Traceless and site-specific ubiquitination of recombinant proteins

Satpal Virdee, Prashant B. Kapadnis, Thomas Elliott, Kathrin Lang, Julia Madrzak, Duy P. Nguyen, Lutz Riechmann, Jason W. Chin (Lead / Corresponding author)

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    113 Citations (Scopus)

    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.

    Original languageEnglish
    Pages (from-to)10708-10711
    Number of pages4
    JournalJournal of the American Chemical Society
    Volume133
    Issue number28
    DOIs
    Publication statusPublished - 20 Jul 2011

    Cite this

    Virdee, S., Kapadnis, P. B., Elliott, T., Lang, K., Madrzak, J., Nguyen, D. P., Riechmann, L., & Chin, J. W. (2011). Traceless and site-specific ubiquitination of recombinant proteins. Journal of the American Chemical Society, 133(28), 10708-10711. https://doi.org/10.1021/ja202799r