal degradation of the target protein. We demonstrate broad applicability by applying this approach to a diverse set of target proteins, including EGFP, firefly luciferase, the kinase MEK1, and the phosphatase DUSP6 (also known as MKP3). The caged degron can be used with minimal protein engineering and provides virtually complete, light-triggered protein degradation on a second to minute time scale.