Palladium-triggered deprotection chemistry for protein activation in living cells
Palladium-triggered deprotection chemistry for protein activation in living cells
复制标题
DOI：
10.1038/nchem.1887
复制
发表时间：
2014-04-01
期刊：
影响因子：
21.8
通讯作者：
Chen, Peng R.
中科院分区：
文献类型：
--
作者：
Li, Jie;Yu, Juntao;Chen, Peng R.
采用小分子或化学试剂来调节细胞内蛋白质的功能，特别是以功能获得的方式，仍然是一个挑战。与基于通道的功能丧失方法相比，基于功能获得的方法能够在细胞内激活特定的信号传导途径。在这里，我们报告了一种化学救援策略，使用钯介导的脱保护反应，激活活细胞内的蛋白质。我们确定了生物相容性和有效的钯催化剂，裂解炔丙基氨基甲酸酯基团的保护赖氨酸类似物，以产生游离赖氨酸。赖氨酸类似物可以遗传地和位点特异性地掺入蛋白质中，这使得能够控制反应位点。这种脱保护策略显示出对一系列不同的细胞系和蛋白质起作用。我们进一步将这种生物相容性保护基团/催化剂对用于在宿主细胞内笼化和随后释放细菌III型效应蛋白中的关键赖氨酸残基，这揭示了其毒力机制的细节。
Employing small molecules or chemical reagents to modulate the function of an intracellular protein, particularly in a gain-of-function fashion, remains a challenge. In contrast to inhibitor-based loss-of-function approaches, methods based on a gain of function enable specific signalling pathways to be activated inside a cell. Here we report a chemical rescue strategy that uses a palladium-mediated deprotection reaction to activate a protein within living cells. We identify biocompatible and efficient palladium catalysts that cleave the propargyl carbamate group of a protected lysine analogue to generate a free lysine. The lysine analogue can be genetically and site-specifically incorporated into a protein, which enables control over the reaction site. This deprotection strategy is shown to work with a range of different cell lines and proteins. We further applied this biocompatible protection group/catalyst pair for caging and subsequent release of a crucial lysine residue in a bacterial Type III effector protein within host cells, which reveals details of its virulence mechanism.