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ID 50665
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Author
Hoshijima, Mitsuhiro
Yamashiro, Takashi
Takigawa, Masaharu Kaken ID publons researchmap
Abstract
To identify proteins that regulate CCN2 activity, we carried out GAL4-based yeast two-hybrid screening with a cDNA library derived from a chondrocytic cell line, HCS-2/8. CCN2/CTGF and CCN3/NOV polypeptides were picked up as CCN2-binding proteins, and CCN2CCN2 and CCN2CCN3 binding domains were identified. Direct binding between CCN2 and CCN3 was confirmed by coimmunoprecipitation in vitro and in vivo and surface plasmon resonance, and the calculated dissociation constants (Kd) were 1.17 x 10-9 m for CCN2 and CCN2, and 1.95 x 10-9 m for CCN2 and CCN3. Ectopically overexpressed green fluorescent proteinCCN2 and HaloCCN3 in COS7 cells colocalized, as determined by direct fluorescence analysis. We present evidence that CCN2CCN3 interactions modulated CCN2 activity such as enhancement of ACAN and col2a1 expression. Curiously, CCN2 enhanced, whereas CCN3 inhibited, the expression of aggrecan and col2a1 mRNA in HCS-2/8 cells, and combined treatment with CCN2 and CCN3 abolished the inhibitory effect of CCN3. These effects were neutralized with an antibody against the von Willebrand factor type C domain of CCN2 (11H3). This antibody diminished the binding between CCN2 and CCN2, but enhanced that between CCN3 and CCN2. Our results suggest that CCN2 could form homotypic and heterotypic dimers with CCN2 and CCN3, respectively. Strengthening the binding between CCN2 and CCN3 with the 11H3 antibody had an enhancing effect on aggrecan expression in chondrocytes, suggesting that CCN2 had an antagonizing effect by binding to CCN3.
Keywords
ACAN
CCN2
CTGF
CCN3
NOV
chondrocyte
dimerization
Published Date
2012-10
Publication Title
FEBS Journal
Volume
volume279
Issue
issue19
Publisher
Wiley-Blackwell
Start Page
3597
End Page
3584
ISSN
1742-464X
Content Type
Journal Article
Official Url
http://dx.doi.org/10.1111/j.1742-4658.2012.08717.x
Related Url
http://ousar.lib.okayama-u.ac.jp/metadata/50700
language
英語
File Version
author
Refereed
True
DOI
Web of Science KeyUT