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Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate

Heparan sulfate (HS) is a polysaccharide known to modulate many important biological processes, including Wnt signaling. However, the biochemical interaction between HS and Wnt molecules is not well characterized largely due to the lack of suitable methods. To determine the Wnt binding domain in HS,...

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Autores principales: Gao, Wei, Xu, Yongmei, Liu, Jian, Ho, Mitchell
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4869111/
https://www.ncbi.nlm.nih.gov/pubmed/27185050
http://dx.doi.org/10.1038/srep26245
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author Gao, Wei
Xu, Yongmei
Liu, Jian
Ho, Mitchell
author_facet Gao, Wei
Xu, Yongmei
Liu, Jian
Ho, Mitchell
author_sort Gao, Wei
collection PubMed
description Heparan sulfate (HS) is a polysaccharide known to modulate many important biological processes, including Wnt signaling. However, the biochemical interaction between HS and Wnt molecules is not well characterized largely due to the lack of suitable methods. To determine the Wnt binding domain in HS, we used a Wnt signaling-inhibitory antibody (HS20) and a panel of synthetic HS oligosaccharides with distinct lengths and sulfation modifications. We found that the binding of HS20 to heparan sulfate required sulfation at both the C2 position (2-O-sulfation) and C6 position (6-O-sulfation). The oligosaccharides with the greatest competitive effect for HS20 binding were between six and eight saccharide residues in length. Additionally, a four residue-long oligosaccharide could also be recognized by HS20 if an additional 3-O-sulfation modification was present. Furthermore, similar oligosaccharides with 2-O, 6-O and 3-O-sulfations showed inhibition for Wnt activation. These results have revealed that HS20 and Wnt recognize a HS structure containing IdoA2S and GlcNS6S, and that the 3-O-sulfation in GlcNS6S3S significantly enhances the binding of both HS20 and Wnt. This study provides the evidence for identifying the Wnt binding domain in HS and suggests a therapeutic approach to target the interaction of Wnt and HS in cancer and other diseases.
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spelling pubmed-48691112016-06-01 Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate Gao, Wei Xu, Yongmei Liu, Jian Ho, Mitchell Sci Rep Article Heparan sulfate (HS) is a polysaccharide known to modulate many important biological processes, including Wnt signaling. However, the biochemical interaction between HS and Wnt molecules is not well characterized largely due to the lack of suitable methods. To determine the Wnt binding domain in HS, we used a Wnt signaling-inhibitory antibody (HS20) and a panel of synthetic HS oligosaccharides with distinct lengths and sulfation modifications. We found that the binding of HS20 to heparan sulfate required sulfation at both the C2 position (2-O-sulfation) and C6 position (6-O-sulfation). The oligosaccharides with the greatest competitive effect for HS20 binding were between six and eight saccharide residues in length. Additionally, a four residue-long oligosaccharide could also be recognized by HS20 if an additional 3-O-sulfation modification was present. Furthermore, similar oligosaccharides with 2-O, 6-O and 3-O-sulfations showed inhibition for Wnt activation. These results have revealed that HS20 and Wnt recognize a HS structure containing IdoA2S and GlcNS6S, and that the 3-O-sulfation in GlcNS6S3S significantly enhances the binding of both HS20 and Wnt. This study provides the evidence for identifying the Wnt binding domain in HS and suggests a therapeutic approach to target the interaction of Wnt and HS in cancer and other diseases. Nature Publishing Group 2016-05-17 /pmc/articles/PMC4869111/ /pubmed/27185050 http://dx.doi.org/10.1038/srep26245 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Gao, Wei
Xu, Yongmei
Liu, Jian
Ho, Mitchell
Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
title Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
title_full Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
title_fullStr Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
title_full_unstemmed Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
title_short Epitope mapping by a Wnt-blocking antibody: evidence of the Wnt binding domain in heparan sulfate
title_sort epitope mapping by a wnt-blocking antibody: evidence of the wnt binding domain in heparan sulfate
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4869111/
https://www.ncbi.nlm.nih.gov/pubmed/27185050
http://dx.doi.org/10.1038/srep26245
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