Document Detail


In situ chemical aminoacylation with amino acid thioesters linked to a peptide nucleic acid.
MedLine Citation:
PMID:  15584731     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
tRNA-specific chemical aminoacylation was achieved with nonnatural amino acids. A nonnatural amino acid was activated as a thioester derivative, and the latter was linked through a spacer to the N-terminal of a 9-mer peptide nucleic acid that is complementary to the 3'-terminal region of yeast phenylalanine tRNA. Efficient aminoacylation was observed when the amino acid thioester-spacer-PNA conjugate was mixed with the tRNA. The PNA-assisted aminoacylation was also successful in an Escherichia coli in vitro protein synthesizing system that contained an orthogonalized tRNA. The in situ aminoacylation/in vitro translation gave a mutant protein in which the nonnatural amino acid was incorporated into the position directed by a CGGG 4-base codon/anticodon pair.
Authors:
Keiko Ninomiya; Toshikazu Minohata; Masaki Nishimura; Masahiko Sisido
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  126     ISSN:  0002-7863     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2004 Dec 
Date Detail:
Created Date:  2004-12-08     Completed Date:  2005-02-15     Revised Date:  2012-08-01    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  15984-9     Citation Subset:  IM    
Affiliation:
Department of Bioscience and Biotechnology, Faculty of Engineering, Okayama University, 3-1-1 Tsushimanaka, Okayama 700-8530, Japan.
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MeSH Terms
Descriptor/Qualifier:
Acylation
Amino Acids / chemistry*,  metabolism
Computer-Aided Design
Escherichia coli / genetics,  metabolism
Esters / chemistry,  metabolism
Models, Molecular
Nucleic Acid Hybridization
Peptide Nucleic Acids / biosynthesis,  chemistry*
Protein Biosynthesis*
RNA, Transfer, Amino Acyl / chemistry*,  metabolism
Sulfhydryl Compounds / chemistry,  metabolism
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Esters; 0/Peptide Nucleic Acids; 0/RNA, Transfer, Amino Acyl; 0/Sulfhydryl Compounds

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