Presented in abstract form at The American Society for Bone and Mineral Research Annual Meeting, Baltimore, MD, U.S.A., September 1995.
Characterization of Native and Recombinant Bone Sialoprotein: Delineation of the Mineral-Binding and Cell Adhesion Domains and Structural Analysis of the RGD Domain†
Article first published online: 1 AUG 1997
Copyright © 1997 ASBMR
Journal of Bone and Mineral Research
Volume 12, Issue 8, pages 1210–1222, August 1997
How to Cite
Stubbs, J. T., Mintz, K. P., Eanes, E. D., Torchia, D. A. and Fisher, L. W. (1997), Characterization of Native and Recombinant Bone Sialoprotein: Delineation of the Mineral-Binding and Cell Adhesion Domains and Structural Analysis of the RGD Domain. J Bone Miner Res, 12: 1210–1222. doi: 10.1359/jbmr.19220.127.116.110
- Issue published online: 4 DEC 2009
- Article first published online: 1 AUG 1997
- Manuscript Accepted: 31 MAR 1997
- Manuscript Revised: 21 MAR 1997
- Manuscript Received: 19 DEC 1996
Bone sialoprotein is a small, sulfated, and phosphorylated integrin-binding glycoprotein apparently found only in tissues that eventually mineralize. Nondenatured bone sialoprotein (BSP) purified from rat osteosarcoma cell line (UMR 106–01 BSP) culture media is shown to have a hydroxyapatite Kd ≈ 2.6 × 10−9 M, perhaps the strongest affinity for this mineral of any of the matrix proteins. Both native BSP and a 47 kD fragment of UMR-BSP (Fragment 1 ∼133A–∼265Y) are more potent inhibitors of seeded hydroxyapatite crystal growth than recombinant human BSP fragments lacking post-translational modifications. The recombinant proteins, however, do show reproducible inhibitory activity, suggesting that at least some of the strong mineral-binding properties are encoded directly within the protein sequence itself. BSP facilitates the adhesion of several cell types through its integrin binding (RGD) tripeptide sequence. Nuclear magnetic resonance (NMR) analysis of a15N-enriched 59 amino acid recombinant domain containing the RGD tripeptide shows that the structure of this isolated domain is highly flexible with or without 5 mM calcium. Previous work has also shown that an endogenous fragment of UMR-BSP (Fragment 1) supports cell adhesion in the absence of the RGD sequence. In this report, non-RGD cell adhesion sites are localized within conserved amino- and carboxy-terminal tyrosine-rich domains of recombinant human BSP. Given the proximity of the latter non-RGD cell adhesion site to the RGD tripeptide, a model of BSP-receptor interactions is presented.