Purification and characterization of cathepsin L-like enzyme from the muscle of anchovy, Engraulis japonica

Comp Biochem Physiol B Biochem Mol Biol. 1997 Nov;118(3):523-9. doi: 10.1016/s0305-0491(97)00181-8.

Abstract

Results relative to inhibitory effects and substrate specificity indicated that a protease from the muscle of anchovy, Engraulis japonica, was a cathepsin L-like enzyme. The enzyme was activated by thiol reagents and inhibited by thiol-blocking reagents. The molecular weight was estimated to be 25.8 kDa by SDS-PAGE. The enzyme exhibited its maximal activity at pH 6.0 and 50 degrees C for casein and N-benzoyl-D, L-arginine-beta-naphthylamide. The enzyme hydrolyzed at the position of Phe1, Asn3, Val13, Glu14, Val19 and Gly24 of the insulin beta-chain. The K'm and kcat of the enzyme were 73.4 microM and 0.5 microM/min, respectively, toward Z-Phe-Arg-MNap.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Animals
  • Cathepsin L
  • Cathepsins / metabolism*
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / isolation & purification*
  • Cysteine Endopeptidases / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology
  • Endopeptidases*
  • Fishes*
  • Hydrogen-Ion Concentration
  • Kinetics
  • Molecular Sequence Data
  • Molecular Weight
  • Muscles / chemistry
  • Muscles / enzymology*
  • Substrate Specificity
  • Temperature

Substances

  • Amino Acids
  • Cysteine Proteinase Inhibitors
  • Cathepsins
  • Endopeptidases
  • Cysteine Endopeptidases
  • Cathepsin L