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.
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This work was supported by research grants from National Natural Science Foundation of China (21225206 and 91313301) and the National Key Basic Research Foundation of China (2010CB912302). We thank F. Shao and O. Schneewind for the donation of plasmids.
The authors declare no competing financial interests.
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Li, J., Yu, J., Zhao, J. et al. Palladium-triggered deprotection chemistry for protein activation in living cells. Nature Chem 6, 352–361 (2014). https://doi.org/10.1038/nchem.1887
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