title: (19)F-modified proteins and (19)F-containing ligands as tools in solution NMR studies of protein interactions.
abstract: (19)F solution NMR is a powerful and versatile tool to study protein structure and protein-ligand interactions due to the favorable NMR characteristics of the (19)F atom, its absence in naturally occurring biomolecules, and small size. Protocols to introduce (19)F atoms into both proteins and their ligands are readily available and offer the ability to conduct protein-observe (using (19)F-labeled proteins) or ligand-observe (using (19)F-containing ligands) NMR experiments. This chapter provides two protocols for the (19)F-labeling of proteins, using an Escherichia coli expression system: (i) amino acid type-specific incorporation of (19)F-modified amino acids and (ii) site-specific incorporation of (19)F-modified amino acids using recombinantly expressed orthogonal amber tRNA/tRNA synthetase pairs. In addition, we discuss several applications, involving (19)F-modified proteins and (19)F-containing ligands.
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.
authors.forename: Naima G
authors.initials: NG
authors.lastname: Sharaf
authors.affiliation: Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA. Electronic address: amg100@pitt.edu.
authors.forename: Angela M
authors.initials: AM
authors.lastname: Gronenborn
mesh_terms: D024322:Amino Acids, Aromatic / Q000737:chemistry; D005461:Fluorine / Q000737:chemistry*; D007553:Isotope Labeling; D008024:Ligands; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D011506:Proteins / Q000737:chemistry*
chemical_list: D024322:Amino Acids, Aromatic; D008024:Ligands; D011506:Proteins; D005461:Fluorine
keywords: (19)F NMR; (19)F-containing ligands; (19)F-modified proteins; Biosynthetic methods