Outwitting EF-Tu and the ribosome: translation with D-amino acids

被引：49

作者：

Achenbach, John ^{[1
]}

Jahnz, Michael ^{[1
]}

Bethge, Lucas ^{[1
]}

Paal, Krisztina ^{[1
]}

Jung, Maria ^{[1
]}

Schuster, Maja ^{[1
]}

Albrecht, Renate ^{[2
]}

Jarosch, Florian ^{[1
]}

Nierhaus, Knud H. ^{[2
]}

Klussmann, Sven ^{[1
]}

机构：

[1] NOXXON Pharma AG, D-10589 Berlin, Germany

[2] Charite, Inst Med Phys & Biophys, D-10117 Berlin, Germany

来源：

NUCLEIC ACIDS RESEARCH | 2015年 / 43卷 / 12期

关键词：

ELONGATION-FACTOR-TU; SITE-SPECIFIC INCORPORATION; PEPTIDE-BOND FORMATION; IN-VITRO SELECTION; TRANSFER-RNA; ESCHERICHIA-COLI; PROTEIN-BIOSYNTHESIS; NONNATURAL RESIDUES; UNIFORM BINDING; AFFINITY;

D O I：

10.1093/nar/gkv566

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 tRNAGly 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.

引用

页码：5687 / 5698

页数：12

共 65 条

[61] DISCRIMINATION BETWEEN D-TYROSYL AND L-TYROSYL TRANSFER RIBONUCLEIC-ACIDS IN PEPTIDE-CHAIN ELONGATION [J].