Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis
Autor(a) principal: | |
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Data de Publicação: | 2009 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFG |
Texto Completo: | http://repositorio.bc.ufg.br/tede/handle/tde/1793 |
Resumo: | The pathogenic fungus Paracoccidioides brasiliensis is the etiologic agent of Paracoccidioidomycosis (PCM). The main route of contamination is the inhalation of fungal propagules that convert to yeast in the host tissues. The urease is among the factors of virulence described for dimorphic fungus. It hydrolyzes the urea producing molecules of ammonia and carbamate. Pbure has 3,012 bp, corresponding to a predicted protein of 837 amino acids, predicted molecular mass of 90 kDa and pI 6.0. PbURE has signature to nickel-dependent enzyme for its activity. The phylogenetic relationship between PbURE and urease from other fungi was evaluated. The cDNA that encodes PbURE was inserted into the expression vector pET-32a and recombinant protein of 103 kDa was expressed. Polyclonal antibody were produced against PbUREr and used on Western blot, Far-Western blot and ELISA. The results showed that PbUREr interferes on interaction between P. brasiliensis and pulmonary epithelial cells A549. PbUREr was able to bind to proteins fibronectin and type IV collagen, components of the extracellular matrix (ECM). The interaction between PbURE and calnexin protein was identified by the technique of two-hybrid. |
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PEREIRA, Maristelahttp://lattes.cnpq.br/1345781867765758http://lattes.cnpq.br/9221913447597085FERNANDES, Maria Regilda de Araújo2014-07-29T15:30:35Z2011-02-282009-02-27FERNANDES, Maria Regilda de Araújo. Molecular and Functional Characterization of Urease of P. brasiliensis. 2009. 68 f. Dissertação (Mestrado em Medicina) - Universidade Federal de Goiás, Goiânia, 2009.http://repositorio.bc.ufg.br/tede/handle/tde/1793The pathogenic fungus Paracoccidioides brasiliensis is the etiologic agent of Paracoccidioidomycosis (PCM). The main route of contamination is the inhalation of fungal propagules that convert to yeast in the host tissues. The urease is among the factors of virulence described for dimorphic fungus. It hydrolyzes the urea producing molecules of ammonia and carbamate. Pbure has 3,012 bp, corresponding to a predicted protein of 837 amino acids, predicted molecular mass of 90 kDa and pI 6.0. PbURE has signature to nickel-dependent enzyme for its activity. The phylogenetic relationship between PbURE and urease from other fungi was evaluated. The cDNA that encodes PbURE was inserted into the expression vector pET-32a and recombinant protein of 103 kDa was expressed. Polyclonal antibody were produced against PbUREr and used on Western blot, Far-Western blot and ELISA. The results showed that PbUREr interferes on interaction between P. brasiliensis and pulmonary epithelial cells A549. PbUREr was able to bind to proteins fibronectin and type IV collagen, components of the extracellular matrix (ECM). The interaction between PbURE and calnexin protein was identified by the technique of two-hybrid.O fungo patogênico humano Paracoccidioides brasiliensis é o agente etiológico da Paracoccidioidomicose (PCM). A principal via de contaminação é a inalação de micélios ou conídios do fungo que se convertem em leveduras no hospedeiro. Dentre os fatores de virulência descritos para fungos dimórficos está a urease, a qual hidrolisa uréia produzindo amônia e carbamato. Pbure possui 3.012 pb, correspondendo a uma proteína predita de 837 aminoácidos, massa molecular predita de 90 kDa e pI 6.0. PbURE possui assinatura de uma enzima dependente de níquel para sua atividade. A relação filogenética entre PbURE e ureases de outros fungos foi avaliada. O cDNA que codifica PbURE foi inserido no vetor de expressão pET-32a e a proteína recombinante de 103 kDa foi expressa. Anticorpo policlonal foi produzido contra PbUREr e utilizados nos ensaios de Western blot, Far-Western blot e ELISA. Os resultados mostram que PbUREr interfere na interação entre P. brasiliensis e células epiteliais pulmonares A549. PbUREr foi capaz de se ligar às proteínas fibronectina e colágeno tipo IV, componentes da matriz extracelular (MEC). A interação entre PbURE e a proteína calnexina foi identificada através da técnica de duplo-híbrido.Made available in DSpace on 2014-07-29T15:30:35Z (GMT). No. of bitstreams: 1 dissertacao maria regilda araujo fernandes.pdf: 889828 bytes, checksum: eeac21e7e2102f6042640b9c030c3870 (MD5) Previous issue date: 2009-02-27application/pdfhttp://repositorio.bc.ufg.br/TEDE/retrieve/4666/dissertacao%20maria%20regilda%20araujo%20fernandes.pdf.jpgporUniversidade Federal de GoiásMestrado em Medicina TropicalUFGBRMedicinaParacoccidioides brasiliensisUreaseRelações filogenéticasExpressão heterólogaInterações intermolecularesParacoccidioides brasiliensisUreasePhylogenetic relationshipsHeterologous expressionIntermolecular InteractionsCNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIACaracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensisMolecular and Functional Characterization of Urease of P. brasiliensisinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFGinstname:Universidade Federal de Goiás (UFG)instacron:UFGORIGINALdissertacao maria regilda araujo fernandes.pdfapplication/pdf889828http://repositorio.bc.ufg.br/tede/bitstreams/d7db8493-2fd5-4dab-863f-ac654876eecc/downloadeeac21e7e2102f6042640b9c030c3870MD51THUMBNAILdissertacao maria regilda araujo fernandes.pdf.jpgdissertacao maria regilda araujo fernandes.pdf.jpgGenerated Thumbnailimage/jpeg3962http://repositorio.bc.ufg.br/tede/bitstreams/c5c2eddd-f582-4e08-ad7b-65338e1480fe/download2e5b6e7718ce886421aa7755530fb7d8MD52tde/17932014-07-30 03:15:41.235open.accessoai:repositorio.bc.ufg.br:tde/1793http://repositorio.bc.ufg.br/tedeRepositório InstitucionalPUBhttp://repositorio.bc.ufg.br/oai/requesttasesdissertacoes.bc@ufg.bropendoar:2014-07-30T06:15:41Repositório Institucional da UFG - Universidade Federal de Goiás (UFG)false |
