Induction of matrix metalloproteinase activation cascades based on membrane-type 1 matrix metalloproteinase: associated activation of gelatinase A, gelatinase B and collagenase 3

• Published in: Biochemical journal Vol. 331 ( Pt 2); no. 2; pp. 453 - 458
• Main Authors: Cowell, S, Knäuper, V, Stewart, M L, D'Ortho, M P, Stanton, H, Hembry, R M, López-Otín, C, Reynolds, J J, Murphy, G
• Format: Journal Article
• Language: English
• Published: England 15.04.1998
• Subjects: Blotting, Western; Cell Membrane - enzymology; Chondrosarcoma - enzymology; Collagenases - biosynthesis; Collagenases - metabolism; Concanavalin A - pharmacology; Culture Media, Conditioned; Enzyme Activation; Enzyme Induction - drug effects; Fibrinolysin - physiology; Gelatinases - metabolism; Humans; Interleukin-1 - pharmacology; Matrix Metalloproteinase 13; Matrix Metalloproteinase 2; Matrix Metalloproteinase 9; Metalloendopeptidases - metabolism; Protease Inhibitors - pharmacology; Tetradecanoylphorbol Acetate - pharmacology; Tissue Inhibitor of Metalloproteinase-1 - metabolism; Tissue Inhibitor of Metalloproteinase-1 - pharmacology; Tissue Inhibitor of Metalloproteinase-2 - metabolism; Tissue Inhibitor of Metalloproteinase-2 - pharmacology; Tissue Inhibitor of Metalloproteinase-3 - pharmacology; Tumor Cells, Cultured
• ISSN: 0264-6021; 1470-8728
• DOI: 10.1042/bj3310453

SW1353 chondrosarcoma cells cultured in the presence of interleukin-1, concanavalin A or PMA secreted procollagenase 3 (matrix metalloproteinase-13). The enzyme was detected in the culture medium by Western blotting using a specific polyclonal antibody raised against recombinant human procollagenase 3. Oncostatin M enhanced the interleukin-1-induced production of procollagenase 3, whereas interleukin-4 decreased procollagenase 3 synthesis. The enzyme was latent except when the cells had been treated with concanavalin A, when a processed form of 48 kDa, which corresponds to the active form, was found in the culture medium and collagenolytic activity was detected by degradation of 14C-labelled type I collagen. The concanavalin A-induced activation of procollagenase 3 coincided with the processing of progelatinase A (matrix metalloproteinase-2) by the cells, as measured by gelatin zymography. In addition, progelatinase B (matrix metalloproteinase-9) was activated when gelatinase A and collagenase 3 were in their active forms. Concanavalin A treatment of SW1353 cells increased the amount of membrane-type-1 matrix metalloproteinase protein in the cell membranes, suggesting that this membrane-bound enzyme participates in an activation cascade involving collagenase 3 and the gelatinases. This cascade was effectively inhibited by tissue inhibitors of metalloproteinases-2 and -3. Tissue inhibitor of metalloproteinases-1, which is a much weaker inhibitor of membrane-type 1 matrix metalloproteinase than tissue inhibitors of metalloproteinases-2 and -3 [Will, Atkinson, Butler, Smith and Murphy (1996) J. Biol. Chem. 271, 17119-17123], was a weaker inhibitor of the activation cascade.