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Palladium-triggered deprotection chemistry for protein activation in living cells

Nature Chemistry volume 6, pages 352–361 (2014)Cite this article

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Abstract

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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Figure 1: A chemical decaging strategy for protein activation in living cells.
Figure 2: Catalyst screening for palladium-mediated deprotection reactions.
Figure 3: Identification of biocompatible and efficient palladium reagents for propargyl carbamate cleavage on intact proteins.
Figure 4: Palladium-mediated propargyl carbamate cleavage within living cells.
Figure 5: Modulating the activity of a bacterial phosphothreonine lyase OspF via palladium-mediated decaging of lysine in vitro and in vivo.
Figure 6: Chemically rescued OspF as a tool to study the intracellular localization of its cognitive substrate Erk.

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Acknowledgements

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.

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Authors and Affiliations

  1. 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

    Jie Li, Jingyi Zhao, Siqi Zheng, Shixian Lin, Long Chen, Maiyun Yang, Shang Jia & Peng R. Chen

  2. Peking-Tsinghua Center for Life Sciences, Beijing 100871, China

    Juntao Yu, Jie Wang & Peng R. Chen

  3. College of Chemistry and Chemical Engineering, Lanzhou University, Lanzhou, 730000, China

    Xiaoyu Zhang

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Contributions

P.R.C. conceived and designed the experiments. J.L., J.Y., J.Z., J.W., S.Z., S.L., L.C., M.Y., S.J. and X.Z. performed the experiments. P.R.C., J.L., J.Y., J.Z. and J.W. analysed the data. P.R.C, J.L. and J.Y. prepared the figures and co-wrote the paper, with input from all the authors.

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Correspondence to Peng R. Chen.

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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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