Abstract
BackgroundThe fungal cell wall is a complex and dynamic outer structure. In pathogenic fungi its components interact with the host, determining the infection fate. The present work aimed to characterize cell wall lipids from P. brasiliensis grown in the presence and absence of human plasma. We compared the results from isolates Pb3 and Pb18, which represent different phylogenetic species that evoke distinct patterns of experimental paracoccidioidomycosis.Methodology/Principal FindingsWe comparatively characterized cell wall phospholipids, fatty acids, sterols, and neutral glycolipids by using both electrospray ionization- and gas chromatography-mass spectrometry analyses of lipids extracted with organic solvents followed by fractionation in silica-gel-60. We detected 49 phospholipid species in Pb3 and 38 in Pb18, including phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, and phosphatidic acid. In both Pb3 and Pb18, PC and PE had the most numerous species. Among the fatty acids, C18∶1 and C18∶2 were the most abundant species in both isolates, although C18∶2 was more abundant in Pb18. There was a different effect of plasma supplementation on fatty acids depending on the fungal isolate. The prevalent glycolipid species was Hex-C18∶0-OH/d19∶2-Cer, although other four minor species were also detected. The most abundant sterol in all samples was brassicasterol. Distinct profiles of cell wall and total yeast sterols suggested that the preparations were enriched for cell wall components. The presence of plasma in the culture medium specially increased cell wall brassicasterol abundance and also other lipids.Conclusions/SignificanceWe here report an original comparative lipidomic analysis of P. brasiliensis cell wall. Our results open doors to understanding the role of cell wall lipids in fungal biology, and interaction with anti-fungal drugs and the host.
Highlights
The cell wall is a complex and dynamic fungal structure involved in functions such as maintenance of the cell shape, protection from mechanical injuries, adaptation to morphogenesis, and withstanding osmotic pressure [1]
We compared the results for Pb3 and Pb18 cultivated in the presence or absence of human plasma
Phospholipids from Pb3 and Pb18 cell wall preparations were ionized in positive- (Fig. 1A) and negative-ion (Fig. 1B) modes of electrospray ionization- (ESI-)MS/MS
Summary
The cell wall is a complex and dynamic fungal structure involved in functions such as maintenance of the cell shape, protection from mechanical injuries, adaptation to morphogenesis, and withstanding osmotic pressure [1]. It is the main structure in contact with the host, directly participating in hostfungal relationship [2]. Dimorphic fungi constitute a small group of pathogens that develop as yeast or mycelia according to the temperature of growth [4] In this group, the cell wall lipid content varies with the fungal species and/or morphological phase, ranging from 5.5%, in Blastomyces dermatitidis pathogenic yeasts, to 26% in Sporothrix schenckii environmental hyphae [5]. We compared the results from isolates Pb3 and Pb18, which represent different phylogenetic species that evoke distinct patterns of experimental paracoccidioidomycosis
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