Spontaneously Restoring Specific Bioaffinity of RGD in Linear RGD-containing Peptides by Conjugation with Zwitterionic Dendrimers

Liangbo Xu, Nan Xu, Longgang Wang, Haofeng Qian, Yihan Li, Mandi Fang, Ziyin Xiang, Weifeng Lin, Fanxing Zhang, Qing Shao, Matthew T. Bernards, Yao Shi, Yi He, Shengfu Chen

Research output: Contribution to journalArticlepeer-review

Abstract

Peptides are more versatile than small molecule drugs, but their specific bioaffinities are usually lower than their original native proteins because of the loss of preferred conformations. To overcome this key obstacle, we demonstrated a hydrogen bond-induced conformational constraint method to enhance the specific bioaffinities of peptides to achieve a high success rate by using linear RGD-containing peptides as a model of bioactive peptides. By performing molecular simulation, we found that the chemically immobilized linear CRGDS via cysteine (C) at the N-terminus on zwitterionic PAMAM G-5 can not only spontaneously restore the natural conformation of the RGD segment through the assistance of the dynamic hydrogen bond from serine (S) at the C-terminus of the peptide, but it can also narrow the distribution of all possible conformations. Consequently, the conjugates showed comparable or even better high affinity than native proteins without the use of conventional, labor-intensive, synthesis-based structure search methods to construct a binding conformation. In addition, the conjugates showed globular protein-like characteristics chemically, physically, and physiologically. They exhibited not only high efficacy and biosafety both in vitro and in vivo, but they also showed extremely high thermostability even upon boiling in a solution. This approach offers great design flexibility for reviving functional peptides without impairing their high specific affinity for their targets. Statement of significance: In this work, we developed a swift approach to spontaneously restore the natural conformation of a linear peptide from a nature protein and thus enhance its specific bioaffinity instead of constructing a binding conformation by the labor-intensive, synthesis-based structure search method. In details, our new approach involves dynamically constraining the linear peptide on a zwitterionic PAMAM G-5 surface by a combination of chemical bonding at one terminus and dynamic hydrogen bonding at the other terminus of the linear peptide. The zwitterionic background offers abundant interaction sites for hydrogen bonding as well as resistance to nonspecific interactions. This approach fully restores the specific bioaffinity of RGD segments on a zwitterionic PAMAM G-5 through only one conjugation point at the C-terminus of the peptide. Moreover, the bioaffinity of all three types of RGD-containing peptides is successfully restored, which indicates the high rate of success of this approach in affinity restoring.

Original languageEnglish
Pages (from-to)61-72
Number of pages12
JournalActa Biomaterialia
Volume148
DOIs
StatePublished - Aug 2022

Bibliographical note

Funding Information:
We thank Dr. Shaoyi Jiang (Cornell University) and Dr. François Baneyx (University of Washington, Seattle) for helpful discussion and comments on the manuscript. This work is supported by the National Natural Science Foundation of China (grant number 21674092, 21474085, 51933009,21676245, and 51750110495), and the National Key Research and Development Program of China (grant number 2017YFB0702502).

Funding Information:
The data supporting this work are available in the article and its Supplementary Information files. Additional raw data is available from the authors upon reasonable request. Supplementary discussion, supplementary Figs. 1 to 9, and supplementary Tables 1 and 2 are available in the supplementary material of this article. We thank Dr. Shaoyi Jiang (Cornell University) and Dr. François Baneyx (University of Washington, Seattle) for helpful discussion and comments on the manuscript. This work is supported by the National Natural Science Foundation of China (grant number 21674092, 21474085, 51933009,21676245, and 51750110495), and the National Key Research and Development Program of China (grant number 2017YFB0702502).

Publisher Copyright:
© 2022

Keywords

  • Conformational constraint
  • High efficacy and biosafety
  • High thermostability
  • Molecular simulation
  • RGD/dendrimers conjugate

ASJC Scopus subject areas

  • Biotechnology
  • Biomaterials
  • Biochemistry
  • Biomedical Engineering
  • Molecular Biology

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