title: Further characterization of interaction between bone sialoprotein (BSP) and collagen.
abstract: Bone sialoprotein (BSP) has an affinity to collagen fibrils [25]. A role of carbohydrate chains in the affinity was examined by removing sialic acids of BSP. Neuraminidase treatment of the BSP increased the binding to collagen. Binding sites of BSP on collagen were examined by biochemical and electron-microscopic methods. Purified bovine BSP was labeled with biotin. Collagen alpha chains or CNBr peptides were separated by electrophoresis and transfered to nitrocellulose membranes. The membranes were incubated with the biotin-labeled BSP, and the bound BSP was visualized with avidin conjugated with alkaline phosphatase. The labeled BSP was preferentially bound to the alpha 2 chain, and peptides derived from alpha 2 chain. In another experiment, the labeled BSP was incubated with reconstituted native collagen fibrils. The mixture was put on a copper grid, reacted with avidin conjugated with gold particles, and observed with an electron microscope. The gold particles were seen mainly within hole zones of the fibrils. BSP bound to the alpha 2 chain within the hole zones may regulate the onset of calcification at hole zones and the cell binding to collagen fibrils.
authors.affiliation: Department of Biochemistry, Hokkaido University School of Dentistry, Sapporo, Japan.
authors.forename: R
authors.initials: R
authors.lastname: Fujisawa
authors.forename: Y
authors.initials: Y
authors.lastname: Nodasaka
authors.forename: Y
authors.initials: Y
authors.lastname: Kuboki
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D003094:Collagen / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D066298:In Vitro Techniques; D058574:Integrin-Binding Sialoprotein; D008854:Microscopy, Electron; D009439:Neuraminidase; D011485:Protein Binding; D012794:Sialic Acids / Q000737:chemistry; D012795:Sialoglycoproteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D058574:Integrin-Binding Sialoprotein; D012794:Sialic Acids; D012795:Sialoglycoproteins; D003094:Collagen; D009439:Neuraminidase