The activation of human skin fibroblast procollagenase. Sequence identification of the major conversion products.
Human skin collagenase is secreted by cultured fibroblasts in a proenzyme form and can be activated to a catalytically competent enzyme by a number of processes. All modes of activation studied lead to conversion of the proenzyme to a stable 42-kDa active enzyme, concomitant with removal of an 81-amino acid peptide from the amino-terminal end of the molecule. The sequence of events leading to the formation of this enzyme form has been determined by analyzing the primary structure of the conversion intermediates. Trypsin-induced activation of procollagenase occurs as a result of the initial cleavage of the peptide bond between Arg-55 and Asn-56, generating a major intermediate of 46 kDa. Treatment of the proenzyme with organomercurials, which have no intrinsic ability to cleave peptide bonds, initially results in activation of the enzyme without loss of molecular weight. This is followed by conversion to two lower molecular weight species of 44 and 42 kDa, the latter corresponding to the stable active enzyme form. The final cleavage producing this form of collagenase is not restricted to a single polypeptide bond but can occur on the amino-terminal side of any one of three contiguous hydrophobic residues, Phe-100, Val-101, Leu-102. The data suggest that both trypsin and organomercurials activate procollagenase by initiating an intramolecular autoproteolytic reaction resulting in the formation of a stable 42-kDa active enzyme species.
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D017364:Collagenases; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D003012:Microbial Collagenase / Q000378:metabolism*; D008970:Molecular Weight; D009941:Organomercury Compounds / Q000494:pharmacology; D012867:Skin / Q000201:enzymology*; D014357:Trypsin / Q000378:metabolism
D004792:Enzyme Precursors; D009941:Organomercury Compounds; D014357:Trypsin; D017364:Collagenases; C022440:procollagenase; D003012:Microbial Collagenase