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American Chemical Society, Journal of the American Chemical Society, 28(133), p. 10708-10711, 2011

DOI: 10.1021/ja202799r

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Traceless and Site-Specific Ubiquitination of Recombinant Proteins

Journal article published in 2011 by Satpal Virdee, Prashant B. Kapadnis, Thomas Elliott, Kathrin Lang ORCID, Julia Madrzak, Duy P. Nguyen, Lutz Riechmann, Jason W. Chin
This paper is available in a repository.
This paper is available in a repository.

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Abstract

Protein ubiquitination is a post-translational modification that regulates almost all aspects of eukaryotic biology. Here we discover the first routes for the efficient site-specific incorporation of delta-thiol-L-lysine (7) and delta-hydroxy-L-lysine (8) into recombinant proteins, via evolution of a pyrrolysyl-tRNA synthetase/tRNA(CUA) pair. We combine the genetically directed incorporation of 7 with native chemical ligation and desulfurization to yield an entirely native isopeptide bond between substrate proteins and ubiquitin. We exemplify this approach by demonstrating the synthesis of a ubiquitin dimer and the first synthesis of ubiquitinated SUMO.