New method to discriminate between cathepsin B and cathepsin L in crude extracts from fish muscle based on a simple acidification procedure

Helene Godiksen, Henrik Hauch Nielsen

Research output: Contribution to journalJournal articlepeer-review

Abstract

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.
Original languageEnglish
JournalInternational Journal of Food Science and Technology
Volume42
Issue number1
Pages (from-to)102-106
ISSN0950-5423
DOIs
Publication statusPublished - 2007

Fingerprint

Dive into the research topics of 'New method to discriminate between cathepsin B and cathepsin L in crude extracts from fish muscle based on a simple acidification procedure'. Together they form a unique fingerprint.

Cite this