Elongation factor Tu mutants expand amino acid tolerance of protein biosynthesis system

Yoshio Doi, Takashi Ohtsuki, Yoshihiro Shimizu, Takuya Ueda, Masahiko Sisido

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Nonnatural amino acids have been introduced into proteins using expanded protein biosynthesis systems. However, some nonnatural amino acids, especially those containing large aromatic groups, are not efficiently incorporated into proteins. Reduced binding efficiency of aminoacylated tRNAs to elongation factor Tu (EF-Tu) is likely to limit incorporation of large amino acids. Our previous studies suggested that tRNAs carrying large nonnatural amino acids are bound less tightly to EF-Tu than natural amino acids. To expand the availability of nonnatural mutagenesis, EF-Tu from the E. coli translation system was improved to accept such large amino acids. We synthesized EF-Tu mutants, in which the binding pocket of the aminoacyl moiety of aminoacyl-tRNA was enlarged. L-1-Pyrenylalanine, L-2-pyrenylalanine, and DL-2-anthraquinonylalanine, which are hardly or only slightly incorporated with the wild-type EF-Tu, were successfully incorporated into a protein using these EF-Tu mutants.

Original languageEnglish
Pages (from-to)14458-14462
Number of pages5
JournalJournal of the American Chemical Society
Volume129
Issue number46
DOIs
Publication statusPublished - Nov 21 2007

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Peptide Elongation Factor Tu
Biosynthesis
Protein Biosynthesis
Amino acids
Elongation
Proteins
Amino Acids
Transfer RNA
Mutagenesis
Escherichia coli
Availability

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Elongation factor Tu mutants expand amino acid tolerance of protein biosynthesis system. / Doi, Yoshio; Ohtsuki, Takashi; Shimizu, Yoshihiro; Ueda, Takuya; Sisido, Masahiko.

In: Journal of the American Chemical Society, Vol. 129, No. 46, 21.11.2007, p. 14458-14462.

Research output: Contribution to journalArticle

Doi, Yoshio ; Ohtsuki, Takashi ; Shimizu, Yoshihiro ; Ueda, Takuya ; Sisido, Masahiko. / Elongation factor Tu mutants expand amino acid tolerance of protein biosynthesis system. In: Journal of the American Chemical Society. 2007 ; Vol. 129, No. 46. pp. 14458-14462.
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