Crystallization of rat procathepsin B.

  title={Crystallization of rat procathepsin B.},
  author={Jayanthi Sivaraman and R Coloumbe and Michel Magny and Peter Howard Mason and John S. Mort and Miroslaw Cygler},
  journal={Acta crystallographica. Section D, Biological crystallography},
  volume={52 Pt 4},
Rat procathepsin B has been expressed in the yeast Pichia pastoris. To facilitate crystallization of the proform two mutations were introduced: Cys29Ser to avoid self-processing and Ser115Ala to eliminate an N-glycosylation site. The recombinant protein was purified and crystallized by vapor diffusion against mother liquor containing 100 mM KSCN, 100 mM phosphate buffer, pH 6.5 and polyethylene glycol (PEG) 3350 as a precipitating agent. Crystal size was increased by multiple macroseeding. At a… CONTINUE READING

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