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Ribosome-mediated incorporation of a non-standard amino acid into a peptide through expansion of the genetic code

Nature volume 356, pages 537–539 (1992)Cite this article

Abstract

ONE serious limitation facing protein engineers is the availability of only 20 'proteinogenic' amino acids encoded by natural messenger RNA. The lack of structural diversity among these amino acids restricts the mechanistic and structural issues that can be addressed by site-directed mutagenesis. Here we describe a new technology for incorporating non-standard amino acids into polypeptides by ribosome-based translation. In this technology, the genetic code is expanded through the creation of a 65th codon-anticodon pair from unnatural nucleoside bases having non-standard hydrogen-bonding patterns1,2. This new codon-anticodon pair efficiently supports translation in vitro to yield peptides containing a non-standard amino acid. The versatility of the ribo-some as a synthetic tool offers new possibilities for protein engineering, and compares favourably with another recently described approach in which the genetic code is simply rearranged to recruit stop codons to play a coding role3–9. Spelling of author S.A.B. was fixed in the HTML on 2 February 2016.

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References

  1. Switzer, C. Y., Moroney, S. E. & Benner, S. A. J. Am. chem. Soc. 111, 8322–8323 (1989).

    Article  CAS  Google Scholar 

  2. Piccirilli, J. A., Krauch, T., Moroney, S. E. & Benner, S. A. Nature 343, 33–37 (1990).

    Article  ADS  CAS  Google Scholar 

  3. Bain, J. D., Glabe, C. G., Dix, T. A., Chamberlin, A. R., & Diala, E, S. J. Am. chem. Soc. 111, 8013–8014 (1989).

    Article  CAS  Google Scholar 

  4. Bain, J. D., Wacker, D. A., Kuo, E. E. & Chamberlin, A. R. Tetrahedron 14/15, 2389–2400 (1991).

    Article  Google Scholar 

  5. Hecht, S. M., Alford, B. L., Kuroda, Y., & Kitano, S. J. biol. Chem. 253, 4517–4520 (1978).

    CAS  PubMed  Google Scholar 

  6. Roesser, J. R., Chorghade, M. S. & Hecht, S. M. Biochemistry 25, 6361–6365 (1986).

    Article  CAS  Google Scholar 

  7. Baldini, G., Martoglio, B., Schachenmann, A., Zugliani, C. & Brunner, J. Biochemistry 27, 7951–7959 (1988).

    Article  CAS  Google Scholar 

  8. Noren, C. J., Anthony-Cahill, S. J., Griffith, M. C. & Schultz, P. G. Science 244, 182–188 (1989).

    Article  ADS  CAS  Google Scholar 

  9. Robertson, S. A., Ellman, J. A. & Schultz, P. G. J. Am. chem. Soc. 113, 2722–2729 (1991).

    Article  CAS  Google Scholar 

  10. Kozak, M. Nature 308, 241–246 (1984).

    Article  ADS  CAS  Google Scholar 

  11. Mantsch, H. H. et al. Biochemistry 14, 5593–5601 (1975).

    Article  CAS  Google Scholar 

  12. McBride, L. J., Kierzek, R., Beaucage, S. L. & Caruthers, M. H. J. Am. chem. Soc. 108, 2040–2048 (1986).

    Article  CAS  Google Scholar 

  13. Bain, J. D. et al. Biochemistry 30, 5411–5421 (1991).

    Article  CAS  Google Scholar 

  14. Fluck, M. M., Salser, W. & Epstein, R. H. Molec gen. Genet. 151, 137–149 (1977).

    Article  CAS  Google Scholar 

  15. Konecki, D. S., Aune, K. C., Tate, W. & Caskey, C. T. J. biol. Chem. 252, 4514–4520 (1977).

    CAS  PubMed  Google Scholar 

  16. Smrt, J., Kemper, W., Caskey, T. & Nirenberg, M. J. biol. Chem. 245, 2753–2757 (1970).

    CAS  PubMed  Google Scholar 

  17. Valle, R. P. C. & Morch, M.-D. FEBS Lett. 235, 1–15 (1988).

    Article  CAS  Google Scholar 

  18. Craigen, W. J. & Caskey, C. T. Cell, 50, 1–2 (1987).

    Article  CAS  Google Scholar 

  19. Tsuchihashi, Z. & Kornberg, A. Proc. natn. Acad. Sci U.S.A. 87, 2516–2520 (1990).

    Article  ADS  CAS  Google Scholar 

  20. Jukes, T. J., Bessho, Y., Ohama, T. & Osawa, S. Nature 352, 575 (1991).

    Article  ADS  CAS  Google Scholar 

  21. Fersht, A. R. Biochemistry 26, 8031–8037 (1987).

    Article  CAS  Google Scholar 

  22. Kast, P. thesis ETH, Zurich (1991).

  23. Roberts, J. W. & Carbon, J. J. biol. Chem. 250, 5530–5541 (1975).

    CAS  PubMed  Google Scholar 

  24. Lyttle, M. H., Wright, P. B., Sinha, N. D., Bain, J. D. & Chamberlin, A. R. J. org. Chem. 56, 4608–4615 (1991).

    Article  CAS  Google Scholar 

  25. Bain, J. D., Wacker, D. A., Kuo, E. E., Lyttle, M. H. & Chamberlin, A. R. J. org. Chem. 56, 4615–4625 (1991).

    Article  CAS  Google Scholar 

  26. Fersht, A. R. & Dingwall, C. Biochemistry 18, 2627–2631 (1979).

    Article  CAS  Google Scholar 

  27. Sepiol, J., Kazimierczuk, Z. & Shugar, D. Z Naturf. 31c, 361–370 (1976).

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Chemistry, University of California, irvine, California, 92717, USA

    J. D. Bain & Richard Chamberlin

  2. Department of Chemistry, University of California, Riverside, California, 9251, USA

    Christopher Switzer

  3. Laboratory for Organic Chemistry, Eidgenössische Technische Hochschule, Zurich, CH-8092, Switzerland

    Steven A. Benner

Authors
  1. J. D. Bain
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  2. Christopher Switzer
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  3. Richard Chamberlin
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  4. Steven A. Benner
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Bain, J., Switzer, C., Chamberlin, R. et al. Ribosome-mediated incorporation of a non-standard amino acid into a peptide through expansion of the genetic code. Nature 356, 537–539 (1992). https://doi.org/10.1038/356537a0

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