Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1093/femsyr/fow079 http://hdl.handle.net/11449/173769 |
Resumo: | Paracoccidioides spp., which are temperature-dependent dimorphic fungi, are responsible for the most prevalent human systemic mycosis in Latin America, the paracoccidioidomycosis. The aim of this study was to characterise the involvement of elongation factor Tu (EF-Tu) in Paracoccidioides brasiliensis-host interaction. Adhesive properties were examined using recombinant PbEF-Tu proteins and the respective polyclonal anti-rPbEF-Tu antibody. Immunogold analysis demonstrated the surface location of EF-Tu in P. brasiliensis. Moreover, PbEF-Tu was found to bind to fibronectin and plasminogen by enzyme-linked immunosorbent assay, and it was determined that the binding to plasminogen is at least partly dependent on lysine residues and ionic interactions. To verify the participation of EF-Tu in the interaction of P. brasiliensis with pneumocytes, we blocked the respective protein with an anti-rPbEF-Tu antibody and evaluated the consequences on the interaction index by flow cytometry. During the interaction, we observed a decrease of 2- and 3-fold at 8 and 24 h, respectively, suggesting the contribution of EF-Tu in fungal adhesion/invasion. |
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Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interactionEF-Tufibronectinfungal-host interactionmoonlighting proteinParacoccidioides brasiliensisplasminogenParacoccidioides spp., which are temperature-dependent dimorphic fungi, are responsible for the most prevalent human systemic mycosis in Latin America, the paracoccidioidomycosis. The aim of this study was to characterise the involvement of elongation factor Tu (EF-Tu) in Paracoccidioides brasiliensis-host interaction. Adhesive properties were examined using recombinant PbEF-Tu proteins and the respective polyclonal anti-rPbEF-Tu antibody. Immunogold analysis demonstrated the surface location of EF-Tu in P. brasiliensis. Moreover, PbEF-Tu was found to bind to fibronectin and plasminogen by enzyme-linked immunosorbent assay, and it was determined that the binding to plasminogen is at least partly dependent on lysine residues and ionic interactions. To verify the participation of EF-Tu in the interaction of P. brasiliensis with pneumocytes, we blocked the respective protein with an anti-rPbEF-Tu antibody and evaluated the consequences on the interaction index by flow cytometry. During the interaction, we observed a decrease of 2- and 3-fold at 8 and 24 h, respectively, suggesting the contribution of EF-Tu in fungal adhesion/invasion.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Faculdade de Ciências Farmacêuticas UNESP - Univ Estadual Paulista Departamento de Análises Clínicas Laboratório de Micologia ClínicaInstituto de Química UNESP - Univ Estadual Paulista Departamento de Bioquímica e Tecnologia Química Unidade de Sintese Estrutura e Caracterizacao de Peptideos e ProteinasFaculdade de Ciências Farmacéuticas UNESP - Univ Estadual Paulista Departamento de Análises Clínicas Laboratório de Imunologia ClínicaFaculdade de Ciências Farmacêuticas UNESP - Univ Estadual Paulista Departamento de Análises Clínicas Laboratório de Micologia ClínicaInstituto de Química UNESP - Univ Estadual Paulista Departamento de Bioquímica e Tecnologia Química Unidade de Sintese Estrutura e Caracterizacao de Peptideos e ProteinasFaculdade de Ciências Farmacéuticas UNESP - Univ Estadual Paulista Departamento de Análises Clínicas Laboratório de Imunologia ClínicaFAPESP: 2015/03700-9Universidade Estadual Paulista (Unesp)Marcos, Caroline Maria [UNESP]de Oliveira, Haroldo Cesar [UNESP]da Silva, Julhiany de Fátima [UNESP]Assato, Patricia Akemi [UNESP]Yamazaki, Daniella Sayuri [UNESP]da Silva, Rosângela Aparecida Moraes [UNESP]Santos, Cláudia Tavares [UNESP]Santos-Filho, Norival Alves [UNESP]Portuondo, Deivys Leandro [UNESP]Mendes-Giannini, Maria José Soares [UNESP]Fusco-Almeida, Ana Marisa [UNESP]2018-12-11T17:07:39Z2018-12-11T17:07:39Z2016-11-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1093/femsyr/fow079FEMS Yeast Research, v. 16, n. 7, 2016.1567-13641567-1356http://hdl.handle.net/11449/17376910.1093/femsyr/fow0792-s2.0-849947179652-s2.0-84994717965.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengFEMS Yeast Research1,3081,308info:eu-repo/semantics/openAccess2024-06-21T15:18:56Zoai:repositorio.unesp.br:11449/173769Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T18:15:51.779428Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
