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Research article2010Peer reviewedOpen access

The Streptococcal Collagen-binding Protein CNE Specifically Interferes with alpha(V)beta(3)-mediated Cellular Interactions with Triple Helical Collagen

van Wieringen Tijs, Kalamajski Sebastian, Lidén Åsa, Bihan Dominique, Guss Bengt, Heinegård Dick, Farndale Richard W, Rubin Kristofer

Abstract

Collagen fibers expose distinct domains allowing for specific interactions with other extracellular matrix proteins and cells. To investigate putative collagen domains that govern integrin alpha(V)beta(3)-mediated cellular interactions with native collagen fibers we took advantage of the streptococcal protein CNE that bound native fibrillar collagens. CNE specifically inhibited alpha(V)beta(3)-dependent cell-mediated collagen gel contraction, PDGF BB-induced and alpha(V)beta(3)-mediated adhesion of cells, and binding of fibronectin to native collagen. Using a Toolkit composed of overlapping, 27-residue triple helical segments of collagen type II, two CNE-binding sites present in peptides II-1 and II-44 were identified. These peptides lack the major binding site for collagen-binding beta(1) integrins, defined by the peptide GFOGER. Peptide II-44 corresponds to a region of collagen known to bind collagenases, discoidin domain receptor 2, SPARC (osteonectin), and fibronectin. In addition to binding fibronectin, peptide II-44 but not II-1 inhibited alpha(V)beta(3)-mediated collagen gel contraction and, when immobilized on plastic, supported adhesion of cells. Reduction of fibronectin expression by siRNA reduced PDGF BB-induced alpha(V)beta(3)-mediated contraction. Reconstitution of collagen types I and II gels in the presence of CNE reduced collagen fibril diameters and fibril melting temperatures. Our data indicate that contraction proceeded through an indirect mechanism involving binding of cell-produced fibronectin to the collagen fibers. Furthermore, our data show that cell-mediated collagen gel contraction does not directly depend on the process of fibril formation.

Published in

Journal of Biological Chemistry
2010, Volume: 285, number: 46, pages: 35803-35813

    UKÄ Subject classification

    Animal and Dairy Science
    Veterinary Science

    Publication identifier

    DOI: https://doi.org/10.1074/jbc.M110.146001

    Permanent link to this page (URI)

    https://res.slu.se/id/publ/31305