Site-specific labeling of proteins with NMR-active unnatural amino acids

J Biomol NMR. 2010 Jan;46(1):89-100. doi: 10.1007/s10858-009-9365-4. Epub 2009 Aug 9.

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.

Publication types

  • Review

MeSH terms

  • Amino Acids / chemistry*
  • Carbon Isotopes / chemistry
  • Codon, Terminator
  • Fluorine / chemistry
  • Isotope Labeling / methods*
  • Nitrogen Isotopes / chemistry
  • Nuclear Magnetic Resonance, Biomolecular / methods*
  • Prokaryotic Cells / chemistry
  • Protein Conformation*
  • Proteins / chemistry*
  • Spin Labels

Substances

  • Amino Acids
  • Carbon Isotopes
  • Codon, Terminator
  • Nitrogen Isotopes
  • Proteins
  • Spin Labels
  • Fluorine