Proteome of cell wall-extracts from pathogenic Paracoccidioides brasiliensis: Comparison among morphological phases, isolates, and reported fungal extracellular vesicle proteins

• We compared P. brasiliensis pathogenic yeast and mycelial cell wall fraction proteome.
• We compared Pb18 and Pb3 cell wall fraction proteome with focus on yeast phase.
• 92 yeast-exclusive proteins were Pb18 (more virulent)-exclusive.
• 11 yeast-exclusive proteins were Pb3 (less virulent)-exclusive.
• ∼60% of proteins, many cytoplasmic, were described in fungal extracellular vesicles.

We identified non-covalently linked cell wall proteins from Paracoccidioides brasiliensis yeasts and mycelia, with focus on the yeast pathogenic phase, and correlated them with reported fungal extracellular vesicle proteins. We studied isolates Pb3 and Pb18, which evoke distinct patterns of experimental paracoccidioidomycosis and represent two phylogenetic groups. Proteins were extracted mildly with dithiothreitol, trypsinized, and peptides analyzed by liquid chromatography coupled to high-resolution mass spectrometry. Among 132 yeast-exclusive sequences, 92 were Pb18-exclusive. About 80% of total proteins were classified as secretory, mostly showing non-conventional signals. Extracellular vesicular transportation could be involved, since 60% had orthologs reported in fungal extracellular vesicles.

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