RRC ID 35669
著者 Miyamori H, Takino T, Kobayashi Y, Tokai H, Itoh Y, Seiki M, Sato H.
タイトル Claudin promotes activation of pro-matrix metalloproteinase-2 mediated by membrane-type matrix metalloproteinases.
ジャーナル J Biol Chem
Abstract Genes associated with regulation of membrane-type matrix metalloproteinase-1 (MT1-MMP)-mediated pro-MMP-2 processing were screened in 293T cells by a newly developed expression cloning method. One of the gene products, which promoted processing of pro-MMP-2 by MT1-MMP was claudin-5, a major component of endothelial tight junctions. Expression of claudin-5 not only replaced TIMP-2 in pro-MMP-2 activation by MT1-MMP but also promoted activation of pro-MMP-2 mediated by all MT-MMPs and MT1-MMP mutants lacking the transmembrane domain (DeltaMT1-MMP). A carboxyl-terminal deletion mutant of pro-MMP-2 (proDeltaMMP-2) was processed to an intermediate form by MT1-MMP in 293T cells and was further converted to an activated form by introduction of claudin-5. In contrast to the stimulatory effect of TIMP-2 on pro-MMP-2 activation by MT1-MMP, activation of pro-MMP-2 by DeltaMT1-MMP in the presence of claudin-5 and proDeltaMMP-2 processing by MT1-MMP were both inversely repressed by expression of exogenous TIMP-2. These results suggest that TIMP-2 is not involved in cluadin-5-induced pro-MMP-2 activation by MT-MMPs. Stimulation of MT-MMP-mediated pro-MMP-2 activation was also observed with other claudin family members, claudin-1, claudin-2, and claudin-3. Amino acid substitutions or deletions in ectodomain of claudin-1 abolished stimulatory effect. Direct interaction of claudin-1 with MT1-MMP and MMP-2 was demonstrated by immunoprecipitation analysis. MT1-MMP was co-localized with claudin-1 not only at cell-cell borders, but also at other parts of the cells. TIMP-2 enhanced cell surface localization of MMP-2 mediated by MT1-MMP, and claudin-1 also stimulated it. These results suggest that claudin recruits all MT-MMPs and pro-MMP-2 on the cell surface to achieve elevated focal concentrations and, consequently, enhances activation of pro-MMP-2.
巻・号 276(30)
ページ 28204-11
公開日 2001-7-27
DOI 10.1074/jbc.M103083200
PII S0021-9258(19)31627-8
PMID 11382769
MeSH Animals COS Cells Cell Line Cell Membrane / enzymology* Claudin-1 Claudin-3 Claudin-5 Claudins Cloning, Molecular DNA, Complementary / metabolism Enzyme Activation Enzyme-Linked Immunosorbent Assay Gene Deletion Gene Library Green Fluorescent Proteins Humans Luminescent Proteins / metabolism Matrix Metalloproteinase 14 Matrix Metalloproteinase 2 / metabolism* Matrix Metalloproteinases, Membrane-Associated Membrane Proteins / chemistry* Membrane Proteins / metabolism Membrane Proteins / physiology* Metalloendopeptidases / metabolism* Microscopy, Fluorescence Plasmids / metabolism Precipitin Tests Protein Binding Protein Structure, Tertiary Recombinant Fusion Proteins / metabolism Tight Junctions Tissue Inhibitor of Metalloproteinase-2 / metabolism Transfection
IF 4.238
引用数 182
WOS 分野 BIOCHEMISTRY & MOLECULAR BIOLOGY
リソース情報
遺伝子材料 pEAK-Claudin-5 (RDB02846)