[Purification of bone morphogenetic protein and investigation of its effects on osteoblastic cell line UMR108].
It is well known that the bone matrix contains proteins which can induce ectopic endochondral bone formation in vivo. One class of these proteins is the bone morphogenetic protein (BMP). In order to investigate the physiological function of the BMP, its purification was attempted from an extract of demineralized bone matrix and its actions on the osteoblastic cell line were investigated. To isolate the BMP, a demineralized bone matrix was extracted with 4M guanidine-HCl. A water-insoluble fraction (G-WI) was separated from the demineralized bone extract by dialysis against distilled water and centrifugation. The BMP was purified from G-WI by gel filtration on Sephacryl S-200 HR, cation exchange with Mono-S, heparin affinity column and finally by C1/8 reverse phase chromatography. Peptide sequence analysis revealed that the purified BMP fraction contained "BMP-3" reported by Wozney et al. (1988). In order to investigate its function, the BMP was applied to the rat osteogenic sarcoma cell line UMR108. The BMP inhibited the growth of the UMR108 cells and enhanced the alkaline phosphatase activity in a dose-responsive manner.
Sakurai
N
N
Department of Oral and Maxillofacial Surgery II, Faculty of Dentistry, Tokyo Medical and Dental University.
D000595:Amino Acid Sequence; D000818:Animals; D001857:Bone Matrix / Q000737:chemistry; D019485:Bone Morphogenetic Proteins; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006133:Growth Substances / Q000302:isolation & purification* / Q000494:pharmacology*; D006801:Humans; D008969:Molecular Sequence Data; D010006:Osteoblasts / Q000187:drug effects*; D012516:Osteosarcoma / Q000473:pathology; D011506:Proteins / Q000302:isolation & purification* / Q000494:pharmacology*; D051381:Rats; D014407:Tumor Cells, Cultured
D019485:Bone Morphogenetic Proteins; D006133:Growth Substances; D011506:Proteins