Bone acidic glycoprotein-75 delineates the extracellular sites of future bone sialoprotein accumulation and apatite nucleation in osteoblastic cultures.
Addition of an organophosphate source to UMR osteoblastic cultures activates a mineralization program in which BSP localizes to extracellular matrix sites where hydroxyapatite crystals are subsequently nucleated. This study identifies for the first time novel extracellular spherical structures, termed biomineralization foci (BMF), containing bone acidic glycoprotein-75 (BAG-75), bone sialoprotein (BSP), and alkaline phosphatase that are the exclusive sites of initial nucleation of hydroxyapatite crystals in the UMR model. Importantly, in the absence of added phosphate, UMR cultures after reaching confluency contain two size populations of morphologically identifiable BMF precursors enriched in BAG-75 (15-25 and 150-250 microm in diameter). The shape and size of the smaller population are similar to structures assembled in vitro through self-association of purified BAG-75 protein. After organophosphate addition, BSP accumulates within these BAG-75-containing BMF precursors, with hydroxyapatite crystal nucleation occurring subsequently. In summary, BAG-75 is the earliest detectable biomarker that accurately predicts the extracellular sites of de novo biomineralization in UMR cultures. We hypothesize that BAG-75 may perform a key structural role in the assembly of BMF precursors and the recruitment of other proteins such as alkaline phosphatase and BSP. Furthermore, we propose a hypothetical mechanism in which BAG-75 and BSP function actively in nucleation of apatite within BMF.
Department of Biomedical Engineering and the Orthopaedic Research Center, Lerner Research Institute, The Cleveland Clinic and Foundation, Cleveland, Ohio 44195, USA.
Ronald J
RJ
Midura
Aimin
A
Wang
Dinah
D
Lovitch
Douglas
D
Law
Kimerly
K
Powell
Jeff P
JP
Gorski
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D001031:Apatites / Q000378:metabolism*; D015415:Biomarkers; D001842:Bone and Bones / Q000737:chemistry / Q000648:ultrastructure; D002113:Calcification, Physiologic; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000378:metabolism*; D006023:Glycoproteins / Q000032:analysis / Q000502:physiology*; D007150:Immunohistochemistry; D058574:Integrin-Binding Sialoprotein; D010006:Osteoblasts / Q000378:metabolism*; D051381:Rats; D012795:Sialoglycoproteins / Q000378:metabolism*
D001031:Apatites; D015415:Biomarkers; D006023:Glycoproteins; C546873:Ibsp protein, rat; D058574:Integrin-Binding Sialoprotein; D012795:Sialoglycoproteins; C512787:bone acidic glycoprotein-75, rat; D000469:Alkaline Phosphatase