来自副球孢子菌致病物种的细胞外囊泡运输多糖并暴露DC-SIGN受体的配体。

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

作者信息

Peres da Silva Roberta, Heiss Christian, Black Ian, Azadi Parastoo, Gerlach Jared Q, Travassos Luiz R, Joshi Lokesh, Kilcoyne Michelle, Puccia Rosana

机构信息

Escola Paulista de Medicina-Universidade Federal de São Paulo, São Paulo, Brazil.

Analytical Services &Training Laboratory Complex Carbohydrate Research Center, The University of Georgia.

出版信息

Sci Rep. 2015 Sep 21;5:14213. doi: 10.1038/srep14213.

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. The role of these newly identified components in the interaction with the host remains to be unraveled.

摘要

细胞外囊泡(EVs)介导分子跨真菌细胞壁的非常规运输。我们旨在使用标准程序描述从致病真菌巴西副球孢子菌和卢氏副球孢子菌中分离出的EVs的碳水化合物组成和表面碳水化合物表位。从去除脂质和蛋白质的制剂中乙醇沉淀总EV碳水化合物,然后通过化学降解、气相色谱-质谱、核磁共振和尺寸排阻色谱进行分析。Glc、Man和Gal的EV糖基残基最有可能包括两个主要成分:一种高分子量的4,6-α-葡聚糖和一种半乳呋喃糖基甘露聚糖,可能是一种寡聚物,带有与β-Galf(1,3)和α-Manp(1,6)末端单元相连的2-α-Manp主链。结果还表明存在少量的(1→6)-Manp聚合物、(1→3)-葡聚糖和(1→6)-葡聚糖。聚糖微阵列可鉴定EV表面凝集素,而植物凝集素微阵列分析揭示了暴露于EV表面的末端Man和GlcNAc残基。哺乳动物凝集素微阵列分析表明,DC-SIGN受体识别副球孢子菌EVs表面的碳水化合物。我们的结果表明,寡糖、细胞质储存物和细胞壁多糖可通过真菌EVs输出,EVs还暴露表面病原体相关分子模式(PAMPs)和凝集素。这些新鉴定成分在与宿主相互作用中的作用仍有待阐明。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/789c/4585699/1f8fd0c73900/srep14213-f1.jpg

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