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Purification and partial characterization of cathepsin D from porcine (Sus scrofa) liver using affinity
F Canduri1, R J Ward, W F de Azevedo Júnior
1Departamento de Física, IBILCE/UNESP, São José do Rio Preto, Brazil.
Biochemistry and Molecular Biology International
|August 26, 1998
Abstract:
Cathepsin D, a lysosomal aspartic protease, has been purified from porcine liver using a combination of pepstatin-A agarose and Affi-Gel Blue affinity chromatography, followed by size-exclusion chromatography. The purified protein consists of two polypeptide chains of 15 and 30 kDa, and has an isoelectric point of 6.8. Porcine liver cathepsin D has maximum activity at pH 2.5-3.0 as determined by its activity against hemoglobin, with a Kcat of 14.3 s-1 and a kcat/KM of 2.70 x 10(6) s-1M-1 as determined by the hydrolysis of a fluorogenic peptide substrate.