Synthetic and Functional Biomolecules Center, Beijing National Laboratory for Molecular Sciences, Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education, College of Chemistry and Molecular Engineering, Peking University, Beijing 100871, China.
Peking-Tsinghua Center for Life Sciences, Beijing 100871, China.
Nat Chem. 2014 Apr;6(4):352-61. doi: 10.1038/nchem.1887. Epub 2014 Mar 16.
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.
利用小分子或化学试剂来调节细胞内蛋白质的功能，特别是以获得功能的方式，仍然是一个挑战。与基于抑制剂的失活方法相比，基于功能获得的方法能够在细胞内激活特定的信号通路。在这里，我们报告了一种使用钯介导的去保护反应在活细胞内激活蛋白质的化学拯救策略。我们鉴定出了生物相容性和高效的钯催化剂，它们可以切割保护的赖氨酸类似物的炔丙基碳酸酯基团，生成游离的赖氨酸。赖氨酸类似物可以被基因和位点特异性地掺入蛋白质中，从而可以控制反应位点。该去保护策略适用于一系列不同的细胞系和蛋白质。我们进一步将这种生物相容性的保护基/催化剂对用于细菌 III 型效应蛋白中关键赖氨酸残基的笼状和随后的释放，这揭示了其毒力机制的细节。
