Skip to main content
SpringerLink
Log in

A novel human bone morphogenetic protein-7 variant with an enriched heparin-binding site

  • Cell Molecular Biology
  • Published:
Molecular Biology Aims and scope Submit manuscript

Abstract

BMPs are osteoinductive proteins which are used in treatment of acute fractures. Large quantities of recombinant proteins are usually needed to achieve efficacy in the clinic. This translates to severe complications and high costs. Different strategies have been developed to improve the efficacy and safety of BMPs. Modification of the heparin-binding site in order to increase the local retention time of the morphogen is one of these approaches. Aiming at further improvement in properties of BMP-7, a novel form of this protein was designed and expressed successfully in Chinese Hamster Ovarian (CHO) cells. Substitution of the Bone morphogenetic protein-7 N-terminus by the heparin-binding site of Bone morphogenetic protein-2 was carried out to increase the heparin binding capacity of the novel protein. It was found that the novel variant, retained its in vitro biological activity and the heparin binding capacity of this protein was approximately 20% higher than that of the wild-type at a protein concentration of 100 ng/mL. The novel protein as the first variant of hBMP-7 with the enriched heparin-binding site may offer more advantages in clinical use as compared to the existing commercial form.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Kirker-Head C.A. 2000. Potential applications and delivery strategies for bone morphogenetic proteins. Adv. Drug Deliv. Rev. 43, 65–92.

    Article  PubMed  CAS  Google Scholar 

  2. Swencki-Underwood B., Mills J.K., Vennarini J., et al. 2008. Expression and characterization of a human BMP-7 variant with improved biochemical properties. Protein Expr. Purif. 57, 312–319.

    Article  PubMed  CAS  Google Scholar 

  3. Dinopoulos H., Giannoudis P.V. 2007. (iv) The use of bone morphogenetic proteins (BMPs) in long-bone non-unions. Curr. Orthopaed. 21, 268–279.

    Article  Google Scholar 

  4. Anitua E., Sánchez M., Orive G., Andia I. 2008. Delivering growth factors for therapeutics. Trends Pharmacol. Sci. 29, 37–41.

    Article  PubMed  CAS  Google Scholar 

  5. Jeon O., Song S.J., Yang H.S., Bhang S.-H., Kang S.-W., Sung M.A., Lee J H., Kim B.-S. 2008. Long-term delivery enhances in vivo osteogenic efficacy of bone morphogenetic protein-2 compared to short-term delivery. Biochem. Biophys. Res. Commun. 369, 774–780.

    Article  PubMed  CAS  Google Scholar 

  6. Zhu W., Kim J., Cheng C., Rawlins B.A., Boachie-Adjei O., Crystal R.G., Hidaka C. 2006. Noggin regulation of bone morphogenetic protein (BMP) 2/7 heterodimer activity in vitro. Bone. 39, 61–71.

    Article  PubMed  CAS  Google Scholar 

  7. Han D., Liu W., Ao Q., Wang G. 2008. Optimal delivery systems for bone morphogenetic proteins in orthopedic applications should model initial tissue repair structures by using a heparin-incorporated fibrin-fibronectin matrix. Med. Hypotheses. 71, 374–378.

    Article  PubMed  CAS  Google Scholar 

  8. Kübler N., Würzler K., Reuther J., Sieber E., Kirchner T., Seebald W. 2000. The influence of various factors on the bone forming properties of recombinant BMPs. Mund Kiefer Gesichtschir. 4, 465–469.

    Article  Google Scholar 

  9. Takada T., Katagiri T., Ifuku M., Morimura N., Kobayashi M., Hasegawa K., Ogamo A., Kamijo R. 2003. Sulfated polysaccharides enhance the biological activities of bone morphogenetic proteins. J. Biol. Chem. 278, 43229.

    Article  PubMed  CAS  Google Scholar 

  10. Irie A., Takami M., Kubo H., Sekino-Suzuki N., Kasahara K., Sanai Y. 2007. Heparin enhances osteoclastic bone resorption by inhibiting osteoprotegerin activity. Bone. 41, 165–174.

    Article  PubMed  CAS  Google Scholar 

  11. Lin H., Zhao Y., Sun W., Chen B., Zhang J., Zhao W., Xiao Z., Dai J. 2008. The effect of crosslinking heparin to demineralized bone matrix on mechanical strength and specific binding to human bone morphogenetic protein-2. Biomaterials. 29, 1189–1197.

    Article  PubMed  CAS  Google Scholar 

  12. Schmoekel H.G., Weber F.E., Schense J.C., Grätz K.W., Schawalder P., Hubbell J.A. 2005. Bone repair with a form of BMP-2 engineered for incorporation into fibrin cell ingrowth matrices. Biotechnol. Bioeng. 89, 253–262.

    Article  PubMed  CAS  Google Scholar 

  13. Johnson Z., Proudfoot A., Handel T. 2005. Interaction of chemokines and glycosaminoglycans: a new twist in the regulation of chemokine function with opportunities for therapeutic intervention. Cytokine Growth Factor Rev. 16, 625–636.

    Article  PubMed  CAS  Google Scholar 

  14. Najjam S., Mulloy B., Theze J., Gordon M., Gibbs R., Rider C.C. 1998. Further characterization of the binding of human recombinant interleukin 2 to heparin and identification of putative binding sites. Glycobiology. 8, 509.

