Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre
Autor(a) principal: | |
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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 |
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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Soares, Célia Maria de Almeidahttp://lattes.cnpq.br/8539946335852637Soares, Célia Maria de AlmeidaCastro, Agenor deSilva, Lívia do CarmoCurcio, Juliana Santana deSilva, Kleber Santiago Freitas ehttp://lattes.cnpq.br/9484967397025799Petito, Guilherme2020-09-10T15:39:26Z2020-09-10T15:39:26Z2020-04-27PETITO, 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.http://repositorio.bc.ufg.br/tede/handle/tede/10648Copper 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.O cobre é um importante micronutriente que atua como cofator em enzimas que participam em processos de obtenção de energia, redução de espécies reativas de oxigênio e captação de metal, como o ferro. Estudos com diferentes fungos patogênicos demonstram a importância de um sistema eficiente de homeostase de cobre, bem como o impacto da privação desse metal no metabolismo desses organismos. Entretanto, pouco se sabe acerca do impacto da privação de cobre em fungos patogênicos do gênero Paracoccidioides. Dessa forma, objetivou-se com este estudo analisar o comportamento proteômico do fungo Paracoccidioides brasiliensis durante a privação de cobre, identificando as alterações metabólicas frente a essa condição. A análise proteômicas foi realizada utilizando espectrometria de massas e marcação das amostras com iTRAQ (Isobariq tag for relative and absolute quantitation). Além disso, foram aplicadas neste estudo metodologias como qRTPCR, microscopia de fluorescência, atividades enzimáticas e Western bloting, com intuito de validar os dados proteômicos analisados. Foram identificadas 183 proteínas diferencialmente expressas. Proteínas relacionadas à gliconeogênese, beta-oxidação e remodelamento de parede estavam com expressão aumentada na condição de privação de cobre. Identificamos diminuição na expressão de uma importante enzima de detoxificação, a superóxido dismutase Cu/Zn dependente (Sod1p). Por outro lado, as enzimas tioredoxina (Trxp), glutationa-S-transferase (GST) e superóxido dismutase Mn/Fe dependente (Sod2p) estavam up-reguladas. Também enzimas da via das pentoses-fosfato relacionadas à produção de NADPH, importante agente redudor de enzimas de detoxificação foram aumentadas. Uma oxidase alternativa (Aoxp) estava com expressão aumentada, enquanto testes de atividades enzimáticas demonstraram que a atividade da enzima citocromo c oxidase (Coxp) estava diminuída. A expressão chaperona de cobre, Atx1, relacionada com captação de ferro e descrita como importante em reposta ao estresse oxidativo, estava aumentada. Propomos aqui uma visão geral que apresenta as principais mudanças metabólicas em P. brasiliensis sob privação de cobre, que incluem mudanças no processo de obtenção de energia, no fluxo do metabolismo de carbono, no remodelamento de parede, nos processos de detoxificação e de respiração.Submitted by Liliane Ferreira (ljuvencia30@gmail.com) on 2020-09-10T13:54:05Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Tese - Guilherme Petito - 2020.pdf: 28057865 bytes, checksum: 36c41883059eb468d06d62202e921074 (MD5)Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2020-09-10T15:39:25Z (GMT) No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Tese - Guilherme Petito - 2020.pdf: 28057865 bytes, checksum: 36c41883059eb468d06d62202e921074 (MD5)Made available in DSpace on 2020-09-10T15:39:26Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Tese - Guilherme Petito - 2020.pdf: 28057865 bytes, checksum: 36c41883059eb468d06d62202e921074 (MD5) Previous issue date: 2020-04-27Fundação de Amparo à Pesquisa do Estado de GoiásporUniversidade Federal de GoiásPrograma de Pós-graduação em Genética e Biologia Molecular (ICB)UFGBrasilInstituto de Ciências Biológicas - ICB (RG)Attribution-NonCommercial-NoDerivs 3.0 Brazilhttp://creativecommons.org/licenses/by-nc-nd/3.0/br/info:eu-repo/semantics/openAccessParacoccidioides spp.ProteomicaiTRAQParacoccidiodomicoseProteomicParacoccidioidomycosisCIENCIAS BIOLOGICASAnálise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobreProteomic analysis of the fungus Paracoccidioides brasiliensis under copper deprivationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesis55500500500500241723reponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.bc.ufg.br/tede/bitstreams/895dcfc5-70ad-4978-a4a2-aa28e89a2edd/download8a4605be74aa9ea9d79846c1fba20a33MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8811http://repositorio.bc.ufg.br/tede/bitstreams/da80b446-045b-49ca-a90b-281c5a903614/downloade39d27027a6cc9cb039ad269a5db8e34MD52ORIGINALTese - Guilherme Petito - 2020.pdfTese - Guilherme Petito - 2020.pdfapplication/pdf28057865http://repositorio.bc.ufg.br/tede/bitstreams/fe4daa16-8f15-49e7-b41f-444b0e88cc19/download36c41883059eb468d06d62202e921074MD53tede/106482020-09-10 12:39:29.495http://creativecommons.org/licenses/by-nc-nd/3.0/br/Attribution-NonCommercial-NoDerivs 3.0 Brazilopen.accessoai:repositorio.bc.ufg.br:tede/10648http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2020-09-10T15:39:29Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)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 |
dc.title.pt_BR.fl_str_mv |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
dc.title.alternative.eng.fl_str_mv |
Proteomic analysis of the fungus Paracoccidioides brasiliensis under copper deprivation |
title |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
spellingShingle |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre Petito, Guilherme Paracoccidioides spp. Proteomica iTRAQ Paracoccidiodomicose Proteomic Paracoccidioidomycosis CIENCIAS BIOLOGICAS |
title_short |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
title_full |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
title_fullStr |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
title_full_unstemmed |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
title_sort |
Análise proteômica do fungo Paracoccidioides brasiliensis sob privação de cobre |
author |
Petito, Guilherme |
author_facet |
Petito, Guilherme |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
Soares, Célia Maria de Almeida |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/8539946335852637 |
dc.contributor.referee1.fl_str_mv |
Soares, Célia Maria de Almeida |
dc.contributor.referee2.fl_str_mv |
Castro, Agenor de |
dc.contributor.referee3.fl_str_mv |
Silva, Lívia do Carmo |
dc.contributor.referee4.fl_str_mv |
Curcio, Juliana Santana de |
dc.contributor.referee5.fl_str_mv |
Silva, Kleber Santiago Freitas e |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/9484967397025799 |
dc.contributor.author.fl_str_mv |
Petito, Guilherme |
contributor_str_mv |
Soares, Célia Maria de Almeida Soares, Célia Maria de Almeida Castro, Agenor de Silva, Lívia do Carmo Curcio, Juliana Santana de Silva, Kleber Santiago Freitas e |
dc.subject.por.fl_str_mv |
Paracoccidioides spp. Proteomica iTRAQ Paracoccidiodomicose |
topic |
Paracoccidioides spp. Proteomica iTRAQ Paracoccidiodomicose Proteomic Paracoccidioidomycosis CIENCIAS BIOLOGICAS |
dc.subject.eng.fl_str_mv |
Proteomic Paracoccidioidomycosis |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS |
description |
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. |
publishDate |
2020 |
dc.date.accessioned.fl_str_mv |
2020-09-10T15:39:26Z |
dc.date.available.fl_str_mv |
2020-09-10T15:39:26Z |
dc.date.issued.fl_str_mv |
2020-04-27 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
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. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tede/10648 |
identifier_str_mv |
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. |
url |
http://repositorio.bc.ufg.br/tede/handle/tede/10648 |
dc.language.iso.fl_str_mv |
por |
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por |
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500 500 500 500 |
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Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Attribution-NonCommercial-NoDerivs 3.0 Brazil http://creativecommons.org/licenses/by-nc-nd/3.0/br/ |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.publisher.program.fl_str_mv |
Programa de Pós-graduação em Genética e Biologia Molecular (ICB) |
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UFG |
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Brasil |
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Instituto de Ciências Biológicas - ICB (RG) |
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Universidade Federal de Goiás |
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