title: Protein-DNA photo-crosslinking with a genetically encoded benzophenone-containing amino acid.
abstract: The photo-crosslinking amino acid, p-benzoyl-L-phenylalanine (pBpa), was genetically incorporated into Escherichia coli catabolite activator protein (CAP) in bacteria in response to an amber nonsense codon using an orthogonal tRNA/aminoacyl-tRNA synthetase pair. The mutant CAP (CAP-K26Bpa) containing pBpa formed a covalent complex with a DNA fragment containing the consensus operator sequence upon UV irradiation. Because this amino acid can be genetically incorporated into any DNA-binding protein in E. coli, yeast or mammalian cells with minimal perturbation of protein structure, this method should be generally useful for investigating DNA-protein interactions.
authors: Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors: Hyun Soo
authors: HS
authors: Lee
authors: Romerson D
authors: RD
authors: Dimla
authors: Peter G
authors: PG
authors: Schultz
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D000604:Amino Acyl-tRNA Synthetases / Q000378:metabolism; D001483:Base Sequence; D001577:Benzophenones / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D002373:Cyclic AMP Receptor Protein / Q000737:chemistry* / Q000378:metabolism; D004247:DNA / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D008969:Molecular Sequence Data; D011485:Protein Binding; D012343:RNA, Transfer / Q000378:metabolism; D014466:Ultraviolet Rays
chemical_list: D000596:Amino Acids; D001577:Benzophenones; D003432:Cross-Linking Reagents; D002373:Cyclic AMP Receptor Protein; C047723:benzophenone; D004247:DNA; D012343:RNA, Transfer; D000604:Amino Acyl-tRNA Synthetases