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Genetically encoding Nε-acetyllysine in recombinant proteins

Nature Chemical Biology volume 4, pages 232–234 (2008)Cite this article

Abstract

Nε-acetylation of lysine (1) is a reversible post-translational modification with a regulatory role that rivals that of phosphorylation in eukaryotes. No general methods exist to synthesize proteins containing Nε-acetyllysine (2) at defined sites. Here we demonstrate the site-specific incorporation of Nε-acetyllysine in recombinant proteins produced in Escherichia coli via the evolution of an orthogonal Nε-acetyllysyl-tRNA synthetase/tRNACUA pair. This strategy should find wide applications in defining the cellular role of this modification.

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Figure 1: The MbPylRS/MbtRNACUA pair efficiently and specifically incorporates Cyc in response to an amber stop codon in the gene for myoglobin.
Figure 2: Design and evolution of an MbPylRS/tRNACUA pair for the genetic incorporation of Nε-acetyllysine.

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Acknowledgements

We are grateful to K. Wang (Medical Research Council Laboratory of Molecular Biology, Cambridge) for assisting with early stages of the project, and to P.G. Schultz (Scripps Institute) for materials. J.W.C. is a European Molecular Biology Organization Young Investigator. This work was funded by the Medical Research Council, UK.

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  1. Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 0QH, England, UK

    Heinz Neumann, Sew Y Peak-Chew & Jason W Chin

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  1. Heinz Neumann
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  2. Sew Y Peak-Chew
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  3. Jason W Chin
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Correspondence to Jason W Chin.

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Supplementary Figures 1–4 and Supplementary Methods (PDF 1436 kb)

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Neumann, H., Peak-Chew, S. & Chin, J. Genetically encoding Nε-acetyllysine in recombinant proteins. Nat Chem Biol 4, 232–234 (2008). https://doi.org/10.1038/nchembio.73

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