Differential Metabolism of a Two-Carbon Substrate by Members of the Paracoccidioides Genus

Abstract

The genus Paracoccidioides comprises known fungal pathogens of humans and can be isolated from different infection sites. Metabolic peculiarities in different members of the Paracoccidioides led us to perform proteomic studies in the presence of the two-carbon molecule acetate, which predominates in the nutrient-poor environment of the phagosome. To investigate the expression rates of proteins of different members of Paracoccidioides, including one isolate of P. lutzii (Pb01) and three isolates of P. brasiliensis (Pb03, Pb339, and PbEPM83), using sodium acetate as a carbon source, proteins were quantified using label-free and data-independent liquid chromatography-mass spectrometry. Protein profiles of the isolates were statistically analyzed, revealing proteins that were differentially expressed when the fungus was cultivated in a non-preferential carbon source rather than glucose. A total of 1,160, 1,211, 1,280, and 1,462 proteins were reproducibly identified and relatively quantified in P. lutzii and the P. brasiliensis isolates Pb03, Pb339, and PbEPM83, respectively. Notably, 526, 435, 744, and 747 proteins were differentially expressed among P. lutzii and the P. brasiliensis isolates Pb03, Pb339, and PbEPM83, respectively, with a fold-change equal to or higher than 1.5. This analysis revealed that reorganization of metabolism occurred through the induction of proteins related to gluconeogenesis, glyoxylic/glyoxylate cycle, response to stress, and degradation of amino acids in the four isolates. The following differences were observed among the isolates: higher increases in the expression levels of proteins belonging to the TCA and respiratory chain in PbEPM83 and Pb01; increase in ethanol production in Pb01; utilization of cell wall components for gluconeogenesis in Pb03 and PbEPM83; and increased β-oxidation and methylcitrate cycle proteins in Pb01and PbEPM83. Proteomic profiles indicated that the four isolates reorganized their metabolism in different manners to use acetate as a carbon source.

Molecular characterization of siderophore biosynthesis in Paracoccidioides brasiliensis

“…Protein extraction was performed by protocol described by Baeza and coleagues (Baeza et al 2017 ). The protein concentration in the supernatant was determined using the Bradford reagent (Bradford 1976 ).…”

Insights Into Histoplasma capsulatum Behavior on Zinc Deprivation

“…The samples were centrifuged, and supernatant was lyophilized in the speed vacuum (Eppendorf, Hamburg, Germany). The tryptic peptides were separated by Ultra High Performance Liquid Chromatography according to Baeza et al (2017), using the ACQUITY UPLC ® M-Class system (Waters Corporation, Manchester, UK) coupled to Synapt G1 HDMS ™ mass spectrometer (Waters Micromass, Manchester, UK). Rabbit phosphorylase B (MassPREP ™ Digestion Standard) was used as internal standard and [GLU1]-Fibrinopeptide B (GFB) for calibration during the sample analysis.…”

Metabolic Peculiarities of Paracoccidioides brasiliensis Dimorphism as Demonstrated by iTRAQ Labeling Proteomics

“…When yeast cells were grown in acetate as a carbon source, the metabolic profile depicted gluconeogenesis and catabolism of amino acids induced at similar rates among members of the Paracoccidioides genus. For P. lutzii and P. restrepiensis , β-oxidation, TCA, and glyoxylate cycles were increased in comparison to P. brasiliensis and P. americana (Baeza et al, 2017). The data cited above reinforce the proposition of metabolic differences in members of the Paracoccidioide s genus.…”

“…Metabolic differences are described in members of the Paracoccidioide s genus that could account to their adaptation to the host (Pigosso et al, 2013; Baeza et al, 2017; Lacerda Pigosso et al, 2017). Previous work from our group evidenced that when yeast cells have glucose as the carbon source, proteins related to glycolysis/gluconeogenesis and to alcohol fermentation were more accumulated in P. lutzii when compared to other members of the genus.…”