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A highly flexible tRNA acylation method for non-natural polypeptide synthesis

Nature Methods volume 3, pages 357–359 (2006)Cite this article

A Corrigendum to this article was published on 01 August 2006

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Abstract

Here we describe a de novo tRNA acylation system, the flexizyme (Fx) system, for the preparation of acyl tRNAs with nearly unlimited selection of amino and hydroxy acids and tRNAs. The combination of the Fx system with an appropriate cell-free translation system allows us to readily perform mRNA-encoded synthesis of proteins and short polypeptides involving multiple non-natural amino acids.

*Note: In the version of this article initially published, the authors did not declare competing financial interests. They have filed a patent covering some of the information described in the paper and now declare competing financial interests. This error has been corrected in the PDF version of the article.

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Figure 1: Acylation of tRNA by dFx and eFx.
Figure 2: Synthesis of a non-natural peptide by the PURE system.

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  • 19 July 2006

    In the version of this article initially published, the authors did not declare competing financial interests. They have filed a patent covering some of the information described in the paper and now declare competing financial interests. This error has been corrected in the PDF version of the article.

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Acknowledgements

We thank M. Komiyama for the use of MALDI instrumentation. This work was supported by grants from Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research (S) (16101007) and from the US National Institutes of Health (GM59159).

Author information

Authors and Affiliations

  1. Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, 153-8904, Japan

    Hiroshi Murakami, Atsushi Ohta, Hiroshi Ashigai & Hiroaki Suga

  2. Department of Chemistry and Biotechnology, Graduate School of Engineering, The University of Tokyo, Tokyo, 113-8656, Japan

    Atsushi Ohta, Hiroshi Ashigai & Hiroaki Suga

Authors
  1. Hiroshi Murakami
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  2. Atsushi Ohta
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  3. Hiroshi Ashigai
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  4. Hiroaki Suga
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Corresponding author

Correspondence to Hiroaki Suga.

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Supplementary information

Supplementary Fig. 1

Comparison of activity in flexizymes. (PDF 483 kb)

Supplementary Fig. 2

In vitro selection using Hbi-DBE. (PDF 564 kb)

Supplementary Fig. 3

In vitro selection using Phe-CME. (PDF 444 kb)

Supplementary Fig. 4

Fx-independent acylation of tRNA. (PDF 404 kb)

Supplementary Fig. 5

Acid substrates used in this study. (PDF 154 kb)

Supplementary Fig. 6

Site-specific incorporation of various acids into GFP. (PDF 528 kb)

Supplementary Table 1

Yields of acyl-tRNAs. (PDF 548 kb)

Supplementary Methods (DOC 52 kb)

Supplementary Note (DOC 44 kb)

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Murakami, H., Ohta, A., Ashigai, H. et al. A highly flexible tRNA acylation method for non-natural polypeptide synthesis. Nat Methods 3, 357–359 (2006). https://doi.org/10.1038/nmeth877

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