A synthetic adenylation-domain-based tRNA-aminoacylation catalyst.
Angew Chem Int Ed Engl
; 54(8): 2492-6, 2015 Feb 16.
Article
em En
| MEDLINE
| ID: mdl-25583137
ABSTRACT
The incorporation of non-proteinogenic amino acids represents a major challenge for the creation of functionalized proteins. The ribosomal pathway is limited to the 20-22 proteinogenic amino acids while nonribosomal peptide synthetases (NRPSs) are able to select from hundreds of different monomers. Introduced herein is a fusion-protein-based design for synthetic tRNA-aminoacylation catalysts based on combining NRPS adenylation domains and a small eukaryotic tRNA-binding domain (Arc1p-C). Using rational design, guided by structural insights and molecular modeling, the adenylation domain PheA was fused with Arc1p-C using flexible linkers and achieved tRNA-aminoacylation with both proteinogenic and non-proteinogenic amino acids. The resulting aminoacyl-tRNAs were functionally validated and the catalysts showed broad substrate specificity towards the acceptor tRNA. Our strategy shows how functional tRNA-aminoacylation catalysts can be created for bridging the ribosomal and nonribosomal worlds. This opens up new avenues for the aminoacylation of tRNAs with functional non-proteinogenic amino acids.
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Base de dados:
MEDLINE
Assunto principal:
Aminoacilação de RNA de Transferência
/
Aminoacil-tRNA Sintetases
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article