Protein-DNA photo-crosslinking with a genetically encoded benzophenone-containing amino acid.

Hyun Soo Lee, Romerson D Dimla, Peter G Schultz

Department of Chemistry and the Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.

Bioorganic & medicinal chemistry letters 2009 Sep 1

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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.

Citation

Hyun Soo Lee, Romerson D Dimla, Peter G Schultz. Protein-DNA photo-crosslinking with a genetically encoded benzophenone-containing amino acid. Bioorganic & medicinal chemistry letters. 2009 Sep 1;19(17):5222-4