Identification and characterization of the functional amino acids at the active center of pig liver thioltransferase by site-directed mutagenesis.

@article{Yang1991IdentificationAC,
  title={Identification and characterization of the functional amino acids at the active center of pig liver thioltransferase by site-directed mutagenesis.},
  author={Yi F Yang and William Willie Wells},
  journal={The Journal of biological chemistry},
  year={1991},
  volume={266 19},
  pages={12759-65}
}
By using site-directed mutagenesis techniques, the essential amino acids at the catalytic center of porcine thioltransferase (glutaredoxin) were determined. Seven oligonucleotides were designed, synthesized, and used to construct mutants, ETT-C22S, ETT-C25S, ETT-C25A, ETT-R26V, ETT-K27Q, ETT-R26V: K27Q, and ETT-C78S:C82S, by altering their codons in pig liver thioltransferase cDNA/M13mp18 clones. Each of the thioltransferases was purified to homogeneity and its dithiol-disulfide exchange, and… CONTINUE READING

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