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Proteomic analysis of proteome of THP-1 macrophages incubated with Paracoccidioides brasiliensis yeast cells alive or heat-killed

ABSTRACT: Paracoccidioides spp. is the etiological agent of Paracoccidioidomycosis (PCM), a systemicinfection with wide distribution in Latin America. Macrophages are very important cells during the response to infection by Paracoccidoides brasiliensis and understanding the interaction between the fungus and immune cells is very relevant for understanding the disease. In this study, we performed a proteomic analysis to assess the consequences of P. brasiliensis yeast infection on the THP-1 macrophage proteome and to verify whether there are differences between the proteome of cells infected with the live fungus. We identified 443 upregulated and 2247 downregulated proteins in macrophages infected with live P. brasiliensis yeasts, compared to uninfected macrophages. Proteins differentially expressed in cells infected are related to metabolism and energy production, protein synthesis and processing, transcription, cell cycle, DNA processing, cell signaling, oxidative stress, immune response, among other processes . Proteomic analysis revealed that P. brasiliensis, causes metabolic alterations in infected cells, drastically affecting energy production pathways. In addition, macrophages showed many upregulated, mostly downregulated, immune system proteins. Thus, the present work contributes to elucidate the changes that occur in immune cells in response to infection by P. brasiliensis and may help to better understand PCM.

INSTRUMENT(S): Synapt MS

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture

SUBMITTER: Celia Soares

LAB HEAD: Celia Maria Almeida Soares

PROVIDER: PXD043727 | Pride | 2023-10-23

REPOSITORIES: Pride

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			Action	DRS
	20210919_AnaMarina_MeioControle_R1_F1_IA_final_protein.csv	Csv
	20210919_AnaMarina_MeioControle_R2_F1_IA_final_protein.csv	Csv
	20210919_AnaMarina_MeioControle_R3_F1_IA_final_protein.csv	Csv
	20210922_AnaMarina_PbVivo_R1_F1_IA_final_protein.csv	Csv
	20210922_AnaMarina_PbVivo_R2_F1_IA_final_protein.csv	Csv

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Project description:The fungi of Paracoccidioides spp. genus are the causative agents of paracoccidioidomycosis (PCM), a systemic mycosis endemic to Latin America. This fungus is considered a facultative intracellular pathogen that is able to survive and replicate inside macrophages. The survival of the fungus during infection depends on its adaptability to various conditions, such as nitrosative/oxidative stress produced by the host immune cells, particularly alveolar macrophages. Currently, there is little knowledge about the P. brasiliensis signaling pathways involved in the fungus evasion mechanism of the host defense response. However, it is known that some of these pathways are triggered by reactive oxygen species and reactive nitrogen species (ROS/RNS) produced by host cells. Considering that the effects of NO (nitric oxide) on pathogens are concentration dependent, such effects could alter the redox state of cysteine residues by influencing (activating or inhibiting) a variety of protein functions, notably S-nitrosylation, a highly important NO-dependent posttranslational modification that regulates cellular functions and signaling pathways. It has been demonstrated by our group that P. brasiliensis yeast cells proliferate when exposed to low NO concentrations. Thus, this work investigated the modulation profile of S-nitrosylated proteins of P. brasiliensis, as well as identifying S-nitrosylation sites after treatment with RNS. Through mass spectrometry analysis (LC-MS/MS) and label-free quantification, it was possible to identify 474 proteins in the S-nitrosylated proteome study. With this approach, we observed that proteins treated with NO at low concentrations presented a proliferative response pattern, with several proteins involved in cellular cycle regulation and growth being activated. These proteins appear to play important roles in fungal virulence. On the other hand, proteins stimulated by high NO concentrations exhibited a survival response pattern, which may help to elucidate RNS antifungal properties and identify potential molecular targets for the development of new drugs in the future

Dataset Information

Proteomic analysis of proteome of THP-1 macrophages incubated with Paracoccidioides brasiliensis yeast cells alive or heat-killed

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