Identification and characterization of cellular binding proteins (receptors) for recombinant human bone morphogenetic protein 2B, an initiator of bone differentiation cascade.
Bone morphogenetic protein 2B (BMP 2B), is a heparin-binding bone differentiation factor that initiates endochondral bone formation in rats when implanted subcutaneously. The molecular mechanism of action of this differentiation factor is not known, and as a first step we have examined BMP 2B-responsive cells for the presence of specific cellular binding proteins. Using 125I-labeled BMP 2B, specific high-affinity binding sites for recombinant human BMP 2B on MC3T3 E1 osteoblast-like cells as well as on NIH 3T3 fibroblasts were identified. Platelet-derived growth factor, insulin-like growth factor 1, basic fibroblast growth factor, epidermal growth factor, and transforming growth factor beta did not compete for the binding of radiolabeled BMP 2B. The binding of BMP 2B is a time- and temperature-dependent process. Chemical crosslinking of radiolabeled BMP showed two components (apparent size, 200 and 70 kDa in MC3T3 E1 cells and 200 and 90 kDa in NIH 3T3 cells). A minor component at 60 kDa was also detected in both cell lines. Scatchard analysis of the binding data showed a high-affinity receptor with an apparent dissociation constant of 128 +/- 40 pM in MC3T3 E1 cells. These data demonstrate specific, high-affinity cell-surface binding proteins for BMP 2B.
Paralkar
V M
VM
Bone Cell Biology Section, National Institute of Dental Research, National Institutes of Health, Bethesda, MD 20892.
Hammonds
R G
RG
Reddi
A H
AH
D000345:Affinity Labels; D000818:Animals; D019485:Bone Morphogenetic Proteins; D001842:Bone and Bones / Q000166:cytology / Q000378:metabolism*; D002460:Cell Line; D003432:Cross-Linking Reagents; D004247:DNA / Q000096:biosynthesis; D006801:Humans; D051379:Mice; D009024:Morphogenesis; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013696:Temperature; D013997:Time Factors
D000345:Affinity Labels; D019485:Bone Morphogenetic Proteins; D003432:Cross-Linking Reagents; D011506:Proteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; D004247:DNA