title: Expression of three membrane-type matrix metalloproteinases (MT-MMPs) in rat vascular smooth muscle cells and characterization of MT3-MMPs with and without transmembrane domain.
abstract: Matrix metalloproteinases (MMPs) produced by rat smooth muscle cells (SMCs) were investigated. SMCs expressed three kinds of membrane-type MMP, MT1-MMP, MT2-MMP, and MT3-MMP, and the MT-MMP expression was stimulated by the presence of serum. MT3-MMP was characterized further by cloning its cDNA. A rat MT3-MMP cDNA encoding 607 amino acids and a cDNA for its transmembrane domainless variant MT3-MMP-del were cloned from a rat SMC cDNA library; a human MT3-MMP cDNA was cloned from a fetal brain cDNA library. Human brain MT3-MMP was similar but not identical to the previously reported human placenta MT3-MMP (94.4% homology). When the MT3-MMP cDNA was expressed in COS-7 cells, endogenous progelatinase A was processed to the mature form. The transfection of rat MT3-MMP-del efficiently converted progelatinase A to the intermediate form but not to the mature one, indicating that the transmembrane domain is important for the complete processing of progelatinase A to maturation. Both MT3-MMP-del and MT3-MMP hydrolyzed gelatin and casein, indicating their broad substrate specificity. Results of experiments with a synthetic MMP inhibitor suggested that MT3-MMP-del and MT3-MMP are rapidly degraded immediately after maturation. The present study suggests that multiple forms of MMPs including MT3-MMP are involved in the matrix remodeling of blood vessels.
authors.affiliation: Division of Cell Biology, Kihara Institute for Biological Research, Yokohama City University, 641-12 Maioka-cho, Totsuka-ku, Yokohama 244, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Shofuda
authors.forename: H
authors.initials: H
authors.lastname: Yasumitsu
authors.forename: A
authors.initials: A
authors.lastname: Nishihashi
authors.forename: K
authors.initials: K
authors.lastname: Miki
authors.forename: K
authors.initials: K
authors.lastname: Miyazaki
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D019556:COS Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007527:Isoenzymes / Q000235:genetics*; D053517:Matrix Metalloproteinase 15; D053513:Matrix Metalloproteinase 16; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000235:genetics*; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats
chemical_list: D018076:DNA, Complementary; D007527:Isoenzymes; C507098:MMP15 protein, human; C507102:MMP16 protein, human; D012333:RNA, Messenger; D053517:Matrix Metalloproteinase 15; D053513:Matrix Metalloproteinase 16; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases