Secretion of metalloproteinases by stimulated capillary endothelial cells. II. Expression of collagenase and stromelysin activities is regulated by endogenous inhibitors.
Rabbit brain capillary endothelial cells treated with 12-O-tetradecanoylphorbol-13-acetate produce the metalloproteinases, procollagenase and prostromelysin, as up to 20% of their total secreted protein. However, little or no catalytic activity of these enzymes can be found after treatment with either trypsin or an organomercurial agent, which are able to activate the proenzymes in the medium from stimulated rabbit fibroblasts. We now have shown that enzyme activities of procollagenase and prostromelysin are revealed after conditioned medium is analyzed by gel filtration chromatography or by electrophoresis on sodium dodecyl sulfate-substrate gels. In both systems, the metalloproteinases were separated from metalloproteinase inhibitors. The major inhibitor of Mr = 30,000 from capillary endothelial cells was immunologically identical with the rabbit tissue inhibitor of metalloproteinases. Two additional inhibitors of metalloproteinases at Mr = 22,000 and 19,000 were also observed. Inhibitors were present in the conditioned medium from rabbit fibroblasts in much lower quantities and were also qualitatively different. When gel filtration chromatography was used to remove the tissue inhibitor of metalloproteinases from medium conditioned by stimulated capillary endothelial cells, both activatable procollagenase and prostromelysin were readily demonstrable. These data suggest that endogenous inhibitors regulate the expression of metalloproteinases secreted by endothelial cells.
D000818:Animals; D002196:Capillaries / Q000201:enzymology; D002560:Cerebrovascular Circulation; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004727:Endothelium / Q000201:enzymology; D006706:Homeostasis; D019278:Matrix Metalloproteinase 3; D003012:Microbial Collagenase / Q000037:antagonists & inhibitors*; D008970:Molecular Weight; D011480:Protease Inhibitors; D011817:Rabbits; D013997:Time Factors
D011480:Protease Inhibitors; D019278:Matrix Metalloproteinase 3; D003012:Microbial Collagenase