Site-specific incorporation of (aminooxy)acetic acid into proteins

Brian M. Eisenhauer, Sidney M. Hecht

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

By employing a general biosynthetic method for the elaboration of proteins containing unnatural amino acid analogues, we incorporated (aminooxy)acetic acid into positions 10 and 27 of Escherichia coli dihydrofolate reductase. Introduction of the modified amino acid into DHFR was accomplished in an in vitro protein biosynthesizing system by readthrough of a nonsense (UAG) codon with a suppressor tRNA that had been activated with (aminooxy)acetic acid. Incorporation of the amino acid proceeded with reasonable efficiency at codon position 10 but less well at position 27. (Aminooxy)acetic acid was also incorporated into position 72 of DNA polymerase β. Peptides containing (aminooxy)acetic acid have been shown to adopt a preferred conformation involving an eight-membered ring that resembles a γ-turn. Accordingly, the present study may faciliate the elaboration of proteins containing conformationally biased peptidomimetic motifs at predetermined sites. The present results further extend the examples of ribosomally mediated formation of peptide bond analogues of altered connectivity and provide a conformationally biased linkage at a predetermined site. It has also been shown that the elaborated protein can be cleaved chemically at the site containing the modified amino acid.

Original languageEnglish (US)
Pages (from-to)11472-11478
Number of pages7
JournalBiochemistry
Volume41
Issue number38
DOIs
StatePublished - Sep 24 2002
Externally publishedYes

ASJC Scopus subject areas

  • Biochemistry

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