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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. 2017. "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 = {Mon Aug 07 00:00:00 EDT 2017},
month = {Mon Aug 07 00:00:00 EDT 2017}
}

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Works referenced in this record:

Protein Glycosylation in Cancer
journal, January 2015


Crystal structures of human glutaminyl cyclase, an enzyme responsible for protein N-terminal pyroglutamate formation
journal, August 2005


Apoptosis: A Review of Programmed Cell Death
journal, June 2007


Glycans in cancer and inflammation — potential for therapeutics and diagnostics
journal, June 2005


Antioxidant Properties of Cap and Stipe from Coprinus comatus
journal, March 2010


Antibody Responses to the Fucosylated LacdiNAc Glycan Antigen in Schistosoma mansoni-Infected Mice and Expression of the Glycan among Schistosomes
journal, December 2000


Dali server: conservation mapping in 3D
journal, May 2010


Galectin-1: a small protein with major functions
journal, July 2006


A higher-level phylogenetic classification of the Fungi
journal, May 2007


Recent advances in employing molecular modelling to determine the specificity of glycan-binding proteins
journal, October 2014


From lectin structure to functional glycomics: principles of the sugar code
journal, June 2011


A dual function for a bacterial small RNA: SgrS performs base pairing-dependent regulation and encodes a functional polypeptide
journal, November 2007


Convergent and divergent mechanisms of sugar recognition across kingdoms
journal, October 2014


CD19-targeted chimeric antigen receptor T-cell therapy for acute lymphoblastic leukemia
journal, June 2015


Proteins of higher fungi – from forest to application
journal, May 2012


Investigation of SSEA-4 Binding Protein in Breast Cancer Cells
journal, April 2013


Lectins: Carbohydrate-specific Reagents and Biological Recognition Molecules
journal, January 2007


A Multivalent Marine Lectin from Crenomytilus grayanus Possesses Anti-cancer Activity through Recognizing Globotriose Gb3
journal, March 2016


The genetics and mechanisms of T cell acute lymphoblastic leukaemia
journal, July 2016


Vertebrate protein glycosylation: diversity, synthesis and function
journal, June 2012


Glycosylation in cancer: mechanisms and clinical implications
journal, August 2015


Glycan gene expression signatures in normal and malignant breast tissue; possible role in diagnosis and progression
journal, December 2009


Developing new antibacterials through natural product research
journal, March 2013


Improved Identification and Analysis of Small Open Reading Frame Encoded Polypeptides
journal, March 2016


A General Strategy for the Discovery of Metabolic Pathways: d -Threitol, l -Threitol, and Erythritol Utilization in Mycobacterium smegmatis
journal, November 2015


Galectins: A family of animal β-galactoside-binding lectins
journal, February 1994


Carbohydrate analysis by a phenol–sulfuric acid method in microplate format
journal, April 2005


Expression of LacdiNAc Groups on N-Glycans among Human Tumors Is Complex
journal, January 2014


Specificity of DC-SIGN for mannose- and fucose-containing glycans
journal, October 2006


The small protein floodgates are opening; now the functional analysis begins
journal, December 2014


A new α-galactosyl-binding protein from the mushroom Lyophyllum decastes
journal, November 2007


The Cosmc connection to the Tn antigen in cancer
journal, February 2014


Structural elucidation of a neutral fucogalactan from the mycelium of Coprinus comatus
journal, July 2006


Discovery and characterization of smORF-encoded bioactive polypeptides
journal, November 2015


Bioactive proteins from mushrooms
journal, November 2011


Colorimetric Method for Determination of Sugars and Related Substances
journal, March 1956


Conservation of the C-type lectin fold for massive sequence variation in a Treponema diversity-generating retroelement
journal, August 2011


Biological roles of glycans
journal, August 2016


Chemistry and Biology of Acylfulvenes: Sesquiterpene-Derived Antitumor Agents
journal, March 2012


Mushroom Lectins: Specificity, Structure and Bioactivity Relevant to Human Disease
journal, April 2015


Long non-coding RNAs as a source of new peptides
journal, September 2014


An integrated approach for finding overlooked genes in yeast
journal, January 2002


The Pharmacological Potential of Mushrooms
journal, January 2005


Tissue sulfhydryl groups
journal, May 1959


Long non-coding RNAs as a source of new peptides
text, January 2014


Works referencing / citing this record:

Mechanistic insights into the evolution of DUF26-containing proteins in land plants
journal, February 2019


Activities of two edible macrofungi, Coprinus comatus and Leucoagaricus leucothites in human lymphocytes: cytogenetic and biochemical study
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