The Protein Journal

, Volume 38, Issue 1, pp 12–22 | Cite as

Grafting, Stripping and Stapling of Helical Peptides from the Dimerization Interface of ONFH-Related Bone Morphogenetic Protein-2

  • Wenqi Song
  • Kunzheng WangEmail author
  • Wei Wang
  • Pei Yang
  • Xiaoqian Dang


Transforming growth factor-β/bone morphogenetic protein (TGF-β/BMP) signaling plays a fundamental role in embryonic skeletal development and postnatal bone homeostasis. The signaling pivot protein BMP-2 belongs to the TGF-β superfamily and has been implicated in the pathogenesis of osteonecrosis of femoral head (ONFH). The biologically functional BMP-2 is a homodimer that has two tightly packed cores at its dimerization interface; each core is defined by the intermolecular interaction between a helical arm from one monomer and a hydrophobic pocket from another monomer. Inhibition and disruption of BMP-2 dimerization have been recognized as an attractive therapeutic strategy against ONFH. Here, we investigate the self-binding behavior of helical arm-derived peptides to the BMP-2 dimerization interface. The native BMP-2 helical arm and its several grafted versions from BMP-4, BMP-6 and BMP-7 are stripped from the intact dimerization interface to generate a number of isolated helical peptides. Computational simulations demonstrate that the stripping does not substantially influence the direct intermolecular interaction between BMP-2 monomer and these helical peptides or desolvation effect upon the interaction. However, the C-terminus of stripped peptides is found to have an intrinsic disorder and large flexibility in the isolated state, which would impair the rebinding of stripped peptides to BMP-2. Next, we rationally design a hydrocarbon bridge across the C-terminal residues 65 and 69 of helical peptides, which can effectively constrain peptide conformational flexibility in the isolated state, thus considerably promoting the binding potency of stripped helical peptides. Circular dichroism (CD) spectroscopy reveals that the peptide helicity increases from 51.8 to 67.9% upon hydrocarbon stapling. Fluorescence polarization assays substantiate that, as designed, the stapling can convert these helical peptides from weak binders to moderate or good binders of BMP-2 protein; their Kd values are improved by up to ~ fourfold.


Bone morphogenetic protein-2 Helical peptide Peptide-mediated protein–protein interaction Hydrocarbon stapling Osteonecrosis of femoral head 



This work was supported by the National Natural Science Foundation of China (No. 81371962).

Compliance with Ethical Standards

Conflict of interest

The author declares that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Wenqi Song
    • 1
    • 2
  • Kunzheng Wang
    • 1
    Email author
  • Wei Wang
    • 1
  • Pei Yang
    • 1
  • Xiaoqian Dang
    • 1
  1. 1.Department of OrthopedicsThe Second Affiliated Hospital of Xi’an Jiao Tong UniversityXi’anChina
  2. 2.Department of OrthopedicsShanghai Jiao Tong University Affiliated Sixth People’s HospitalShanghaiChina

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