title: Orthogonal Protein Translation Using Pyrrolysyl-tRNA Synthetases for Single- and Multiple-Noncanonical Amino Acid Mutagenesis.
abstract: To date, the two systems most extensively used for noncanonical amino acid (ncAA) incorporation via orthogonal translation are based on the Methanococcus jannaschii TyrRS/tRNACUATyrand the Methanosarcina barkeri/Methanosarcina mazei PylRS/tRNACUAPylpairs. Here, we summarize the development and usage of the pyrrolysine-based system for orthogonal translation, a process that allows for the recombinant production of site-specifically labeled proteins and peptides. Via stop codon suppression in Escherichia coli and mammalian cells, genetically encoded biomolecules can be equipped with a great diversity of chemical functionalities including click chemistry handles, post-translational modifications, and photocaged sidechains.
authors: Department of Chemistry, Berlin Institute of Technology/TU Berlin, Biocatalysis Group, Müller-Breslau-Straße 10 / Sekr. L 1, D-10623, Berlin, Germany.
authors: Tobias
authors: T
authors: Baumann
authors: Department of Chemistry, Berlin Institute of Technology/TU Berlin, Biocatalysis Group, Müller-Breslau-Straße 10 / Sekr. L 1, D-10623, Berlin, Germany.
authors: Matthias
authors: M
authors: Exner
authors: Department of Chemistry, Berlin Institute of Technology/TU Berlin, Biocatalysis Group, Müller-Breslau-Straße 10 / Sekr. L 1, D-10623, Berlin, Germany. budisa@biocat.tu-berlin.de.
authors: Nediljko
authors: N
authors: Budisa
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D018388:Codon, Terminator / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017017:Methanococcus / Q000235:genetics / Q000378:metabolism; D017020:Methanosarcina / Q000235:genetics / Q000378:metabolism; D016296:Mutagenesis; D014176:Protein Biosynthesis / Q000235:genetics*; D013489:Suppression, Genetic / Q000235:genetics*
chemical_list: D001426:Bacterial Proteins; D018388:Codon, Terminator; D000604:Amino Acyl-tRNA Synthetases
keywords: Expanded genetic code; Noncanonical amino acid; Orthogonal translation; Pyrrolysyl-tRNA synthetase; Stop codon suppression