A genetically encoded fluorescent amino acid

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2006
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Chen, Shuo
Wu, Ning
Deiters, Alexander
Chin, Jason W.
Schultz, Peter G.
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Zusammenfassung

The ability to introduce fluorophores selectively into proteins provides a powerful tool to study protein structure, dynamics, localization, and biomolecular interactions both in vitro and in vivo. Here, we report a strategy for the selective and efficient biosynthetic incorporation of a low-molecular-weight fluorophore into proteins at defined sites. The fluorescent amino acid 2-amino-3-(5-(dimethylamino)naphthalene-1-sulfonamide)propanoic acid (dansylalanine) was genetically encoded in Saccharomyces cerevisiae by using an amber nonsense codon and corresponding orthogonal tRNA/aminoacyl-tRNA synthetase pair. This environmentally sensitive fluorophore was selectively introduced into human superoxide dismutase and used to monitor unfolding of the protein in the presence of guanidinium chloride. The strategy described here should be applicable to a number of different fluorophores in both prokaryotic and eukaryotic organisms, and it should facilitate both biochemical and cellular studies of protein structure and function.

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ISO 690SUMMERER, Daniel, Shuo CHEN, Ning WU, Alexander DEITERS, Jason W. CHIN, Peter G. SCHULTZ, 2006. A genetically encoded fluorescent amino acid. In: Proceedings of the National Academy of Sciences. 2006, 103(26), pp. 9785-9789. ISSN 0027-8424. Available under: doi: 10.1073/pnas.0603965103
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@article{Summerer2006-06-27genet-385,
  year={2006},
  doi={10.1073/pnas.0603965103},
  title={A genetically encoded fluorescent amino acid},
  number={26},
  volume={103},
  issn={0027-8424},
  journal={Proceedings of the National Academy of Sciences},
  pages={9785--9789},
  author={Summerer, Daniel and Chen, Shuo and Wu, Ning and Deiters, Alexander and Chin, Jason W. and Schultz, Peter G.}
}
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