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Published online on June 19, 2006, 10.1073/pnas.0603965103
PNAS | June 27, 2006 | vol. 103 | no. 26 | 9785-9789


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BIOLOGICAL SCIENCES / BIOCHEMISTRY
A genetically encoded fluorescent amino acid

Daniel Summerer*, Shuo Chen*, Ning Wu{dagger}, Alexander Deiters{ddagger}, Jason W. Chin§, and Peter G. Schultz*,§

*Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, SR202, La Jolla, CA 92037; {dagger}Beth Israel Deaconess Medical Center, Division of Signal Transduction, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02115; {ddagger}Department of Chemistry, North Carolina State University, Campus Box 8240, Raleigh, NC 27695; and §Division of Protein and Nucleic Acid Chemistry, Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom

Contributed by Peter G. Schultz, May 12, 2006

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.

molecular evolution | fluorescent probes | genetic code expansion | protein design | unnatural amino acids

Author contributions: D.S., S.C., N.W., and P.G.S. designed research; D.S., S.C., and N.W. performed research; A.D. and J.W.C. contributed new reagents/analytical tools; D.S., N.W., and P.G.S. analyzed data; and D.S. and P.G.S. wrote the paper.

Conflict of interest statement: No conflicts declared.

To whom correspondence should be addressed. E-mail: schultz{at}scripps.edu

© 2006 by The National Academy of Sciences of the USA


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