Journal article
New method to discriminate between cathepsin B and cathepsin L in crude extracts from fish muscle based on a simple acidification procedure
A new and simple method to distinguish between cathepsin B and cathepsin L in crude extracts of herring (Clupea harengus) muscle has been established. An acid treatment of crude extracts (exposed to pH 3 for 5 min) activated a latent form of cathepsin L and inactivated cathepsin B. Furthermore, in neutral crude extract, the hydrolysis of benzyloxycarbonyl-L-phenylalanyl-L-arginyl-4-methylcoumarine (Z-Phe-Arg-MCA) (cathepsin B and cathepsin L substrates) was between 0% and 15% of the hydrolysis of benzyloxycarbonyl-L-arginyl-L-arginyl-7-amino-4-methylcoumarine (Z-Arg-Arg-MCA; cathepsin B substrate).
Cathepsin B activity is measured in neutral extract using the specific cathepsin B substrate Z-Arg-Arg-MCA and cathepsin L activity is measured in acid-treated extract with Z-Phe-Arg-MCA as substrate. The specific cathepsin B inhibitor, CA-074, did not inhibit the Z-Arg-Arg-MCA significantly without affecting the Z-Phe-Arg-MCA activity.
An acid treatment of the crude extract is therefore a more advantageous approach to discriminate between cathepsin B and cathepsin L activities.
Language: | English |
---|---|
Year: | 2007 |
Pages: | 102-106 |
ISSN: | 13652621 and 09505423 |
Types: | Journal article |
DOI: | 10.1111/j.1365-2621.2006.01254.x |