Summary
Cathepsin B activity was found in serum-free spent media of human pancreatic cancer cell lines, Panc-1 and MiaPaca. Cathepsin B activity was partially purified by gel filtration on TSK G3000SW, Con-A Sepharose chromatography, Phenyl-Superose column chromatography, and Mono S column chromatography. The optimal pH of cathepsin B was 7.4, and the activity was retained even at alkaline pH. Heat stability test showed tha the enzyme was heat stable; that is, 50% activity was retained after incubation at 56°C for 60 min. These results suggest that cathepsin B secreted from human pancreatic cancer cell is a variant type and may play an important role in pancreatic cancer invasion or metastasis through destruction of the surrounding extracellular matrix by its proteolytic activity.
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Abbreviations
- Con A:
-
concanavaline A
- kd:
-
kilodalton
- Z-Phe-Phe-CHN2 :
-
Carbobenzoxy-L-Phenylalanyl-L-Phenylalanine CHN2
- Z-Phe-Ala-CHN2 :
-
Carbobenzoxy-L-Phenylalanyl-L-Alanine CHN2
- Z-Arg-Arg-MCA:
-
Carbobenzoxy-L-Arginyl-L-Arginine 4-Methyl-Coumaryl-7-Amide
- Z-Phe-Arg-MCA:
-
Carbobenzoxy-L-Phenylalanyl-L-Arginine 4-Methyl-Coumaryl-7-Amide
- Arg-MCA:
-
L-Arginine 4-Methyl-Coumaryl-7-Amide
- Bz-Arg-MCA:
-
Benzoyl-L-Arginine 4-Methyl-Coumaryl-7-Amide
- AMC:
-
7-Amino-4-Methyl-Coumarine
- SDS-PAGE:
-
Sodium dodecyl sulfate-Polyacrylamide gel electrophoresis
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Chung, S.m., Kawai, K. Variant cathepsin B activity secreted from human pancreatic cancer cell lines into protein-free chemically defined medium. Gastroenterol Jpn 24, 699–706 (1989). https://doi.org/10.1007/BF02774170
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DOI: https://doi.org/10.1007/BF02774170