WARP interacts with collagen VI-containing microfibrils in the pericellular matrix of human chondrocytes

PLoS One. 2012;7(12):e52793. doi: 10.1371/journal.pone.0052793. Epub 2012 Dec 26.


Collagen VI and WARP are extracellular structural macromolecules present in cartilage and associated with BM suprastructures in non-skeletal tissues. We have previously shown that in WARP-deficient mice, collagen VI is specifically reduced in regions of the peripheral nerve ECM where WARP is expressed, suggesting that both macromolecules are part of the same suprastructure. The object of this study was to conduct a detailed analysis of WARP-collagen VI interactions in vitro in cartilage, a tissue rich in WARP and collagen VI. Immunohistochemical analysis of mouse and human articular cartilage showed that WARP and collagen VI co-localize in the pericellular matrix of superficial zone articular chondrocytes. EM analysis on extracts of human articular cartilage showed that WARP associates closely with collagen VI-containing suprastructures. Additional evidence of an interaction is provided by immunogold EM and immunoblot analysis showing that WARP was present in collagen VI-containing networks isolated from cartilage. Further characterization were done by solid phase binding studies and reconstitution experiments using purified recombinant WARP and isolated collagen VI. Collagen VI binds to WARP with an apparent K(d) of approximately 22 nM and the binding site(s) for WARP resides within the triple helical domain since WARP binds to both intact collagen VI tetramers and pepsinized collagen VI. Together, these data confirm and extend our previous findings by demonstrating that WARP and collagen VI form high affinity associations in vivo in cartilage. We conclude that WARP is ideally placed to function as an adapter protein in the cartilage pericellular matrix.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cartilage, Articular / cytology
  • Cartilage, Articular / metabolism
  • Cartilage, Articular / ultrastructure
  • Cattle
  • Chondrocytes / metabolism*
  • Collagen Type VI / chemistry
  • Collagen Type VI / metabolism*
  • Collagen Type VI / ultrastructure
  • Extracellular Matrix / metabolism
  • Extracellular Matrix / ultrastructure
  • Extracellular Matrix Proteins / chemistry
  • Extracellular Matrix Proteins / metabolism*
  • Heparan Sulfate Proteoglycans / metabolism
  • Humans
  • Immunoprecipitation
  • Mice
  • Microfibrils / chemistry
  • Microfibrils / metabolism*
  • Microfibrils / ultrastructure
  • Protein Binding


  • Collagen Type VI
  • Extracellular Matrix Proteins
  • Heparan Sulfate Proteoglycans
  • VWA1 protein, human
  • perlecan