Identification and characterization of a cathepsin L-like cysteine protease from Gnathostoma spinigerum

Mol Biochem Parasitol. 2008 Aug;160(2):129-37. doi: 10.1016/j.molbiopara.2008.05.001. Epub 2008 May 8.


Gnathostoma spinigerum is a causative agent of human gnathostomiasis, a common parasitic disease involving skin and visceral organs, especially the central nervous system. In this study, we identified a cDNA encoding a cathepsin L-like cysteine protease (GsCL1) from the lambdaZAP cDNA library of G. spinigerum advanced third-stage larva (aL3) and characterized the biochemical properties of the recombinant enzyme. The cloned cDNA of 1484bp encoded 398 amino acids which contained a typical signal peptide sequence (23 amino acids), a pro-domain (156 amino acids), and a mature domain (219 amino acids) with an approximate molecular weight of 24kDa. The deduced amino acid sequence of GsCL1 gene showed 53-64% identity to cathepsin L proteases of various organisms including a cathepsin L family member (cpl-1) of Caenorhabditis elegans. Recombinant proGsCL1 expressed in Pichia pastoris showed typical biochemical characteristics of cysteine proteases. The expressed enzyme displayed optimal protease activity toward Z-Phe-Arg-AMC substrate at pH 6.0 but not toward Z-Arg-Arg-AMC. The activity was sensitive to cysteine protease inhibitors E-64 and K11777. The preference for large hydrophilic and aromatic residues in the P2 position (I, L, F, W, U, V) was typical of cathepsin L proteases. Mouse anti-GST-proGsCL1 serum showed reactivity with 35-, 38- and 45-kDa proteins in the aL3 extracts. These proteins were shown to localize inside the intestinal cells of aL3.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Caenorhabditis elegans / enzymology
  • Cathepsin L
  • Cathepsins / chemistry
  • Cathepsins / genetics*
  • Cathepsins / isolation & purification
  • Cathepsins / metabolism*
  • Cloning, Molecular
  • Coumarins / metabolism
  • Cysteine Endopeptidases / chemistry
  • Cysteine Endopeptidases / genetics*
  • Cysteine Endopeptidases / isolation & purification
  • Cysteine Endopeptidases / metabolism*
  • Cysteine Proteinase Inhibitors / pharmacology
  • Dipeptides / metabolism
  • Gene Expression
  • Gene Library
  • Gnathostoma / enzymology*
  • Gnathostoma / genetics
  • Humans
  • Molecular Sequence Data
  • Molecular Weight
  • Open Reading Frames
  • Phylogeny
  • Pichia / genetics
  • Protein Sorting Signals
  • Protein Structure, Tertiary
  • Sequence Alignment
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Substrate Specificity


  • Coumarins
  • Cysteine Proteinase Inhibitors
  • Dipeptides
  • Protein Sorting Signals
  • benzyloxycarbonyl-phenylalanylarginine-4-methylcoumaryl-7-amide
  • benzyloxycarbonylarginyl-arginine 4-methylcoumarin-7-ylamide
  • Cathepsins
  • Cysteine Endopeptidases
  • CTSL protein, human
  • Cathepsin L
  • Ctsl protein, mouse

Associated data

  • GENBANK/EU327528