Identification of putative active-site residues in the DNase domain of colicin E9 by random mutagenesis.

@article{GarinotSchneider1996IdentificationOP,
  title={Identification of putative active-site residues in the DNase domain of colicin E9 by random mutagenesis.},
  author={C Garinot-Schneider and Ansgar J. Pommer and Geoffrey R. Moore and Colin Kleanthous and Richard James},
  journal={Journal of molecular biology},
  year={1996},
  volume={260 5},
  pages={731-42}
}
We have used random mutagenesis to identify putative active-site residues in the C-terminal cytotoxic endonuclease domain of the bacterial toxin colicin E9. Six single-site mutations in the DNase domain were isolated which destroyed the toxic action of the colicin. DNA sequencing identified the mutations as Gly460Asp, Arg544Gly, Glu548Gly, Thr571Ile, His575Tyr and His579Tyr. All six wild-type residues are highly conserved in the DNase domains of both the E group colicins and the closely related… CONTINUE READING