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Traceless and Site-Specific Ubiquitination of Recombinant Proteins

[Image: see text] 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 δ-thiol-l-lysine (7) and δ-hydroxy-l-lysine (8) into recombinant proteins, via evo...

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Detalles Bibliográficos
Autores principales: Virdee, Satpal, Kapadnis, Prashant B., Elliott, Thomas, Lang, Kathrin, Madrzak, Julia, Nguyen, Duy P., Riechmann, Lutz, Chin, Jason W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2011
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3135006/
https://www.ncbi.nlm.nih.gov/pubmed/21710965
http://dx.doi.org/10.1021/ja202799r
Descripción
Sumario:[Image: see text] 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 δ-thiol-l-lysine (7) and δ-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.