Characterization and Crystallization of Recombinant Human Cathepsin L

Author: Nomura T.   Fujishima A.   Fujisawa Y.  

Publisher: Elsevier

ISSN: 0006-291X

Source: Biochemical and Biophysical Research Communications, Vol.228, Iss.3, 1996-11, pp. : 792-796

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Abstract

Human procathepsin L (25mg) was highly purified from the culture filtrate (4L) of mouse myeloma cells (Sp-HCL/HE14) transformed with human procathepsin L cDNA. The procathepsin L was almost completely converted to the mature form (18mg) under the acidic condition. Some properties of the mature cathepsin L were found to be different from those of the human liver-derived enzyme. In addition, we first produced crystals of mature human cathepsin L with E-64 using polyethylene glycol 6000 as the precipitant. The crystal was orthorhombic and belonged to the space group P2 1 2 1 2 1 . The unit cell dimensions were: a=49.8 A, b=103.9 A, c=47.8 A. The cell volume (2.47 x 10 5 A 3 ) and calculated molecular mass (24.6 kDa) gave a volume/mass ratio of 2.5 A 3 /Da, which indicates that the asymmetric unit contains one molecule of enzyme.

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