Construction of heparan sulfate microarray for investigating the binding of specific saccharide sequences to proteins
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Authors
Horton, Maurice
Su, Guowei
Yi, Lin
Wang, Zhangjie
Xu, Yongmei
Pagadala, Vijayakanth
Zhang, Fuming
Zaharoff, David A.
Pearce, Ken
Linhardt, Robert J.
Issue Date
2021-03-01
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
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.
Description
Glycobiology, 31, 188–199
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
Construction of heparan sulfate microarray for investigating the binding of specific saccharide sequences to proteins, M. Horton, G. Su, L. Yi, Y. Xu, V. Pagadala, F. Zhang, K. Pierce, R. J Linhardt, J. Liu, Glycobiology, 31, 188–199, 2021.
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Journal
Volume
Issue
PubMed ID
DOI
ISSN
14602423
9596658
9596658