title: Identification and characterisation of elongation factor Tu, a novel protein involved in Paracoccidioides brasiliensis-host interaction.
abstract: 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.
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Caroline Maria
authors.initials: CM
authors.lastname: Marcos
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Haroldo Cesar
authors.initials: HC
authors.lastname: de Oliveira
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Julhiany de Fátima
authors.initials: JF
authors.lastname: da Silva
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Patricia Akemi
authors.initials: PA
authors.lastname: Assato
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Daniella Sayuri
authors.initials: DS
authors.lastname: Yamazaki
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Rosângela Aparecida Moraes
authors.initials: RA
authors.lastname: da Silva
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Cláudia Tavares
authors.initials: CT
authors.lastname: Santos
authors.affiliation: Instituto de Química, UNESP - Univ Estadual Paulista, Departamento de Bioquímica e Tecnologia Química, Unidade de Síntese, Estrutura e Caracterização de Peptídeos e Proteínas, Araraquara, São Paulo, Brasil.
authors.forename: Norival Alves
authors.initials: NA
authors.lastname: Santos-Filho
authors.affiliation: Faculdade de Ciências Farmacéuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Imunologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Deivys Leandro
authors.initials: DL
authors.lastname: Portuondo
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil.
authors.forename: Maria José Soares
authors.initials: MJ
authors.lastname: Mendes-Giannini
authors.affiliation: Faculdade de Ciências Farmacêuticas, UNESP - Univ Estadual Paulista, Departamento de Análises Clínicas, Laboratório de Micologia Clínica, Araraquara, São Paulo, Brasil ana.marisa@uol.com.br.
authors.forename: Ana Marisa
authors.initials: AM
authors.lastname: Fusco-Almeida
mesh_terms: D056809:Alveolar Epithelial Cells / Q000382:microbiology; D002448:Cell Adhesion; D002460:Cell Line; D005353:Fibronectins / Q000378:metabolism; D054884:Host-Pathogen Interactions; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism; D010228:Paracoccidioides / Q000201:enzymology* / Q000502:physiology; D010444:Peptide Elongation Factor Tu / Q000378:metabolism*; D010958:Plasminogen / Q000378:metabolism; D011485:Protein Binding; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D008565:Membrane Proteins; D037521:Virulence Factors; D010958:Plasminogen; D010444:Peptide Elongation Factor Tu
keywords: EF-Tu; Paracoccidioides brasiliensis; fibronectin; fungal–host interaction; moonlighting protein; plasminogen