Identification and characterization of cellular binding proteins (receptors) for recombinant human bone morphogenetic protein 2B, an initiator of bone differentiation cascade

Vishwas M. Paralkar, R. Glenn Hammonds, A Hari Reddi

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

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 26-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 β 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 El 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 El cells. These data demonstrate specific, high-affinity cell-surface binding proteins for BMP 2B.

Original languageEnglish (US)
Pages (from-to)3397-3401
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number8
StatePublished - 1991
Externally publishedYes

Fingerprint

Bone Morphogenetic Protein 4
Carrier Proteins
Bone and Bones
NIH 3T3 Cells
Platelet-Derived Growth Factor
Transforming Growth Factors
Somatomedins
Fibroblast Growth Factor 2
Osteoblasts
Epidermal Growth Factor
Osteogenesis
Heparin
Membrane Proteins
Fibroblasts
Binding Sites
Cell Line
Temperature

Keywords

  • Bone cells
  • Cartilage
  • Development
  • Morphogenesis
  • Transforming growth factor β superfamily

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

@article{53acf7f6051a42dab42c91776269a2b1,
title = "Identification and characterization of cellular binding proteins (receptors) for recombinant human bone morphogenetic protein 2B, an initiator of bone differentiation cascade",
abstract = "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 26-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 β 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 El 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 El cells. These data demonstrate specific, high-affinity cell-surface binding proteins for BMP 2B.",
keywords = "Bone cells, Cartilage, Development, Morphogenesis, Transforming growth factor β superfamily",
author = "Paralkar, {Vishwas M.} and Hammonds, {R. Glenn} and Reddi, {A Hari}",
year = "1991",
language = "English (US)",
volume = "88",
pages = "3397--3401",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "8",

}

TY - JOUR

T1 - Identification and characterization of cellular binding proteins (receptors) for recombinant human bone morphogenetic protein 2B, an initiator of bone differentiation cascade

AU - Paralkar, Vishwas M.

AU - Hammonds, R. Glenn

AU - Reddi, A Hari

PY - 1991

Y1 - 1991

N2 - 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 26-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 β 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 El 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 El cells. These data demonstrate specific, high-affinity cell-surface binding proteins for BMP 2B.

AB - 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 26-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 β 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 El 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 El cells. These data demonstrate specific, high-affinity cell-surface binding proteins for BMP 2B.

KW - Bone cells

KW - Cartilage

KW - Development

KW - Morphogenesis

KW - Transforming growth factor β superfamily

UR - http://www.scopus.com/inward/record.url?scp=0026351807&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026351807&partnerID=8YFLogxK

M3 - Article

C2 - 1849655

AN - SCOPUS:0026351807

VL - 88

SP - 3397

EP - 3401

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 8

ER -