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Title: Extracellular vesicles from Paracoccidioides pathogenic species transport polysaccharide and expose ligands for DC-SIGN receptors

Abstract

Extracellular vesicles (EVs) mediate non-conventional transport of molecules across the fungal cell wall. We aimed at describing the carbohydrate composition and surface carbohydrate epitopes of EVs isolated from the pathogenic fungi Paracoccidioides brasiliensis and P. lutzii using standard procedures. Total EV carbohydrates were ethanol-precipitated from preparations depleted of lipids and proteins, then analyzed by chemical degradation, gas chromatography-mass spectrometry, nuclear magnetic resonance and size-exclusion chromatography. EV glycosyl residues of Glc, Man, and Gal comprised most probably two major components: a high molecular mass 4,6-α-glucan and a galactofuranosylmannan, possibly an oligomer, bearing a 2-α-Manp main chain linked to β-Galf (1,3) and α-Manp (1,6) end units. The results also suggested the presence of small amounts of a (1→6)- Manp polymer, (1→3)-glucan and (1→6)-glucan. Glycan microarrays allowed identification of EV surface lectin(s), while plant lectin microarray profiling revealed terminal Man and GlcNAc residues exposed at the EVs surface. Mammalian lectin microarray profiling showed that DC-SIGN receptors recognized surface carbohydrate in Paracoccidioides EVs. Our results suggest that oligosaccharides, cytoplasmic storage, and cell wall polysaccharides can be exported in fungal EVs, which also expose surface PAMPs and lectins. As a result, the role of these newly identified components in the interaction with the host remainsmore » to be unraveled.« less

Authors:
 [1];  [2];  [2];  [2];  [3];  [1];  [3];  [4];  [1]
  1. Escola Paulista de Medicina-Univ. Federal de Sao Paulo, Sao Paulo (Brazil)
  2. Univ. of Georgia, Athens, GA (United States)
  3. National Centre for Biomedical Engineering Science, NUI Galway (Ireland)
  4. National Centre for Biomedical Engineering Science, NUI Galway (Ireland); National Univ. of Ireland, Galway (Ireland)
Publication Date:
Research Org.:
Univ. of Georgia, Athens, GA (United States). Analytical Services & Training Lab. Complex Carbohydrate Research Center
Sponsoring Org.:
USDOE
OSTI Identifier:
1324971
Grant/Contract Number:  
FG02-93ER20097
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 5; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; fungal biology; glycoconjugates; membrane trafficking; polysaccharides

Citation Formats

da Silva, Roberta Peres, Heiss, Christian, Black, Ian, Azadi, Parastoo, Gerlach, Jared Q., Travassos, Luiz R., Joshi, Lokesh, Kilcoyne, Michelle, and Puccia, Rosana. Extracellular vesicles from Paracoccidioides pathogenic species transport polysaccharide and expose ligands for DC-SIGN receptors. United States: N. p., 2015. Web. doi:10.1038/srep14213.
da Silva, Roberta Peres, Heiss, Christian, Black, Ian, Azadi, Parastoo, Gerlach, Jared Q., Travassos, Luiz R., Joshi, Lokesh, Kilcoyne, Michelle, & Puccia, Rosana. Extracellular vesicles from Paracoccidioides pathogenic species transport polysaccharide and expose ligands for DC-SIGN receptors. United States. doi:10.1038/srep14213.
da Silva, Roberta Peres, Heiss, Christian, Black, Ian, Azadi, Parastoo, Gerlach, Jared Q., Travassos, Luiz R., Joshi, Lokesh, Kilcoyne, Michelle, and Puccia, Rosana. Mon . "Extracellular vesicles from Paracoccidioides pathogenic species transport polysaccharide and expose ligands for DC-SIGN receptors". United States. doi:10.1038/srep14213. https://www.osti.gov/servlets/purl/1324971.
@article{osti_1324971,
title = {Extracellular vesicles from Paracoccidioides pathogenic species transport polysaccharide and expose ligands for DC-SIGN receptors},
author = {da Silva, Roberta Peres and Heiss, Christian and Black, Ian and Azadi, Parastoo and Gerlach, Jared Q. and Travassos, Luiz R. and Joshi, Lokesh and Kilcoyne, Michelle and Puccia, Rosana},
abstractNote = {Extracellular vesicles (EVs) mediate non-conventional transport of molecules across the fungal cell wall. We aimed at describing the carbohydrate composition and surface carbohydrate epitopes of EVs isolated from the pathogenic fungi Paracoccidioides brasiliensis and P. lutzii using standard procedures. Total EV carbohydrates were ethanol-precipitated from preparations depleted of lipids and proteins, then analyzed by chemical degradation, gas chromatography-mass spectrometry, nuclear magnetic resonance and size-exclusion chromatography. EV glycosyl residues of Glc, Man, and Gal comprised most probably two major components: a high molecular mass 4,6-α-glucan and a galactofuranosylmannan, possibly an oligomer, bearing a 2-α-Manp main chain linked to β-Galf (1,3) and α-Manp (1,6) end units. The results also suggested the presence of small amounts of a (1→6)- Manp polymer, (1→3)-glucan and (1→6)-glucan. Glycan microarrays allowed identification of EV surface lectin(s), while plant lectin microarray profiling revealed terminal Man and GlcNAc residues exposed at the EVs surface. Mammalian lectin microarray profiling showed that DC-SIGN receptors recognized surface carbohydrate in Paracoccidioides EVs. Our results suggest that oligosaccharides, cytoplasmic storage, and cell wall polysaccharides can be exported in fungal EVs, which also expose surface PAMPs and lectins. As a result, the role of these newly identified components in the interaction with the host remains to be unraveled.},
doi = {10.1038/srep14213},
journal = {Scientific Reports},
number = ,
volume = 5,
place = {United States},
year = {2015},
month = {9}
}

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