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Efficient platform for synthesizing comprehensive heparan sulfate oligosaccharide libraries for decoding glycosaminoglycan–protein interactions

Nature Chemistry volume 15, pages 1108–1117 (2023)Cite this article

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Abstract

Glycosaminoglycans (GAGs) are abundant, ubiquitous carbohydrates in biology, yet their structural complexity has limited an understanding of their biological roles and structure–function relationships. Synthetic access to large collections of well defined, structurally diverse GAG oligosaccharides would provide critical insights into this important class of biomolecules and represent a major advance in glycoscience. Here we report a new platform for synthesizing large heparan sulfate (HS) oligosaccharide libraries displaying comprehensive arrays of sulfation patterns. Library synthesis is made possible by improving the overall synthetic efficiency through universal building blocks derived from natural heparin and a traceless fluorous tagging method for rapid purification with minimal manual manipulation. Using this approach, we generated a complete library of 64 HS oligosaccharides displaying all possible 2-O-, 6-O- and N-sulfation sequences in the tetrasaccharide GlcN–IdoA–GlcN–IdoA. These diverse structures provide an unprecedented view into the sulfation code of GAGs and identify sequences for modulating the activities of important growth factors and chemokines.

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Fig. 1: Synthesis of a comprehensive HS tetrasaccharide library.
Fig. 2: Synthesis of universal building block 65.
Fig. 3: Divergent synthesis of the HS tetrasaccharide library.
Fig. 4: FGF2 recognizes specific HS sulfation sequences.
Fig. 5: Comparison of binding of selected glycans to FGF2.
Fig. 6: HS sulfation specificities of FGF4 and CXCL8.

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All data associated with this report are contained within the manuscript or the Supplementary Information. Source data are provided with this Paper.

Code availability

All code associated with this report is publicly available at https://doi.org/10.5281/zenodo.7787616.

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Acknowledgements

Financial support was provided by the National Institutes of Health (NIH U01 GM116262 to L.C.H.-W. and X.H., R44 GM134738 to V. Pagadala, L.C.H.-W. and X.H., U01 GM116248 to N.L.B.P., and T32 GM109825 and T32 GM131994 to M.K.K.) and the National Science Foundation (DGE-1745301 to A.W.S.). We thank M. Shahgholi in the CCE Division Mass Spectrometry Facility and D. Vander Velde in the CCE Division NMR Facility at Caltech for technical support.

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Authors and Affiliations

  1. Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA, USA

    Lei Wang, Alexander W. Sorum, Bo-Shun Huang, Nitin Pawar & Linda C. Hsieh-Wilson

  2. Department of Chemistry, Indiana University, Bloomington, IN, USA

    Mallory K. Kern & Nicola L. B. Pohl

  3. Glycan Therapeutics Corp, Raleigh, NC, USA

    Guowei Su

  4. Departments of Chemistry and Biomedical Engineering, Institute for Quantitative Health Science and Engineering, Michigan State University, East Lansing, MI, USA

    Xuefei Huang

  5. Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA

    Jian Liu

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Contributions

L.W., A.W.S., B.-S.H., N.P., X.H., J.L., N.L.B.P. and L.C.H.-W. designed the research. L.W., A.W.S., B.-S.H., M.K.K. and G.S. performed the research. L.W., A.W.S., B.-S.H. and L.C.H.-W. wrote the paper.

Corresponding author

Correspondence to Linda C. Hsieh-Wilson.

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Competing interests

J.L. is a founder and chief scientific officer of Glycan Therapeutics. G.S. is an employee of Glycan Therapeutics and has stock options from the company. The remaining authors declare no competing interests.

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Nature Chemistry thanks Hongzhi Cao, Chi-Huey Wong and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary Information

Supplementary Figs. 1–8, Tables 1 and 2, synthetic procedures and characterization, and NMR spectra.

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Source data

Source Data Fig. 4

Microarray data provided as fluorescence signal for FGF2.

Source Data Fig. 6

Microarray data provided as fluorescence signal for FGF4.

Source Data Fig. 6

Microarray data provided as fluorescence signal for CXCL8.

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Wang, L., Sorum, A.W., Huang, BS. et al. Efficient platform for synthesizing comprehensive heparan sulfate oligosaccharide libraries for decoding glycosaminoglycan–protein interactions. Nat. Chem. 15, 1108–1117 (2023). https://doi.org/10.1038/s41557-023-01248-4

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