title: Mode of hydrolysis of collagen-like peptides by class I and class II Clostridium histolyticum collagenases: evidence for both endopeptidase and tripeptidylcarboxypeptidase activities.
abstract: The action of three class I (beta, gamma, and eta) and three class II (delta, epsilon, and zeta) collagenases from Clostridium histolyticum on two series of peptides with collagen-like sequences has been examined. The peptides in the first series all contain 4-nitrophenylalanyl-Gly-Pro-Ala in subsites P1 through P3', but each is successively lengthened in the N-terminal direction by addition of an appropriate residue until subsite P5 is occupied. The second group of peptides all have cinnamoyl-Leu in subsites P2 and P1, respectively, but each is successively lengthened in the C-terminal direction by partial additions of the Gly-Pro-Leu triplet until subsite P6' is occupied. N-Terminal elongation causes the kcat/KM values to rise markedly and to level off after occupancy of subsite P6 for the class I enzymes and subsite P3 for the class II enzymes. C-Terminal elongation produces the best substrates for both classes of enzymes when subsites P3' or P4' are occupied by amino acids with free carboxyl groups. The kcat/KM values for the hydrolysis of both Leu-Gly bonds of cinnamoyl-Leu-Gly-Pro-Leu-Gly-Pro-Leu have been measured for both classes of enzymes. Both rates are large, but both classes preferentially hydrolyze the Leu-Gly bond of the C-terminal triplet. Thus, both classes of enzymes exhibit both endopeptidase and tripeptidylcarboxypeptidase activities.
mesh_terms: D000595:Amino Acid Sequence; D002268:Carboxypeptidases; D003013:Clostridium / Q000201:enzymology*; D003094:Collagen / Q000378:metabolism*; D010450:Endopeptidases; D006868:Hydrolysis; D007527:Isoenzymes / Q000378:metabolism*; D003012:Microbial Collagenase / Q000378:metabolism*; D009842:Oligopeptides; D013379:Substrate Specificity
chemical_list: D007527:Isoenzymes; D009842:Oligopeptides; D003094:Collagen; D002268:Carboxypeptidases; D010450:Endopeptidases; D003012:Microbial Collagenase