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Altered synthesis of cartilage-specific proteoglycans by mutant human cartilage oligomeric matrix protein

Authors
 Yoon Hae Kwak  ;  Jae Young Roh  ;  Ki Seok Lee  ;  Hui Wan Park  ;  Hyun Woo Kim 
Citation
 CLINICS IN ORTHOPEDIC SURGERY, Vol.1(4) : 181-187, 2009 
Journal Title
CLINICS IN ORTHOPEDIC SURGERY
ISSN
 2005-291X 
Issue Date
2009
MeSH
Aggrecans/analysis ; Aggrecans/biosynthesis* ; Animals ; CartilageOligomericMatrixProtein ; Cells, Cultured ; Chondrocytes/metabolism* ; Chondrosarcoma/metabolism ; Collagen Type II/biosynthesis* ; ExtracellularMatrix/metabolism* ; ExtracellularMatrixProteins/genetics* ; Glycoproteins/genetics* ; Humans ; Matrilin Proteins ; Mutation ; Rats ; Transfection
Keywords
Extracellularmatrix ; MutantCOMP ; Pseudoachondroplasia
Abstract
BACKGROUND: The mechanism by which mutant cartilage oligomeric matrix protein (COMP) induces a pseudoachondroplasia phenotype remains unknown, and the reason why a mutation of a minor protein of the growth plate cartilage causes total disruption of endochondral bone formation has not yet been determined. The current study was performed to investigate the effects of mutated COMP on the synthesis of the cartilage-specific major matrix proteins of Swarm rat chondrosarcoma chondrocytes.

METHODS: The Swarm rat chondrosarcoma chondrocytes transfected with a chimeric construct, which consisted of a mutant gene of human COMP and an amino acid FLAG tag sequence, were cultured in agarose gel. Formation of extracellular proteoglycan and type-II collagen by the cells was evaluated by immunohistochemical staining and measuring the (35)S-sulfate incorporation.

RESULTS: No difference was observed for the detection of type-II collagen among the cell lines expressing mutant COMP and the control cell lines. Histochemical staining of sulfated proteoglycans with safranin-O showed that lesser amounts of proteoglycans were incorporated into the extracellular matrix of the chondrocytes transfected with the mutant gene. (35)S-sulfate incorporation into the cell/matrix fractions demonstrated markedly lower radiolabel incorporation, as compared to that of the control cells.

CONCLUSIONS: Mutation of COMP has an important impact on the processing of proteoglycans, rather than type-II collagen, in the three-dimensional culture of Swarm rat chondrosarcoma chondrocytes
Files in This Item:
T200904862.pdf Download
DOI
10.4055/cios.2009.1.4.181
Appears in Collections:
1. College of Medicine (의과대학) > Dept. of Orthopedic Surgery (정형외과학교실) > 1. Journal Papers
Yonsei Authors
Kim, Hyun Woo(김현우) ORCID logo https://orcid.org/0000-0001-8576-1877
Park, Hui Wan(박희완)
Lee, Ki Seok(이기석)
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/105530
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