Characterization of recombinant amino-terminal NC4 domain of human collagen IX - Interaction with glycosaminoglycans and cartilage oligomeric matrix protein

Julkaisun otsikon käännös: Ihmisen kollageeni IX:n aminoterminaalisen rekombinantti NC4-domeinin karakterisointi - vuorovaikutus proteoglykaanien ja COMP-proteiinin kanssa

T Pihlajamaa, H Lankinen, J Ylostalo, L Valmu, J Jaalinoja, F Zaucke, L Spitznagel, S Gosling, A Puustinen, M Morgelin, J Peranen, P Maurer, L Ala-Kokko, I Kilpelainen

    Tutkimustuotos: ArtikkelijulkaisuArtikkeliTieteellinenvertaisarvioitu

    Abstrakti

    The N-terminal NC4 domain of collagen IX is a globular structure projecting away from the surface of the cartilage collagen fibril. Several interactions have been suggested for this domain, reflecting its location and its characteristic high isoelectric point. In an attempt to characterize the NC4 domain in more detail, we set up a prokaryotic expression system to produce the domain. The purified 27.5-kDa product was analyzed for its glycosaminoglycan-binding potential by surface plasmon resonance and solid-state assays. The results show that the NC4 domain of collagen IX specifically binds heparin with a K(d) of 0.6 microm, and the full-length recombinant collagen IX has an even stronger interaction with heparin, with an apparent K(d) of 3.6 nm. The heparin-binding site of the NC4 domain was located in the extreme N terminus, containing a heparin-binding consensus sequence, whereas electron microscopy suggested the presence of at least three additional heparin-binding sites on full-length collagen IX. The NC4 domain was also shown to bind cartilage oligomeric matrix protein. This interaction and the association of cartilage oligomeric matrix protein with other regions of collagen IX were found to be heparin-competitive. Circular dichroism analyses of the NC4 domain indicated the presence of stabilizing disulfide bonds and a thermal denaturation point of about 80 degrees C. The pattern of disulfide bond formation within the NC4 domain was identified by tryptic peptide mass mapping of the NC4 in native and reduced states. A similar pattern was demonstrated for the NC4 domain of full-length recombinant collagen IX.
    Julkaisun otsikon käännösIhmisen kollageeni IX:n aminoterminaalisen rekombinantti NC4-domeinin karakterisointi - vuorovaikutus proteoglykaanien ja COMP-proteiinin kanssa
    Alkuperäiskielienglanti
    LehtiJournal of Biological Chemistry
    Vuosikerta279
    Numero23
    Sivut24265-24273
    Sivumäärä9
    ISSN0021-9258
    DOI - pysyväislinkit
    TilaJulkaistu - 4 kesäkuuta 2004
    OKM-julkaisutyyppiA1 Alkuperäisartikkeli tieteellisessä aikakauslehdessä, vertaisarvioitu

    Tieteenalat

    • 118 Biotieteet
    • biokemia
    • collagen
    • cartilage
    • glycosaminoglycan
    • COMP

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