Purification and characterization of cathepsin D from herring muscle ( Clupea harengus )

• Main Authors: Nielsen, L.B., Nielsen, Henrik Hauch
• Format: Article in Journal/Newspaper
• Language: English
• Published: 2001
• Subjects: Råvare- og produktteknologi; Antarc*; Antarctic; Icefish
• Online Access: https://orbit.dtu.dk/en/publications/9cdcf5b6-c22e-48cb-9304-8fcb16c90bf8

Cathepsin D was purified and concentrated 469-fold from a homogenate of Clupea harengus muscle. The purified enzyme is a monomer with a molecular weight of 38 000-39 000. It is inhibited by pepstatin and has optimal activity at pH 2.5 with hemoglobin as the substrate. The isoelectric point is at pH 6.8. Glycosidase treatment and binding to Concanavalin A indicated that the enzyme contains one N-linked carbohydrate moiety of the high-mannose type per molecule. The first 21 amino acid residues of the N-terminal showed high similarity to cathepsin D from antarctic icefish liver (Chionodraco hamatus) and trout ovary (Oncorhynchus mykiss). Digestion of the P-chain of oxidized insulin resulted in preferential cleavage at Leu(15)-Tyr(16), (47%), Tyr(16)-Leu(17) (34%) and Ala(14)- Leu(15) (18%). Incubation with myofibrils from herring muscle at pH 4.23 showed that the enzyme mainly degraded myosin, actin and tropomyosin. (C) 2001 Elsevier Science Inc. All rights reserved.