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.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Achenbach J,Jahnz M,Bethge L,Paal K,Jung M,Schuster M,Albrecht R,Jarosch F,Nierhaus KH,Klussmann Sdoi
10.1093/nar/gkv566subject
Has Abstractpub_date
2015-07-13 00:00:00pages
5687-98issue
12eissn
0305-1048issn
1362-4962pii
gkv566journal_volume
43pub_type
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