Tunable light and drug induced depletion of target proteins
Tunable light and drug induced depletion of target proteins
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DOI：
10.1038/s41467-019-14160-8
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发表时间：
2020-01-16
影响因子：
16.6
通讯作者：
Leonhardt, Heinrich
中科院分区：
文献类型：
--
作者：
Deng, Wen;Bates, Jack A.;Leonhardt, Heinrich
发育和疾病中的生物过程由细胞蛋白质的丰度、定位和修饰控制。我们已经开发了基于重组E3泛素连接酶的通用工具，所述重组E3泛素连接酶由光或药物诱导的异源二聚化控制，用于细胞和生物体中内源性蛋白质的纳米抗体或DARPin靶向消耗。我们使用这种快速的，可调的和可逆的蛋白质耗竭的功能研究的基本蛋白质，如PCNA在DNA修复和研究CED-3在秀丽隐杆线虫发育过程中的细胞凋亡的作用。这些独立的工具可以组合用于不同蛋白质组的空间和时间消耗，可以帮助区分即时细胞反应和长期适应效应，并可以促进复杂网络的探索。
Biological processes in development and disease are controlled by the abundance, localization and modification of cellular proteins. We have developed versatile tools based on recombinant E3 ubiquitin ligases that are controlled by light or drug induced heterodimerization for nanobody or DARPin targeted depletion of endogenous proteins in cells and organisms. We use this rapid, tunable and reversible protein depletion for functional studies of essential proteins like PCNA in DNA repair and to investigate the role of CED-3 in apoptosis during Caenorhabditis elegans development. These independent tools can be combined for spatial and temporal depletion of different sets of proteins, can help to distinguish immediate cellular responses from long-term adaptation effects and can facilitate the exploration of complex networks.