The substrate specificity of Uca pugilator collagenolytic serine protease 1 correlates with the bovine type I collagen cleavage sites

J Biol Chem. 1994 Jul 29;269(30):19565-72.


Affinity-based purification and characterization of the collagenolytic serine protease 1 from Uca pugilator (fiddler crab) hepatopancreas shows that the enzyme cleaves the native bovine alpha 1(I) collagen chain carboxyl-terminal to Gln and Arg residues adjacent to the metallocollagenase site. Cleavage carboxyl-terminal to Leu residues is observed in the alpha 2(I) chain and at a secondary site in alpha 1(I). These sites correlate with the preferences observed toward p-nitroanilide substrates varying at the P1 position, for which the specificity (kcat/Km) is Arg > Leu, Phe, Lys > Gln > Ala. Furthermore, collagen cleavage after Gln was found exclusively between two Gln-Arg bonds. The P'1-P'3 specificity of collagenase, as determined by nucleophile acyl transfer, indicated a strong preference for Arg in the P'1 position. Crab collagenase cleaves peptide bonds adjacent to Leu and Gln at the P1 position more efficiently than trypsin, chymotrypsin, or elastase. Moreover, the efficiency of collagenase toward P1-Arg substrates is equivalent to that of trypsin. Crystals of crab collagenase have been grown complexed with the protein inhibitor ecotin. These crystals diffract to better than 2.8 A resolution and belong to the space group P3(2)21 with unit cell dimensions of a = b = 89.0 A, c = 291.7 A.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Brachyura / enzymology*
  • Chymotrypsin / genetics
  • Collagen / metabolism*
  • Crystallization
  • Escherichia coli Proteins*
  • Esterases / metabolism
  • Liver / enzymology*
  • Molecular Sequence Data
  • Pancreas / enzymology*
  • Pancreatic Elastase / genetics
  • Periplasmic Proteins*
  • Protein Binding
  • Sequence Analysis
  • Sequence Homology, Amino Acid
  • Serine Endopeptidases / genetics
  • Serine Endopeptidases / isolation & purification
  • Serine Endopeptidases / metabolism*
  • Substrate Specificity
  • Trypsin / genetics
  • Trypsin Inhibitors / metabolism


  • Bacterial Proteins
  • Eco protein, E coli
  • Escherichia coli Proteins
  • Periplasmic Proteins
  • Trypsin Inhibitors
  • Collagen
  • Esterases
  • Serine Endopeptidases
  • collagenolytic protease I, crab
  • Chymotrypsin
  • Pancreatic Elastase
  • Trypsin