    Article  PubMed  CAS  Google Scholar 

  15. Ruppert R., Hoffmann E., Sebald W. 1996. Human bone morphogenetic protein 2 contains a heparinbinding site which modifies its biological activity. Eur. J. Biochem. 237, 295–302.

    Article  PubMed  CAS  Google Scholar 

  16. Würzler K., Emmert J., Eichelsbacher F., Kübler N., Sebald W., Reuther J. 2004. Evaluation of osteoinductive potency of genetically modified BMP-2 variations. J. Oral. Maxillofac. Surg. 8, 83–92.

    Google Scholar 

  17. Depprich R., Handschel J., Sebald W., Kübler N., Würzler K. 2005. Comparison of the osteogenic activity of bone morphogenetic protein (BMP) mutants. J. Oral. Maxillofac. Surg. 9, 363–368.

    CAS  Google Scholar 

  18. Kübler N., Würzler K., Reuther J., Faller G., Sieber E., Kirchner T., Sebald W. 1999. EHBMP-2: the first BMP-variant with osteoinductive properties. Mund Kiefer Gesichtschir. 3, 134–139.

    Article  Google Scholar 

  19. Vukicevic S., Sampath K.T. 2002. Bone Morphogenetic Proteins: From Laboratory to Clinical Practice (Progress in Inflammation Research). Basel, Switzerland: Birkhäuser.

    Book  Google Scholar 

  20. Irie A., Habuchi H., Kimata K., Sanai Y. 2003. Heparan sulfate is required for bone morphogenetic protein-7 signaling. Biochem. Biophys. Res. Commun. 308, 858–865.

    Article  PubMed  CAS  Google Scholar 

  21. Guermeur Y. 1997. Combinaison de classifieurs statistiques: application a la prediction de structure secondaire des proteines., Ph.D. Thesis, University of Paris 6.

    Google Scholar 

  22. Mahoney D.J., Whittle J.D., Milner C.M., Clark S.J., Mulloy B., Buttle D.J., Jones G.C., Day A.J., Short R.D. 2004. A method for the non-covalent immobilization of heparin to surfaces. Anal. Biochem. 330, 123–129.

    Article  PubMed  CAS  Google Scholar 

  23. Di Liddo R., Grandi C., Venturini M., Dalzoppo D., Negro A., Conconi M., Parnigotto P. 2010. Recombinant human TAT-OP1 to enhance NGF neurogenic potential: preliminary studies on PC12 cells. Protein Eng. Des. Sel. 23, 889.

    Article  PubMed  Google Scholar 

  24. Israel D.I., Nove J., Kerns K.M., Moutsatsos I.K., Kaufman R.J. 1992. Expression and characterization of bone morphogenetic protein-2 in Chinese hamster ovary cells. Growth Factors. 7, 139–150.

    Article  PubMed  CAS  Google Scholar 

  25. Sampath T.K., Maliakal J., Hauschka P., et al. 1992. Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro. J. Biol. Chem. 267, 20352.

    PubMed  CAS  Google Scholar 

  26. Bustos-Valenzuela J., Halcsik E., Bassi E., Demasi M., Granjeiro J., Sogayar M. 2010. Expression, purification, bioactivity, and partial characterization of a recombinant human bone morphogenetic protein-7 produced in human 293T cells. Mol. Biotechnol. 46, 118–126.

    Article  PubMed  CAS  Google Scholar 

  27. Zhao B., Katagiri T., Toyoda H., et al. 2006. Heparin potentiates the in vivo ectopic bone formation induced by bone morphogenetic protein-2. J. Biol. Chem. 281, 23246–23253.

    Article  PubMed  CAS  Google Scholar 

  28. Najjam S., Gibbs R.V., Gordon M.Y., Rider C.C. 1997. Characterization of human recombinant interleukin 2 binding to heparin and heparan sulfate using an ELISA approach. Cytokine. 9, 1013–1022.

    Article  PubMed  CAS  Google Scholar 

  29. Fisher M.C., Li Y., Seghatoleslami M.R., Dealy C.N., Kosher R.A. 2006. Heparan sulfate proteoglycans including syndecan-3 modulate BMP activity during limb cartilage differentiation. Matrix Biol. 25, 27–39.

    Article  PubMed  CAS  Google Scholar 

  30. Reddi A.H. 2001. Interplay between bone morphogenetic proteins and cognate binding proteins in bone and cartilage development: noggin, chordin and DAN. Arthritis Res. 3, 1–5.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Ave., Pasteur square, Southern Kargar Ave., Tehran, Iran

    L. Nematollahi, F. Mahboudi, A. Rahimpour, H. Jahandar & V. Khalaj

Authors
  1. L. Nematollahi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. F. Mahboudi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. A. Rahimpour
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. H. Jahandar
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. V. Khalaj
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to V. Khalaj.

Additional information

Published in Russian in Molekulyarnaya Biologiya, 2013, Vol. 47, No. 3, pp. 453–460.

The article is published in the original.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nematollahi, L., Mahboudi, F., Rahimpour, A. et al. A novel human bone morphogenetic protein-7 variant with an enriched heparin-binding site. Mol Biol 47, 399–405 (2013). https://doi.org/10.1134/S0026893313030096

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1134/S0026893313030096

Keywords

Search

Navigation