Rapid reaction analysis of the catalytic cycle of the EcoRV restriction endonuclease.

@article{Baldwin1995RapidRA,
  title={Rapid reaction analysis of the catalytic cycle of the EcoRV restriction endonuclease.},
  author={Graham S Baldwin and I Barry Vipond and Stephen E. Halford},
  journal={Biochemistry},
  year={1995},
  volume={34 2},
  pages={705-14}
}
We have used the intrinsic tryptophan fluorescence of the EcoRV restriction endonuclease to monitor changes in protein conformation during binding and cleavage of a duplex oligodeoxynucleotide substrate. Appropriate conditions for single-turnover reactions were first determined by steady-state kinetics. When single turnovers were monitored by stopped-flow fluorescence, the mixing together of EcoRV, oligonucleotide and MgCl2 resulted in a rapid increase in tryptophan fluorescence followed by a… CONTINUE READING
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