Biosynthesis of a Site-Specific DNA Cleaving Protein
An E. coli catabolite activator protein (CAP) has been converted into a sequence-specific DNA cleaving protein by genetically introducing (2,2′-bipyridin-5-yl)alanine (Bpy-Ala) into the protein. The mutant CAP (CAP-K26Bpy-Ala) showed comparable binding affinity to CAP-WT for the consensus operator sequence. In the presence of Cu(II) and 3-mercaptopropionic acid, CAP-K26Bpy-Ala cleaves double-stranded DNA with high sequence specificity. This method should provide a useful tool for mapping the molecular details of protein−nucleic acid interactions.
Metal: CuLigand type: BipyridineHost protein: Catabolite activator protein (CAP)Anchoring strategy: ---Optimization: Chemical & geneticMax TON: ---ee: ---PDB: ---Notes: Catabolite activator protein from E. coli