title: Orchestrating the biosynthesis of an unnatural pyrrolysine amino Acid for its direct incorporation into proteins inside living cells.
abstract: We here report the construction of an E. coli expression system able to manufacture an unnatural amino acid by artificial biosynthesis. This can be orchestrated with incorporation into protein by amber stop codon suppression inside a living cell. In our case an alkyne-bearing pyrrolysine amino acid was biosynthesized and incorporated site-specifically allowing orthogonal double protein labeling.
authors.affiliation: Center for Integrative Protein Science at the Department of Chemistry, Ludwig-Maximilians Universität München, Butenandtstrasse 5-13, 81377 Munich (Germany).
authors.forename: Michael
authors.initials: M
authors.lastname: Ehrlich
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Gattner
authors.forename: Bastien
authors.initials: B
authors.lastname: Viverge
authors.forename: Johanna
authors.initials: J
authors.lastname: Bretzler
authors.forename: David
authors.initials: D
authors.lastname: Eisen
authors.forename: Michael
authors.initials: M
authors.lastname: Stadlmeier
authors.forename: Milan
authors.initials: M
authors.lastname: Vrabel
authors.forename: Thomas
authors.initials: T
authors.lastname: Carell
mesh_terms: D000595:Amino Acid Sequence; D053898:Biosynthetic Pathways; D002256:Carbonic Anhydrases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008958:Models, Molecular; D014176:Protein Biosynthesis; D015202:Protein Engineering / Q000379:methods*
chemical_list: D002256:Carbonic Anhydrases; C456839:pyrrolysine; D008239:Lysine
keywords: amber suppression; bioorganic chemistry; pyrrolysine; synthetic biology; unnatural amino acid