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Purification of Bovine Bone Morphogenetic Protein by Hydroxyapatite Chromatography

Urist, M. R. and Huo, Y. K. and Brownell, A. G. and Hohl, W. M. and Buyske, J. and Lietze, A. and Tempst, P. and Hunkapiller, M. and DeLange, R. J. (1984) Purification of Bovine Bone Morphogenetic Protein by Hydroxyapatite Chromatography. Proceedings of the National Academy of Sciences of the United States of America, 81 (2). pp. 371-375. ISSN 0027-8424. PMCID PMC344678. doi:10.1073/pnas.81.2.371.

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Bovine bone morphogenetic protein (bBMP) induces differentiation of mesenchymal-type cells into cartilage and bone. bBMP has an apparent Mr of 18,500 ± 500 and represents <0.001% of the wet weight of bone tissue. A Mr 34,000 protein resembling osteonectin is separated by extraction with Triton X-100. A Mr 24,000 protein and about half of a Mr 22,000 protein are disassociated from bBMP by precipitation in 1.5 M guanidine hydrochloride. Aggregates of bBMP and a Mr 14,000 protein are insoluble in aqueous media; the bBMP becomes soluble when the Mr 14,000 protein is disassociated in 6 M urea and removed from the solution by ultrafiltration. Three separate molecular species with apparent Mrs 18,500, 17,500, and 17,000 are eluted at 0.10, 0.15, and 0.20 M phosphate ion concentrations, respectively, from a hydroxy-apatite column. The Mr 18,500 protein has the amino acid composition of acidic polypeptide and includes four halfcystine residues; the pI is 4.9-5.1. The Mr 22,000 component is a chromoprotein resembling ferritin. The NH2-terminal amino acid sequence of the Mr 17,500 protein simulates histone H2B. The Mr 17,000 protein may possess calmodulin activity. Aggregates of the Mr 18,500 and other proteins induce formation of large deposits of bone; the Mr 18,500 protein alone is rapidly absorbed and induces formation of small deposits. None of the other proteins induces bone formation.

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Additional Information:© 1984 by the National Academy of Sciences Communicated by C. H. Sawyer, September 26, 1983 The authors are indebted to Lynette Knight, B. Wakim, Edward King, K. Sato, A. B. Pretsky, S. J. Krishel, M. Rivera, and L. King for excellent technical assistance. This research was aided by grants from the U.S. Public Health Service and the National Institutes of Health, NIDR DE02103, William M. Keck Foundation, Solo Cup Foundation, and the Stryker Corporation. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funding AgencyGrant Number
W. M. Keck FoundationUNSPECIFIED
Solo Cup FoundationUNSPECIFIED
Stryker CorporatonUNSPECIFIED
Subject Keywords:cell differentiation; osteogenesis; development
Issue or Number:2
PubMed Central ID:PMC344678
Record Number:CaltechAUTHORS:URIpnas84
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:5753
Deposited By: Archive Administrator
Deposited On:31 Oct 2006
Last Modified:08 Nov 2021 20:28

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