title |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
spellingShingle |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction Marcos, Caroline Maria [UNESP] EF-Tu fibronectin fungal-host interaction moonlighting protein Paracoccidioides brasiliensis plasminogen |
title_short |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
title_full |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
title_fullStr |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
title_full_unstemmed |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
title_sort |
Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction |
author |
Marcos, Caroline Maria [UNESP] |
author_facet |
Marcos, Caroline Maria [UNESP] de Oliveira, Haroldo Cesar [UNESP] da Silva, Julhiany de Fátima [UNESP] Assato, Patricia Akemi [UNESP] Yamazaki, Daniella Sayuri [UNESP] da Silva, Rosângela Aparecida Moraes [UNESP] Santos, Cláudia Tavares [UNESP] Santos-Filho, Norival Alves [UNESP] Portuondo, Deivys Leandro [UNESP] Mendes-Giannini, Maria José Soares [UNESP] Fusco-Almeida, Ana Marisa [UNESP] |
author_role |
author |
author2 |
de Oliveira, Haroldo Cesar [UNESP] da Silva, Julhiany de Fátima [UNESP] Assato, Patricia Akemi [UNESP] Yamazaki, Daniella Sayuri [UNESP] da Silva, Rosângela Aparecida Moraes [UNESP] Santos, Cláudia Tavares [UNESP] Santos-Filho, Norival Alves [UNESP] Portuondo, Deivys Leandro [UNESP] Mendes-Giannini, Maria José Soares [UNESP] Fusco-Almeida, Ana Marisa [UNESP] |
author2_role |
author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Marcos, Caroline Maria [UNESP] de Oliveira, Haroldo Cesar [UNESP] da Silva, Julhiany de Fátima [UNESP] Assato, Patricia Akemi [UNESP] Yamazaki, Daniella Sayuri [UNESP] da Silva, Rosângela Aparecida Moraes [UNESP] Santos, Cláudia Tavares [UNESP] Santos-Filho, Norival Alves [UNESP] Portuondo, Deivys Leandro [UNESP] Mendes-Giannini, Maria José Soares [UNESP] Fusco-Almeida, Ana Marisa [UNESP] |
dc.subject.por.fl_str_mv |
EF-Tu fibronectin fungal-host interaction moonlighting protein Paracoccidioides brasiliensis plasminogen |
topic |
EF-Tu fibronectin fungal-host interaction moonlighting protein Paracoccidioides brasiliensis plasminogen |
description |
Paracoccidioides spp., which are temperature-dependent dimorphic fungi, are responsible for the most prevalent human systemic mycosis in Latin America, the paracoccidioidomycosis. The aim of this study was to characterise the involvement of elongation factor Tu (EF-Tu) in Paracoccidioides brasiliensis-host interaction. Adhesive properties were examined using recombinant PbEF-Tu proteins and the respective polyclonal anti-rPbEF-Tu antibody. Immunogold analysis demonstrated the surface location of EF-Tu in P. brasiliensis. Moreover, PbEF-Tu was found to bind to fibronectin and plasminogen by enzyme-linked immunosorbent assay, and it was determined that the binding to plasminogen is at least partly dependent on lysine residues and ionic interactions. To verify the participation of EF-Tu in the interaction of P. brasiliensis with pneumocytes, we blocked the respective protein with an anti-rPbEF-Tu antibody and evaluated the consequences on the interaction index by flow cytometry. During the interaction, we observed a decrease of 2- and 3-fold at 8 and 24 h, respectively, suggesting the contribution of EF-Tu in fungal adhesion/invasion. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-11-01 2018-12-11T17:07:39Z 2018-12-11T17:07:39Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1093/femsyr/fow079 FEMS Yeast Research, v. 16, n. 7, 2016. 1567-1364 1567-1356 http://hdl.handle.net/11449/173769 10.1093/femsyr/fow079 2-s2.0-84994717965 2-s2.0-84994717965.pdf |
url |
http://dx.doi.org/10.1093/femsyr/fow079 http://hdl.handle.net/11449/173769 |
identifier_str_mv |
FEMS Yeast Research, v. 16, n. 7, 2016. 1567-1364 1567-1356 10.1093/femsyr/fow079 2-s2.0-84994717965 2-s2.0-84994717965.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
FEMS Yeast Research 1,308 1,308 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1808128913340104704 |