title: Dentin matrix protein 1 immobilized on type I collagen fibrils facilitates apatite deposition in vitro.
abstract: During bone and dentin mineralization, the crystal nucleation and growth processes are considered to be matrix regulated. Osteoblasts and odontoblasts synthesize a polymeric collagenous matrix, which forms a template for apatite initiation and elongation. Coordinated and controlled reaction between type I collagen and bone/dentin-specific noncollagenous proteins are necessary for well defined biogenic crystal formation. However, the process by which collagen surfaces become mineralized is not understood. Dentin matrix protein 1 (DMP1) is an acidic noncollagenous protein expressed during the initial stages of mineralized matrix formation in bone and dentin. Here we show that DMP1 bound specifically to type I collagen, with the binding region located at the N-telopeptide region of type I collagen. Peptide mapping identified two acidic clusters in DMP1 responsible for interacting with type I collagen. The collagen binding property of these domains was further confirmed by site-directed mutagenesis. Transmission electron microscopy analyses have localized DMP1 in the gap region of the collagen fibrils. Fibrillogenesis assays further demonstrated that DMP1 accelerated the assembly of the collagen fibrils in vitro and also increased the diameter of the reconstituted collagen fibrils. In vitro mineralization studies in the presence of calcium and phosphate ions demonstrated apatite deposition only at the collagen-bound DMP1 sites. Thus specific binding of DMP1 and possibly other noncollagenous proteins on the collagen fibril might be a key step in collagen matrix organization and mineralization.
authors.affiliation: Department of Oral Biology, University of Illinois at Chicago, Chicago, Illinois 60612, USA.
authors.forename: Gen
authors.initials: G
authors.lastname: He
authors.forename: Anne
authors.initials: A
authors.lastname: George
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001031:Apatites / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D024042:Collagen Type I / Q000737:chemistry / Q000378:metabolism*; D017931:DNA Primers; D016326:Extracellular Matrix Proteins; D066298:In Vitro Techniques; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010750:Phosphoproteins; D011485:Protein Binding; D051381:Rats; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*
chemical_list: D001031:Apatites; D024042:Collagen Type I; D017931:DNA Primers; C081635:Dmp1 protein, rat; D016326:Extracellular Matrix Proteins; D010750:Phosphoproteins; D014157:Transcription Factors