Improved purification protocol for wild-type and mutant human foamy virus proteases.

@article{Boross2006ImprovedPP,
  title={Improved purification protocol for wild-type and mutant human foamy virus proteases.},
  author={P{\'e}ter Boross and J{\'o}zsef T{\"o}zs{\'e}r and P{\'e}ter Bagossi},
  journal={Protein expression and purification},
  year={2006},
  volume={46 2},
  pages={343-7}
}
Wild-type and an active site mutant (S25T) human foamy virus (HFV) proteases were expressed in fusion with maltose binding protein in Escherichia coli. The mutant enzyme contained a Ser to Thr mutation in the -Asp-Ser-Gly- active site triplet of the enzyme, which forms the "fireman's grip" between the two subunits of the homodimeric enzyme. The fusion proteins were purified by affinity chromatography on amylose resin, cleaved with factor Xa, and the processed enzymes were purified by gel… CONTINUE READING