Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis

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dc.contributor.authorCisalpino, Patricia Silva [UNIFESP]
dc.contributor.authorPuccia, Rosana [UNIFESP]
dc.contributor.authorYamauchi, Lucy Megumi [UNIFESP]
dc.contributor.authorCano, Maria Isabel Nogueira [UNIFESP]
dc.contributor.authorSilveira, Jose Franco da [UNIFESP]
dc.contributor.authorTravassos, Luiz Rodolpho [UNIFESP]
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2018-06-15T17:53:05Z
dc.date.available2018-06-15T17:53:05Z
dc.date.issued1996-02-23
dc.description.abstractThe 43,000-Da glycoprotein (gp43) of Paracoccidioides brasiliensis is an immunodominant antigen for antibody-dependent and immune cellular responses in patients with paracoccidioidomycosis. In order to identify the peptide epitopes involved in the immunological reactivities of the gp43 and to obtain highly specific recombinant molecules for diagnosis of the infection, genomic and cDNA clones representing the entire coding region of the antigen were sequenced. The gp43 open reading frame was found in a 1,329-base pair fragment with 2 exons interrupted by an intron of 78 nucleotides. The gene is present in very few copies per genome, as indicated by Southern blotting and chromosomal mega-restriction analysis. A single transcript of 1.5 kilobase pairs was verified in the yeast phase. The gene encodes a polypeptide of 416 amino acids (M(r) 45,947) with a leader peptide of 35 residues; the mature protein has at single N-glycosylation site. The deduced amino acid sequence showed similarities of 56-58% with exo-1,3-beta-D-glucanases hom Saccharomyces cerevisiae and Candida albicans. However, the gp43 is devoid of hydrolase activity and does not cross-react immunologically with the fungal glucanases. Internal and COOH-terminal gene fragments of the gp43 were expressed as recombinant fusion proteins, which reacted with antibodies elicited against the native antigen.en
dc.description.affiliationUNIV FED SAO PAULO,DEPT MICROBIOL IMUNOL & PARASITOL,DISCIPLINA BIOL CELULAR,BR-04023062 SAO PAULO,SP,BRAZIL
dc.description.affiliationUnifespUNIV FED SAO PAULO,DEPT MICROBIOL IMUNOL & PARASITOL,DISCIPLINA BIOL CELULAR,BR-04023062 SAO PAULO,SP,BRAZIL
dc.description.sourceWeb of Science
dc.format.extent4553-4560
dc.identifierhttp://doi.org/10.1074/jbc.271.8.4553
dc.identifier.citationJournal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 8, p. 4553-4560, 1996.
dc.identifier.doi10.1074/jbc.271.8.4553
dc.identifier.issn0021-9258
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/44207
dc.identifier.wosWOS:A1996TW96000089
dc.language.isoeng
dc.publisherAmer Soc Biochemistry Molecular Biology Inc
dc.relation.ispartofJournal Of Biological Chemistry
dc.rightsinfo:eu-repo/semantics/openAccess
dc.titleCloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensisen
dc.typeinfo:eu-repo/semantics/article
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