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Title: Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity

Abstract

Glycans possess significant chemical diversity; glycan binding proteins (GBPs) recognize specific glycans to translate their structures to functions in various physiological and pathological processes. Therefore, the discovery and characterization of novel GBPs and characterization of glycan–GBP interactions are significant to provide potential targets for therapeutic intervention of many diseases. Here, we report the biochemical, functional, and structural characterization of a 130-amino-acid protein, Y3, from the mushroomCoprinus comatus. Biochemical studies of recombinant Y3 from a yeast expression system demonstrated the protein is a unique GBP. Additionally, we show that Y3 exhibits selective and potent cytotoxicity toward human T-cell leukemia Jurkat cells compared with a panel of cancer cell lines via inducing caspase-dependent apoptosis. Screening of a glycan array demonstrated GalNAcβ1–4(Fucα1–3)GlcNAc (LDNF) as a specific Y3-binding ligand. Here, to provide a structural basis for function, the crystal structure was solved to a resolution of 1.2 Å, revealing a single-domain αβα-sandwich motif. Two monomers were dimerized to form a large 10-stranded, antiparallel β-sheet flanked by α-helices on each side, representing a unique oligomerization mode among GBPs. A large glycan binding pocket extends into the dimeric interface, and docking of LDNF identified key residues for glycan interactions. Disruption of residues predicted to be involvedmore » in LDNF/Y3 interactions resulted in the significant loss of binding to Jurkat T-cells and severely impaired their cytotoxicity. Collectively, these results demonstrate Y3 to be a GBP with selective cytotoxicity toward human T-cell leukemia cells and indicate its potential use in cancer diagnosis and treatment.« less

Authors:
 [1]; ORCiD logo [2];  [1];  [1];  [1];  [1];  [1];  [3];  [1];  [1]; ORCiD logo [1]
  1. Univ. of Florida, Gainesville, FL (United States)
  2. Univ. of Florida, Gainesville, FL (United States); Harvard Medical School, Boston, MA (United States)
  3. Virginia Commonwealth Univ., Richmond, VA (United States)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1419882
Grant/Contract Number:  
R24 GM098791; P41 GM103694
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 114; Journal Issue: 34; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; glycan binding protein; Coprinus comatus; cytotoxicity; LDNF; crystal structure

Citation Formats

Zhang, Peilan, Li, Kunhua, Yang, Guang, Xia, Changqing, Polston, Jane E., Li, Gengnan, Li, Shiwu, Lin, Zhao, Yang, Li-jun, Bruner, Steven D., and Ding, Yousong. Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity. United States: N. p., 2017. Web. doi:10.1073/pnas.1706894114.
Zhang, Peilan, Li, Kunhua, Yang, Guang, Xia, Changqing, Polston, Jane E., Li, Gengnan, Li, Shiwu, Lin, Zhao, Yang, Li-jun, Bruner, Steven D., & Ding, Yousong. Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity. United States. https://doi.org/10.1073/pnas.1706894114
Zhang, Peilan, Li, Kunhua, Yang, Guang, Xia, Changqing, Polston, Jane E., Li, Gengnan, Li, Shiwu, Lin, Zhao, Yang, Li-jun, Bruner, Steven D., and Ding, Yousong. Mon . "Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity". United States. https://doi.org/10.1073/pnas.1706894114. https://www.osti.gov/servlets/purl/1419882.
@article{osti_1419882,
title = {Cytotoxic protein from the mushroom Coprinus comatus possesses a unique mode for glycan binding and specificity},
author = {Zhang, Peilan and Li, Kunhua and Yang, Guang and Xia, Changqing and Polston, Jane E. and Li, Gengnan and Li, Shiwu and Lin, Zhao and Yang, Li-jun and Bruner, Steven D. and Ding, Yousong},
abstractNote = {Glycans possess significant chemical diversity; glycan binding proteins (GBPs) recognize specific glycans to translate their structures to functions in various physiological and pathological processes. Therefore, the discovery and characterization of novel GBPs and characterization of glycan–GBP interactions are significant to provide potential targets for therapeutic intervention of many diseases. Here, we report the biochemical, functional, and structural characterization of a 130-amino-acid protein, Y3, from the mushroomCoprinus comatus. Biochemical studies of recombinant Y3 from a yeast expression system demonstrated the protein is a unique GBP. Additionally, we show that Y3 exhibits selective and potent cytotoxicity toward human T-cell leukemia Jurkat cells compared with a panel of cancer cell lines via inducing caspase-dependent apoptosis. Screening of a glycan array demonstrated GalNAcβ1–4(Fucα1–3)GlcNAc (LDNF) as a specific Y3-binding ligand. Here, to provide a structural basis for function, the crystal structure was solved to a resolution of 1.2 Å, revealing a single-domain αβα-sandwich motif. Two monomers were dimerized to form a large 10-stranded, antiparallel β-sheet flanked by α-helices on each side, representing a unique oligomerization mode among GBPs. A large glycan binding pocket extends into the dimeric interface, and docking of LDNF identified key residues for glycan interactions. Disruption of residues predicted to be involved in LDNF/Y3 interactions resulted in the significant loss of binding to Jurkat T-cells and severely impaired their cytotoxicity. Collectively, these results demonstrate Y3 to be a GBP with selective cytotoxicity toward human T-cell leukemia cells and indicate its potential use in cancer diagnosis and treatment.},
doi = {10.1073/pnas.1706894114},
url = {https://www.osti.gov/biblio/1419882}, journal = {Proceedings of the National Academy of Sciences of the United States of America},
issn = {0027-8424},
number = 34,
volume = 114,
place = {United States},
year = {2017},
month = {8}
}

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