Site-specific labeling of proteins with NMR-active unnatural amino acids.
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.
Jones
David H
DH
Genomics Institute of the Novartis Research Foundation, San Diego, CA 92121-1125, USA.
Cellitti
Susan E
SE
Hao
Xueshi
X
Zhang
Qiong
Q
Jahnz
Michael
M
Summerer
Daniel
D
Schultz
Peter G
PG
Uno
Tetsuo
T
Geierstanger
Bernhard H
BH
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
D000596:Amino Acids; D002247:Carbon Isotopes; D018388:Codon, Terminator; D009587:Nitrogen Isotopes; D011506:Proteins; D013113:Spin Labels; D005461:Fluorine