Over-expression, Purification, and Characterization of Recombinant Human Arylamine N-Acetyltransferase 1

@article{Wang2005OverexpressionPA,
  title={Over-expression, Purification, and Characterization of Recombinant Human Arylamine N-Acetyltransferase 1},
  author={Haiqing Wang and Gregory M. Vath and Akane Kawamura and Caleb A Bates and Edith Sim and Patrick E. Hanna and Carston R Wagner},
  journal={The Protein Journal},
  year={2005},
  volume={24},
  pages={65-77}
}
Human arylamine N-acetyltransferase 1 (NAT1) has been overexpressed in E. coli as a mutant dihydrofolic acid reductase (DHFR) fusion protein with a thrombin sensitive linker. An initial DEAE anion-exchange chromatography resulted in partial purification of the fusion protein. The fusion protein was cleaved with thrombin, and human rNAT1 was purified with a second DEAE column. A total of 8 mg of human rNAT1 from 2 l of cell culture was purified to homogeneity with this methodology. Arylamine… Expand
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The new purification methodology utilizes the unique pH dependence of binding to a methotrexate (MTX)-affinity column by the L54F DHFR mutant, and although the specific activity of the purified rNAT2 was comparable to that of NAT2 obtained from hamster tissue, only 3% of the activity was recovered. Expand
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TLDR
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