Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre

• Autor(a) principal: Petito, Guilherme
• Data de Publicação: 2020
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Repositório Institucional da UFG
• Texto Completo: http://repositorio.bc.ufg.br/tede/handle/tede/10648
• topic: Paracoccidioides spp.; Proteomica; iTRAQ; Paracoccidiodomicose; Proteomic; Paracoccidioidomycosis; CIENCIAS BIOLOGICAS

Copper is an important micronutrient that acts as a cofactor in important enzymes that participate in processes of obtaining energy, reducing reactive oxygen species and metal uptake such as iron. Studies with different pathogenic fungi demonstrate the importance of an efficient copper homeostasis system, as well as the impact of this metal deprivation on the metabolism of these organisms. However, little is known about the impact of copper deprivation on pathogenic fungi of the genus Paracoccidioides. Thus, the objective of this study was to analyze the proteomic behavior of the fungus Paracoccidioides brasiliensis during copper deprivation, identifying metabolic changes in face of this condition. Proteomic analysis was performed using mass spectrometry and labelling the samples with iTRAQ (Isobariq tag for relative and absolute quantitation). In addition, methodologies such as qRT-PCR, fluorescence microscopy, enzymatic activities and Western bloting were applied in this study in order to support the proteomic data analyzed. We identified 183 differentially expressed proteins. Proteins related to gluconeogenesis, beta-oxidation and cell wall remodeling were up-regulated. We identified decreased expression of an important detoxification enzyme, Cu / Zn dependent Superoxide Dismutase (Sod1p). On the other hand, the enzymes Thioredoxin (Trxp), Glutathione S-transferase (GST) and Mn / Fedependent superoxide dismutase (Sod2p) were up-regulated. Also, NADPH production-related to the pentose-phosphate pathway enzymes. detoxifying enzyme reducing agent. An alternative oxidase (Aoxp) was up-regulated, while enzyme activity tests showed that the activity of the enzyme Cytochrome c oxidase (Coxp) was decreased. An iron uptake-related copper chaperone, Atx1 and described as important in response to oxidative stress, was up-regulated. We propose here an overview that presents the main metabolic changes in P. brasiliensis under copper deprivation and that include changes in the energy acquisition process, carbon metabolism flow, wall modeling, detoxification and respiration processes.