Engineering subtilisin into a fluoride-triggered processing protease useful for one-step protein purification.

@article{Ruan2004EngineeringSI,
  title={Engineering subtilisin into a fluoride-triggered processing protease useful for one-step protein purification.},
  author={Biao Ruan and Kathryn E Fisher and Patrick A. Alexander and Viktoriya Doroshko and Philip N. Bryan},
  journal={Biochemistry},
  year={2004},
  volume={43 46},
  pages={14539-46}
}
Subtilisin was engineered into a highly specific, processing protease, and the subtilisin prodomain was coengineered into an optimized recognition sequence. This involved five steps. First, a robust subtilisin mutant was created, which could tolerate the subsequent mutations needed for high specificity. Second, the substrate binding pocket was mutated to increase its sequence selectivity. Third, the subtilisin prodomain was engineered to direct cleavage to the junction of any protein fused to… CONTINUE READING