N-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 N-acetyllysine (2) at defined sites. Here we demonstrate the site-specific incorporation of N-acetyllysine in recombinant proteins produced in Escherichia coli via the evolution of an orthogonal N-acetyllysyl-tRNA synthetase/tRNA_CUA pair. This strategy should find wide applications in defining the cellular role of this modification.

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

Correspondence to: Jason W Chin¹ e-mail: chin@mrc-lmb.cam.ac.uk

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