title: Activation of gelatinase A (72-kDa type IV collagenase) induced by monensin in normal human fibroblasts.
abstract: In monolayer culture, fibroblasts secrete all matrix metalloproteinases, including gelatinase A (72-kDa type IV collagenase), as inactive zymogens. Whereas limited proteolysis by plasmin or other matrix metalloproteinases (MMPs) can accomplish the extracellular activation of other proenzymes in this family, gelatinase A proenzyme is uniquely refractory to cleavage by such proteinases. Previously it has been shown that fibroblasts cultured in the presumably more physiologic culture milieu of a type I collagen lattice can be induced to secrete active gelatinase A. In monolayer culture, however, the plant lectin concanavalin A will induce gelatinase A activation. Here we show that in monolayer culture activation of gelatinase A by normal fibroblasts is also induced by the sodium ionophore monensin. The monensin response is dose-dependent, time-dependent, requires protein synthesis, and is specific to gelatinase A among the secreted matrix metalloproteinases. The activator appears to be associated with cell membranes and may be membrane-type matrix metalloproteinase 1(MT-MMP1). Both mRNA and immunodetectable protein of MT-MMP1 are increased with monensin treatment while message for the protein inhibitor of gelatinase A, TIMP-2, is unchanged. The monensin-induced signal transduction pathway leading to gelatinase activation in monolayer culture appears to be different from the integrin-mediated pathway operative in the collagen lattice system. The tyrosine kinase inhibitor genistein blocks monensin activation of gelatinase A in monolayer culture. In contrast, genistein has no effect on proenzyme activation in the collagen lattice. Likewise, the cyclooxygenase inhibitor indomethacin abrogates the monensin effect in monolayer culture and can be reversed by addition of exogenous prostaglandin E2 (PGE2). Neither indomethacin nor PGE2 affects activation of gelatinase A in the collagen lattice.
authors.affiliation: Division of Dermatology, Washington University School of Medicine, St. Louis, Missouri 63110, USA.
authors.forename: L
authors.initials: L
authors.lastname: Li
authors.forename: K
authors.initials: K
authors.lastname: Akers
authors.forename: A Z
authors.initials: AZ
authors.lastname: Eisen
authors.forename: J L
authors.initials: JL
authors.lastname: Seltzer
mesh_terms: D002478:Cells, Cultured; D017364:Collagenases / Q000032:analysis; D017077:Culture Media, Conditioned / Q000737:chemistry; D003513:Cycloheximide / Q000494:pharmacology; D016861:Cyclooxygenase Inhibitors / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004792:Enzyme Precursors / Q000378:metabolism; D005347:Fibroblasts / Q000187:drug effects* / Q000201:enzymology; D018093:Gelatinases / Q000378:metabolism*; D019833:Genistein; D006801:Humans; D007213:Indomethacin / Q000494:pharmacology; D016023:Integrins / Q000502:physiology; D017136:Ion Transport / Q000187:drug effects; D007476:Ionophores / Q000494:pharmacology*; D007529:Isoflavones / Q000494:pharmacology; D020778:Matrix Metalloproteinase 2; D019278:Matrix Metalloproteinase 3 / Q000032:analysis; D008666:Metalloendopeptidases / Q000378:metabolism*; D008985:Monensin / Q000494:pharmacology*; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D011506:Proteins / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D012867:Skin / Q000166:cytology; D012964:Sodium / Q000502:physiology*; D019716:Tissue Inhibitor of Metalloproteinase-2
chemical_list: D017077:Culture Media, Conditioned; D016861:Cyclooxygenase Inhibitors; D004791:Enzyme Inhibitors; D004792:Enzyme Precursors; D016023:Integrins; D007476:Ionophores; D007529:Isoflavones; D011500:Protein Synthesis Inhibitors; D011506:Proteins; D019716:Tissue Inhibitor of Metalloproteinase-2; D008985:Monensin; D003513:Cycloheximide; D012964:Sodium; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D017364:Collagenases; D018093:Gelatinases; D008666:Metalloendopeptidases; C069860:progelatinase; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D015232:Dinoprostone; D007213:Indomethacin