title: Metalloproteinase activity secreted by fibrogenic cells in the processing of prolysyl oxidase. Potential role of procollagen C-proteinase.
abstract: Lysyl oxidase is secreted from fibrogenic cells as a 50-kDa proenzyme that is proteolytically processed to the mature enzyme in the extracellular space. To characterize the secreted proteinase activity, a truncated, recombinant form of lysyl oxidase was prepared as a proteinase substrate containing the sequence of the propeptide cleavage region. The processing proteinase activity secreted by cultured fibrogenic cells resists inhibitors of serine or aspartyl proteinases as well as tissue inhibitor of matrix metalloproteinases-2 (MMP-2) but is completely inhibited by metal ion chelators. Known metalloproteinases were tested for their activity toward this substrate. Carboxyl-terminal procollagen proteinase (C-proteinase), MMP-2, and conditioned fibrogenic cell culture medium cleave the lysyl oxidase substrate to the size of the mature enzyme. The NH2-terminal sequence generated by arterial smooth muscle conditioned medium and the C-proteinase but not by MMP-2, i.e. Asp-Asp-Pro-Tyr, was identical to that previously identified in mature lysyl oxidase isolated from connective tissue. The C-proteinase activity against the model substrate was inhibited by a synthetic oligopeptide mimic of the cleavage sequence (Ac-Met-Val-Gly-Asp-Asp-Pro-Tyr-Asn-amide), whereas this peptide also inhibited the generation of lysyl oxidase activity in the medium of fetal rat lung fibroblasts in culture. In toto, these results identify a secreted metalloproteinase activity participating in the activation of prolysyl oxidase, identify inhibitors of the processing activity, and implicate procollagen C-proteinase in this role.
authors.affiliation: Department of Biochemistry, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
authors.forename: M V
authors.initials: MV
authors.lastname: Panchenko
authors.forename: W G
authors.initials: WG
authors.lastname: Stetler-Stevenson
authors.forename: O V
authors.initials: OV
authors.lastname: Trubetskoy
authors.forename: S N
authors.initials: SN
authors.lastname: Gacheru
authors.forename: H M
authors.initials: HM
authors.lastname: Kagan
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D055395:Bone Morphogenetic Protein 1; D019485:Bone Morphogenetic Proteins; D002478:Cells, Cultured; D003238:Connective Tissue / Q000201:enzymology; D006801:Humans; D006868:Hydrolysis; D008666:Metalloendopeptidases / Q000378:metabolism*; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000201:enzymology; D009838:Oligodeoxyribonucleotides; D011498:Protein Precursors / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D008249:Protein-Lysine 6-Oxidase / Q000378:metabolism*; D051381:Rats; D017386:Sequence Homology, Amino Acid
chemical_list: D019485:Bone Morphogenetic Proteins; D009838:Oligodeoxyribonucleotides; D011498:Protein Precursors; D008249:Protein-Lysine 6-Oxidase; D008666:Metalloendopeptidases; C525705:BMP1 protein, human; C525708:Bmp1 protein, rat; D055395:Bone Morphogenetic Protein 1