title: Structural and functional characterization of the recombinant thioredoxin reductase from Candida albicans as a potential target for vaccine and drug design.
abstract: The thioredoxin system plays a critical role in maintaining the cytoplasm redox state, participating in functions that are important to the cellular viability of fungi. Although functional and structural information on targets in human pathogenic fungi has been scarcely described in the literature, such studies are essential for in silico drug design and biotechnological applications. Therefore, the aims of the present study were to produce recombinant proteins of the thioredoxin system from Candida albicans and evaluate their possible use as prophylactic or alternative therapies against the most important pathogenic fungus associated with nosocomial infections. We focused on biochemical and structural analyses of recombinant thioredoxin reductase from C. albicans with His-tag (CaTrxR-His) for further biotechnology applications. Heterologous CaTrxR-His was efficiently expressed in the soluble fraction of the Escherichia coli lysate. CaTrxR-His was obtained with a high level of purity and presented specific enzymatic activity. Conformational changes of the protein were observed at different pHs and temperatures, with higher thermal stability at pH 8.0. The CaTrxR-His vaccine was shown to effectively induce high levels of CaTrxR-specific immunoglobulin G antibodies in Balb/c mice and reduce the renal fungal burden of experimental disseminated candidiasis in mice. These data may greatly impact future development strategies for vaccine and drug designs against C. albicans infection.
authors.affiliation: Divisão de Micologia Médica, Departamento de Análises Clínicas e Biomedicina, Universidade Estadual de Maringá, Av. Colombo, 5790, bloco J90 sala 11, 87020-900, Maringá, Paraná, Brazil.
authors.forename: Janine Silva Ribeiro
authors.initials: JS
authors.lastname: Godoy
authors.affiliation: Divisão de Micologia Médica, Departamento de Análises Clínicas e Biomedicina, Universidade Estadual de Maringá, Av. Colombo, 5790, bloco J90 sala 11, 87020-900, Maringá, Paraná, Brazil. eskcotica@uem.br.
authors.forename: Érika Seki
authors.initials: ÉS
authors.lastname: Kioshima
authors.affiliation: Departamento de Biologia, Universidade do Estado do Mato Grosso - UNEMAT, Nova Xavantina, Mato Grosso, Brazil.
authors.forename: Ana Karina Rodrigues
authors.initials: AK
authors.lastname: Abadio
authors.affiliation: Laboratório de Biologia Molecular, Departamento de Biologia Celular, Universidade de Brasília, Brasília, Distrito Federal, Brazil.
authors.forename: Maria Sueli Soares
authors.initials: MS
authors.lastname: Felipe
authors.affiliation: Laboratório de Biofísica, Departamento de Biologia Celular, Universidade de Brasília, Brasília, Distrito Federal, Brazil.
authors.forename: Sonia Maria
authors.initials: SM
authors.lastname: de Freitas
authors.affiliation: Divisão de Micologia Médica, Departamento de Análises Clínicas e Biomedicina, Universidade Estadual de Maringá, Av. Colombo, 5790, bloco J90 sala 11, 87020-900, Maringá, Paraná, Brazil.
authors.forename: Terezinha Inez Estivalet
authors.initials: TI
authors.lastname: Svidzinski
mesh_terms: D000818:Animals; D000908:Antibodies, Fungal / Q000097:blood; D000946:Antigens, Fungal / Q000235:genetics / Q000378:metabolism; D002176:Candida albicans / Q000201:enzymology* / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000097:blood; D008807:Mice, Inbred BALB C; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013696:Temperature; D013880:Thioredoxin-Disulfide Reductase / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000908:Antibodies, Fungal; D000946:Antigens, Fungal; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D013880:Thioredoxin-Disulfide Reductase
keywords: Candida albicans; Recombinant protein; Thioredoxin reductase; Vaccine