dc.title.por.fl_str_mv |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
dc.title.alternative.eng.fl_str_mv |
Molecular and Functional Characterization of Urease of P. brasiliensis |
title |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
spellingShingle |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis FERNANDES, Maria Regilda de Araújo Paracoccidioides brasiliensis Urease Relações filogenéticas Expressão heteróloga Interações intermoleculares Paracoccidioides brasiliensis Urease Phylogenetic relationships Heterologous expression Intermolecular Interactions CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
title_short |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
title_full |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
title_fullStr |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
title_full_unstemmed |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
title_sort |
Caracterização Molecular e Funcional da Urease de Paracoccidioides brasiliensis |
author |
FERNANDES, Maria Regilda de Araújo |
author_facet |
FERNANDES, Maria Regilda de Araújo |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
PEREIRA, Maristela |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/1345781867765758 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/9221913447597085 |
dc.contributor.author.fl_str_mv |
FERNANDES, Maria Regilda de Araújo |
contributor_str_mv |
PEREIRA, Maristela |
dc.subject.por.fl_str_mv |
Paracoccidioides brasiliensis Urease Relações filogenéticas Expressão heteróloga Interações intermoleculares |
topic |
Paracoccidioides brasiliensis Urease Relações filogenéticas Expressão heteróloga Interações intermoleculares Paracoccidioides brasiliensis Urease Phylogenetic relationships Heterologous expression Intermolecular Interactions CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
dc.subject.eng.fl_str_mv |
Paracoccidioides brasiliensis Urease Phylogenetic relationships Heterologous expression Intermolecular Interactions |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::MICROBIOLOGIA |
description |
The pathogenic fungus Paracoccidioides brasiliensis is the etiologic agent of Paracoccidioidomycosis (PCM). The main route of contamination is the inhalation of fungal propagules that convert to yeast in the host tissues. The urease is among the factors of virulence described for dimorphic fungus. It hydrolyzes the urea producing molecules of ammonia and carbamate. Pbure has 3,012 bp, corresponding to a predicted protein of 837 amino acids, predicted molecular mass of 90 kDa and pI 6.0. PbURE has signature to nickel-dependent enzyme for its activity. The phylogenetic relationship between PbURE and urease from other fungi was evaluated. The cDNA that encodes PbURE was inserted into the expression vector pET-32a and recombinant protein of 103 kDa was expressed. Polyclonal antibody were produced against PbUREr and used on Western blot, Far-Western blot and ELISA. The results showed that PbUREr interferes on interaction between P. brasiliensis and pulmonary epithelial cells A549. PbUREr was able to bind to proteins fibronectin and type IV collagen, components of the extracellular matrix (ECM). The interaction between PbURE and calnexin protein was identified by the technique of two-hybrid. |
publishDate |
2009 |
dc.date.issued.fl_str_mv |
2009-02-27 |
dc.date.available.fl_str_mv |
2011-02-28 |
dc.date.accessioned.fl_str_mv |
2014-07-29T15:30:35Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
FERNANDES, Maria Regilda de Araújo. Molecular and Functional Characterization of Urease of P. brasiliensis. 2009. 68 f. Dissertação (Mestrado em Medicina) - Universidade Federal de Goiás, Goiânia, 2009. |
dc.identifier.uri.fl_str_mv |
http://repositorio.bc.ufg.br/tede/handle/tde/1793 |
identifier_str_mv |
FERNANDES, Maria Regilda de Araújo. Molecular and Functional Characterization of Urease of P. brasiliensis. 2009. 68 f. Dissertação (Mestrado em Medicina) - Universidade Federal de Goiás, Goiânia, 2009. |
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http://repositorio.bc.ufg.br/tede/handle/tde/1793 |
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Universidade Federal de Goiás |
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Mestrado em Medicina Tropical |
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UFG |
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BR |
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Medicina |
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Universidade Federal de Goiás |
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