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) |