Identification of Membrane Proteome of
            <i>Paracoccidioides Lutzii</i>
            and its Regulation by Zinc

Curcio, Juliana Santana de; Silva, Marielle Garcia; Bailão, Mirelle Garcia Silva; Báo, Sônia Nair; Casaletti, Luciana; Bailão, Alexandre Mello; Soares, Célia Maria de Almeida

doi:10.4155/fsoa-2017-0044

Cited by 22 publications

(18 citation statements)

References 99 publications

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“…B8563). Yeast cells of different isolates, cultured in the presence of acetate or glucose, were incubated with aniline blue for 5 min while stirring and subsequently washed twice with 1X PBS (Renshaw et al, 2016 ; De Curcio et al, 2017 ). Samples stained with aniline blue were visualized under a fluorescence microscope (Zeiss Axiocam MRc-Scope A1, Oberkochen, Germany).…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2017

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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.

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Section: Methodsmentioning

confidence: 99%

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

et al. 2017

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“…Antigens of the Paracoccidioides species obtained by immunoprecipitation were subjected to tryptic digestion. Briefly, 10 µl of 50 mM ammonium bicarbonate pH 8.5 were added to 50 µg of the protein extracts and subsequently subjected to tryptic digestion as previously described (Murad and Rech, 2012;Bailao et al, 2014;de Curcio et al, 2017;de Oliveira et al, 2018). Then, 25 µl of 0.2% (w/v) RapiGest SF TM surfactant (Waters Corporation, United Kingdom) was added to the protein extracts and vortexed with subsequent incubation at 80 • C for 15 min.…”

Section: Tryptic Digestion Of Exoantigensmentioning

confidence: 99%

Immunoproteomic Approach of Extracellular Antigens From Paracoccidioides Species Reveals Exclusive B-Cell Epitopes

et al. 2020

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Fungi of the Paracoccidioides genus are the etiological agents of paracoccidioidomycosis (PCM), a systemic mycosis restricted to the countries of Latin America. Currently, the Paracoccidioides complex is represented by Paracoccidioides lutzii, Paracoccidioides americana, Paracoccidioides brasiliensis, Paracoccidioides restrepiensis, and Paracoccidioides venezuelensis. Even with advances in techniques used for diagnosing fungal diseases, high rates of false-positive results for PCM are still presented. Additionally, there is no efficient antigen that can be used to follow up the efficiency of patient treatment. The immunoproteomic is considered a powerful tool for the identification of antigens. In addition, antigens are molecules recognized by the immune system, which make them excellent targets for diagnostic testing of diseases caused by microorganisms. In this vein, we investigated which antigens are secreted by species representing Paracoccidioides complex to increase the spectrum of molecules that could be used for future diagnostic tests, patient follow-up, or PCM therapy. To identify the profile of antigens secreted by Paracoccidioides spp., immunoproteomic approaches were used combining immunoprecipitation, followed by antigen identification by nanoUPLC-MS E-based proteomics. Consequently, it was possible to verify differences in the exoantigen profiles present among the studied species. Through a mass spectrometry approach, it was possible to identify 79 exoantigens in Paracoccidioides species. Using bioinformatics tools, two unique exoantigens in P. lutzii species were identified, as well as 44 epitopes exclusive to the Paracoccidioides complex and 12 unique antigenic sequences that can differentiate between Paracoccidioides species. Therefore, these results demonstrate that Paracoccidioides species have a range of B-cell epitopes exclusive to the complex as well as specific to each Paracoccidioides species. In addition, these analyses allowed us the identification of excellent biomarker candidates for epidemiology screening, diagnosis, patient follow-up, as well as new candidates for PCM therapy.

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“…Standard curves were generated by 1:5 dilution of the cDNA, and the relative expression levels of the transcripts were calculated using the standard curve Analysis of Chitin and Glucan Amount in the Cell Wall of P. brasiliensis During in vitro Copper Deprivation Calcoflour White (CFW, Sigma-Aldrich) and Aniline Blue (AB, Sigma-Aldrich) were used to stain P. brasiliensis yeast cells to evaluate the effect of copper depletion in the composition of the cell wall, since CFW and AB specifically binds chitin and glucan respectively. P. brasiliensis grew in copper depletion or in presence of this metal for 24 h. The cells were stained with the dyes described above and analyzed by fluorescence microscopy as already detailed (De Curcio et al, 2017). In synthesis, for analysis of chitin amount, the cells were fixed in 100% methanol at -80 • C for 20 min.…”

Section: Cdna Synthesis and Quantitative Real Time Pcr (Rt-qpcr) Analmentioning

confidence: 99%

Metabolic Adaptation of Paracoccidioides brasiliensis in Response to in vitro Copper Deprivation

et al. 2020

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Copper is an essential micronutrient for the performance of important biochemical processes such as respiration detoxification, and uptake of metals like iron. Studies have shown that copper deprivation is a strategy used by the host against pathogenic fungi such as Cryptoccocus neoformans and Candida albicans during growth and development of infections in the lungs and kidneys. Although there are some studies, little is known about the impact of copper deprivation in members of the Paracoccidioides genus. Therefore, using isobaric tag labeling (iTRAQ)-Based proteomic approach and LC-MS/MS, we analyzed the impact of in vitro copper deprivation in the metabolism of Paracoccidioides brasiliensis. One hundred and sixty-four (164) differentially abundant proteins were identified when yeast cells were deprived of copper, which affected cellular respiration and detoxification processes. Changes in cellular metabolism such as increased beta oxidation and cell wall remodeling were described.

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Identification of Membrane Proteome of Paracoccidioides Lutzii and its Regulation by Zinc

Cited by 22 publications

References 99 publications

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

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

Immunoproteomic Approach of Extracellular Antigens From Paracoccidioides Species Reveals Exclusive B-Cell Epitopes

Metabolic Adaptation of Paracoccidioides brasiliensis in Response to in vitro Copper Deprivation

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