Active site of triosephosphate isomerase: in vitro mutagenesis and characterization of an altered enzyme.

@article{Straus1985ActiveSO,
  title={Active site of triosephosphate isomerase: in vitro mutagenesis and characterization of an altered enzyme.},
  author={David M. Straus and Ronald T. Raines and E. Kawashima and Jeremy Knowles and Walter Gilbert},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={1985},
  volume={82 8},
  pages={2272-6}
}
We have replaced the glutamic acid-165 at the active site of chicken triosephosphate isomerase with an aspartic acid residue using site-directed mutagenesis. Expression of the mutant protein in a strain of Escherichia coli that lacks the bacterial isomerase results in a complementation phenotype that is intermediate between strains that have no isomerase and strains that produce either the wild-type chicken enzyme or the native E. coli isomerase. The value of kcat for the purified mutant enzyme… CONTINUE READING