Abstract
Biological systems have evolved to utilize numerous proteins with capacity to bind polysaccharides for the purpose of optimizing their function. A well-known subset of these proteins with binding domains for the highly diverse sulfated polysaccharides are important growth factors involved in biological development and tissue repair. We report here on supramolecular sulfated glycopeptide nanostructures, which display a trisulfated monosaccharide on their surfaces and bind five critical proteins with different polysaccharide-binding domains. Binding does not disrupt the filamentous shape of the nanostructures or their internal β-sheet backbone, but must involve accessible adaptive configurations to interact with such different proteins. The glycopeptide nanostructures amplified signalling of bone morphogenetic protein 2 significantly more than the natural sulfated polysaccharide heparin, and promoted regeneration of bone in the spine with a protein dose that is 100-fold lower than that required in the animal model. These highly bioactive nanostructures may enable many therapies in the future involving proteins.
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Acknowledgements
Funding was provided by the NIH National Institute of Dental and Craniofacial Research grant 5R01DE015920-10, and also by the Louis A. Simpson & Kimberly Querrey Center for Regenerative Nanomedicine at Northwestern University. The synthesis and structural characterization of this work was supported by the Center for Bio-Inspired Energy Science, an Energy Frontier Research Center funded by DOE, Office of Science, Basic Energy Sciences, under award no. DE-SC0000989. Studies on the dynamics and X-ray scattering were supported by the US Department of Energy (DOE), Office of Science, Office of Basic Energy Sciences, under award no. DE-FG02-00ER45810. The SAXS experiments were performed at the DND-CAT located at Sector 5 of the Advanced Photon Source (APS). Synchrotron X-ray μCT experiments were performed at Sector 2-BM of the APS. We thank the following facilities at Northwestern University: Analytical BioNanotechnology Equipment Core, Peptide Synthesis Core, Center for Advanced Microscopy, Biological Imaging Facility, Keck Biophysics Facility, Integrated Molecular Structure Education and Research Center, Quantitative Bio-element Imaging Center, Center for Advanced Molecular Imaging, and Research Histology and Phenotyping Laboratory. The Biophysics Core Facility at the University of Chicago was also used. Refer to the Supplementary Information for facilities support. S.S.L. thanks the Samsung Scholarship. We are grateful to S. Weigand for assistance with the X-ray scattering, D. Sebald (University of Würzburg, Germany) and A. Lander (University of California-Irvine, USA) for a generous gift of EHBMP-2, to D. Ornitz (Washington University, Saint Louis, USA) for providing the engineered BaF3 cell line, to L. Palmer and K. Sato for helpful discussions, to C. Haney for assisting with μCT scanning and analyses, and to M. Seniw for molecular graphics. Although not used in this Article, we also thank S. Pshenychnyi and Y. Goo in assisting us with recombinant protein production and proteomics.
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S.S.L., T.F. and S.I.S. conceived the project. S.S.L., T.F., E.S. and Z.A. designed and performed the experiments and analysed the data. F.C., Z.A., D.S.C., J.A.W., R.W.C., R.D.F., M.S.S., K.M.K., A.D.S., J.T.S., C.Y., G.S., S.Z.H., M.T.M. and S.R.S. performed the experiments. Z.Y. assisted in synthesis. S.R.S., W.K.H. and E.L.H. supervised the in vivo study and analysis. S.S.L., T.F., E.L.H. and S.I.S. wrote the manuscript. All authors accepted the final version of the manuscript.
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Lee, S., Fyrner, T., Chen, F. et al. Sulfated glycopeptide nanostructures for multipotent protein activation. Nature Nanotech 12, 821–829 (2017). https://doi.org/10.1038/nnano.2017.109
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DOI: https://doi.org/10.1038/nnano.2017.109
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