RRC ID |
41360
|
著者 |
Ito H, Akiyama H, Iguchi H, Iyama K, Miyamoto M, Ohsawa K, Nakamura T.
|
タイトル |
Molecular cloning and biological activity of a novel lysyl oxidase-related gene expressed in cartilage.
|
ジャーナル |
J Biol Chem
|
Abstract |
We cloned a cDNA encoding a novel lysyl oxidase-related protein, named LOXC, by suppression subtractive hybridization between differentiated and calcified ATDC5 cells, a clonal mouse chondrogenic EC cell line. The deduced amino acid sequence of mouse LOXC consists of 757 amino acids and shows 50% identity with that of mouse lysyl oxidase. Northern blot analysis showed a distinct hybridization band of 5.4 kilobases, and Western blot analysis showed an immunoreactive band at 82 kilodaltons. Expression of LOXC mRNA was detected in osteoblastic MC3T3-E1 cells and embryonic fibroblast C3H10T1/2 cells, whereas none of NIH3T3 fibroblasts and myoblastic C2C12 cells expressed LOXC mRNA in vitro. Moreover, the LOXC mRNA and protein levels dramatically increased throughout a process of chondrogenic differentiation in ATDC5 cells. In vivo, LOXC gene expression was localized in hypertrophic and calcified chondrocytes of growth plates in adult mice. The conditioned media of COS-7 cells transfected with the full-length LOXC cDNA showed the lysyl oxidase activity in both type I and type II collagens derived from chick embryos, and these activities of LOXC were inhibited by beta-aminopropionitrile, a specific inhibitor of lysyl oxidase. Our data indicate that LOXC is expressed in cartilage in vivo and modulates the formation of a collagenous extracellular matrix.
|
巻・号 |
276(26)
|
ページ |
24023-9
|
公開日 |
2001-6-29
|
DOI |
10.1074/jbc.M100861200
|
PII |
S0021-9258(20)78408-5
|
PMID |
11292829
|
MeSH |
Amino Acid Oxidoreductases*
Amino Acid Sequence
Animals
Animals, Newborn
Base Sequence
Blotting, Western
COS Cells
Cartilage / enzymology*
Cell Line
Cloning, Molecular
Growth Plate / enzymology
Mice
Molecular Sequence Data
Protein-Lysine 6-Oxidase / genetics*
Protein-Lysine 6-Oxidase / metabolism*
RNA, Messenger / biosynthesis
Sequence Homology, Amino Acid
Tissue Distribution
Transcription, Genetic
Transfection
|
IF |
4.238
|
引用数 |
51
|
WOS 分野
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
リソース情報 |
ヒト・動物細胞 |
NIH3T3
C2C12(RCB0987)
10T1/2(RCB0247)
MC3T3-E1(RCB1126) |