title: Outwitting EF-Tu and the ribosome: translation with d-amino acids.
abstract: Key components of the translational apparatus, i.e. ribosomes, elongation factor EF-Tu and most aminoacyl-tRNA synthetases, are stereoselective and prevent incorporation of d-amino acids (d-aa) into polypeptides. The rare appearance of d-aa in natural polypeptides arises from post-translational modifications or non-ribosomal synthesis. We introduce an in vitro translation system that enables single incorporation of 17 out of 18 tested d-aa into a polypeptide; incorporation of two or three successive d-aa was also observed in several cases. The system consists of wild-type components and d-aa are introduced via artificially charged, unmodified tRNA(Gly) that was selected according to the rules of 'thermodynamic compensation'. The results reveal an unexpected plasticity of the ribosomal peptidyltransferase center and thus shed new light on the mechanism of chiral discrimination during translation. Furthermore, ribosomal incorporation of d-aa into polypeptides may greatly expand the armamentarium of in vitro translation towards the identification of peptides and proteins with new properties and functions.
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany jachenbach@noxxon.com.
authors.forename: John
authors.identifier: http://orcid.org/0000-0001-8161-4335
authors.initials: J
authors.lastname: Achenbach
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Jahnz
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Lucas
authors.initials: L
authors.lastname: Bethge
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Krisztina
authors.initials: K
authors.lastname: Paal
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Jung
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Maja
authors.initials: M
authors.lastname: Schuster
authors.affiliation: Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany.
authors.forename: Renate
authors.initials: R
authors.lastname: Albrecht
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Jarosch
authors.affiliation: Institut für Medizinische Physik und Biophysik, Charité - Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany nierhaus@molgen.mpg.de.
authors.forename: Knud H
authors.initials: KH
authors.lastname: Nierhaus
authors.affiliation: NOXXON Pharma AG, Max-Dohrn-Strasse 8-10, 10589 Berlin, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Klussmann
mesh_terms: D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism; D010452:Peptide Biosynthesis; D010444:Peptide Elongation Factor Tu / Q000737:chemistry / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D012343:RNA, Transfer / Q000737:chemistry / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry / Q000378:metabolism*; D013237:Stereoisomerism; D046249:Transfer RNA Aminoacylation
chemical_list: D000596:Amino Acids; D010455:Peptides; D012346:RNA, Transfer, Amino Acyl; D012343:RNA, Transfer; D010444:Peptide Elongation Factor Tu