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Comparative Proteomic Analysis of Normal and Collagen IX Null Mouse Cartilage Reveals Altered Extracellular Matrix Composition and Novel Components of the Collagen IX Interactome

journal contribution
posted on 2023-05-17, 16:59 authored by Brachvogel, B, Zaucke, F, Dave, K, Norris, EL, Stermann, J, Dayakli, M, Koch, M, Gorman, JJ, Bateman, JF, Richard WilsonRichard Wilson
The cartilage extracellular matrix is essential for endochondral bone development and joint function. In addition to the major aggrecan/collagen II framework, the interacting complex of collagen IX, matrilin-3 and COMP is essential for cartilage matrix stability, as mutations in Col9a1, Col9a2, Col9a3, Comp and Matn3 genes cause multiple epiphyseal dysplasia, in which patients develop early onset osteoarthritis. In mice, collagen IX ablation results in severely disturbed growth plate organization, hypocellular regions and abnormal chondrocyte shape. This abnormal differentiation is likely to involve altered cell-matrix interactions but the mechanism is not known. To investigate the molecular basis of the collagen IX null phenotype we analysed global differences in protein abundance between wild-type and knockout femoral head cartilage by capillary HPLC tandem mass spectrometry. We identified 297 proteins in 3-day cartilage and 397 proteins in 21-day cartilage. Components that were differentially abundant between wild-type and collagen IX deficient cartilage included 15 extracellular matrix proteins. Collagen IX ablation was associated with dramatically reduced COMP and matrilin-3, consistent with known interactions. Matrilin-1, matrilin-4, epiphycan and thrombospondin-4 levels were reduced in collagen IX null cartilage, providing the first in vivo evidence for these proteins belonging to the collagen IX interactome. Thrombospondin-4 expression was reduced at the mRNA level, whereas matrilin-4 was verified as a novel collagen IX –binding protein. Furthermore, changes in TGF-induced protein ig-h3 and fibronectin abundance were found in the collagen IX knockout but not associated with COMP ablation, indicating specific involvement in the abnormal collagen IX null cartilage. In addition, the more widespread expression of collagen XII in the collagen IX deficient cartilage suggests an attempted compensatory response to the absence of collagen IX. Our differential proteomic analysis of cartilage is a novel approach to identify candidate matrix protein interactions in vivo, underpinning further analysis of mutant cartilage lacking other matrix components or harboring disease-causing mutations.

Funding

University of Tasmania

History

Publication title

Journal of Biological Chemistry

Volume

288

Issue

19

Pagination

13481-13492

ISSN

0021-9258

Publisher

AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Place of publication

9650 ROCKVILLE PIKE, BETHESDA, MD 20814-3996 USA

Rights statement

Copyright 2013 The American Society for Biochemistry and Molecular Biology, Inc.

Repository Status

  • Restricted

Socio-economic Objectives

Expanding knowledge in the health sciences

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