Purification and crystallization of rhizopuspepsin: the use of nickel chelation chromatography to select for catalytically active species.
@article{Flentke1999PurificationAC, title={Purification and crystallization of rhizopuspepsin: the use of nickel chelation chromatography to select for catalytically active species.}, author={G. R. Flentke and J. Gliński and K. Satyshur and D. Rich}, journal={Protein expression and purification}, year={1999}, volume={16 2}, pages={ 213-20 } }
A new method to obtain pure zymogen-derived peptidases is presented. The key strategy is to install a polyhistidine peptide tag on the N-terminus of the propeptide sequence of a zymogen. After expression, purification, and folding of the protein, autocatalytic posttranslational cleavage and filtration through a nickel affinity column gives pure, functional peptidase. This method takes advantage of the nickel affinity chromatography system that removes both zymogen peptide and nonfunctional… CONTINUE READING
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