title: Direct production of an activated matrix metalloproteinase-9 (gelatinase B) from mammalian cells.
abstract: Matrix metalloproteinase-9 (MMP-9) is produced by the inactive proform and activated by a proteolytic process. However, it has not been reported to produce the active form directly from cells, which has hindered the research to elicit the physiological roles of this enzyme. In this study, we prepared mutant MMP-9 containing the furin-recognizing sequence in the prodomain and showed that the mutant MMP-9 was secreted as the active form directly from CHO-K1 cells and primary hepatocytes after the gene was transfected. The secreted MMP-9 showed proteolytic activity without further activation and degraded collagen IV in vitro. In addition, the transfection of the gene into the liver resulted in the efficient expression of active MMP-9 in the liver and the serum in vivo.
authors.affiliation: Department of Biomolecular Engineering, Tokyo Institute of Technology, 4259 Nagatsuda, Midori-ku, Yokohama 226-8501, Japan. ywatanab@bio.titech.ac.jp
authors.forename: Y
authors.initials: Y
authors.lastname: Watanabe
authors.forename: K
authors.initials: K
authors.lastname: Hirakawa
authors.forename: T
authors.initials: T
authors.lastname: Haruyama
authors.forename: T
authors.initials: T
authors.lastname: Akaike
mesh_terms: D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D004789:Enzyme Activation; D045683:Furin; D015870:Gene Expression; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D016296:Mutagenesis; D013381:Subtilisins / Q000378:metabolism*
chemical_list: D013381:Subtilisins; D045683:Furin; D020780:Matrix Metalloproteinase 9