title: Site-specific labeling of proteins with NMR-active unnatural amino acids.
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.
authors: Genomics Institute of the Novartis Research Foundation, San Diego, CA 92121-1125, USA.
authors: David H
authors: DH
authors: Jones
authors: Susan E
authors: SE
authors: Cellitti
authors: Xueshi
authors: X
authors: Hao
authors: Qiong
authors: Q
authors: Zhang
authors: Michael
authors: M
authors: Jahnz
authors: Daniel
authors: D
authors: Summerer
authors: Peter G
authors: PG
authors: Schultz
authors: Tetsuo
authors: T
authors: Uno
authors: Bernhard H
authors: BH
authors: Geierstanger
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D002247:Carbon Isotopes / Q000737:chemistry; D018388:Codon, Terminator; D005461:Fluorine / Q000737:chemistry; D007553:Isotope Labeling / Q000379:methods*; D009587:Nitrogen Isotopes / Q000737:chemistry; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011387:Prokaryotic Cells / Q000737:chemistry; D011487:Protein Conformation; D011506:Proteins / Q000737:chemistry*; D013113:Spin Labels
chemical_list: D000596:Amino Acids; D002247:Carbon Isotopes; D018388:Codon, Terminator; D009587:Nitrogen Isotopes; D011506:Proteins; D013113:Spin Labels; D005461:Fluorine