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Construction of heparan sulfate microarray for investigating the binding of specific saccharide sequences to proteins.

Authors
  • Horton, Maurice1
  • Su, Guowei1
  • Yi, Lin1
  • Wang, Zhangjie1
  • Xu, Yongmei1
  • Pagadala, Vijayakanth2
  • Zhang, Fuming3
  • Zaharoff, David A4
  • Pearce, Ken1
  • Linhardt, Robert J3
  • Liu, Jian1
  • 1 Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina, Chapel Hill, NC, USA.
  • 2 Glycan Therapeutics, LLC, 617 Hutton Street, Raleigh, NC, USA.
  • 3 Department of Chemistry and Chemical Biology, Center for Biotechnology and interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.
  • 4 Joint Department of Biomedical Engineering, University of North Carolina-Chapel Hill & North Carolina State University, Raleigh, NC, USA.
Type
Published Article
Journal
Glycobiology
Publisher
Oxford University Press
Publication Date
Apr 01, 2021
Volume
31
Issue
3
Pages
188–199
Identifiers
DOI: 10.1093/glycob/cwaa068
PMID: 32681173
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Heparan sulfate (HS) is a heterogeneous, extracellular glycan that interacts with proteins and other molecules affecting many biological processes. The specific binding motifs of HS interactions are of interest, but have not been extensively characterized. Glycan microarrays are valuable tools that can be used to probe the interactions between glycans and their ligands while relying on relatively small amounts of samples. Recently, chemoenzymatic synthesis of HS has been employed to produce specific HS structures that can otherwise be difficult to produce. In this study, a microarray of diverse chemoenzymatically synthesized HS structures was developed and HS interactions were characterized. Fluorescently labeled antithrombin III (AT) and fibroblast growth factor-2 (FGF2) were screened against 95 different HS structures under three different printing concentrations to confirm the utility of this microarray. Specific sulfation patterns were found to be important for binding to these proteins and results are consistent with previous specificity studies. Furthermore, the binding affinities (KD,surf) of AT and FGF2 to multiple HS structures were determined using a microarray technique and is consistent with previous reports. Lastly, the 95-compound HS microarray was used to determine the distinct binding profiles for interleukin 12 and platelet factor 4. This technique is ideal for rapid expansion and will be pivotal to the high-throughput characterization of biologically important structure/function relationships. © The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: [email protected]

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