Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre
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2020-04-27
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Universidade Federal de Goiás
Resumo
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.
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PETITO, G. Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre. 2020.85 f. Tese (Doutorado em Genética e Biologia Molecular) - Universidade Federal de Goiás, Goiânia, 2020.