Abstract A large number of amino acids other than the canonical amino acids can now be easily incorporated in vivo into proteins at genetically encoded positions. The technology requires an orthogonal tRNA/aminoacyl-tRNA synthetase pair specific for the unnatural amino acid that is added to the media while a TAG amber or frame shift codon specifies the incorporation site in the protein to be studied. These unnatural amino acids can be isotopically labeled and provide unique opportunities for site-specific labeling of proteins for NMR studies. In this perspective, we discuss these opportunities including new photocaged unnatural amino acids, outline usage of metal chelating and spin-labeled unnatural amino acids and expand the approach to in-cell NMR experiments.
Content Type Journal Article
Pages 89-100
DOI 10.1007/s10858-009-9365-4
Authors
David H. Jones, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
Susan E. Cellitti, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
Xueshi Hao, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
Qiong Zhang, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
Michael Jahnz, The Scripps Research Institute Department of Chemistry and the Skaggs Institute for Chemical Biology 10550 North Torrey Pines Road La Jolla CA 92037 USA
Daniel Summerer, The Scripps Research Institute Department of Chemistry and the Skaggs Institute for Chemical Biology 10550 North Torrey Pines Road La Jolla CA 92037 USA
Peter G. Schultz, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
Tetsuo Uno, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
Bernhard H. Geierstanger, Genomics Institute of the Novartis Research Foundation 10675 John Jay Hopkins Drive San Diego CA 92121-1125 USA
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Site-specific labeling of proteins with NMR-active unnatural amino acids
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