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

Detalhes bibliográficos
Autor(a) principal: Cisalpino, Patricia Silva [UNIFESP]
Data de Publicação: 1996
Outros Autores: Puccia, Rosana [UNIFESP], Yamauchi, Lucy Megumi [UNIFESP], Cano, Maria Isabel Nogueira [UNIFESP], Silveira, Jose Franco da [UNIFESP], Travassos, Luiz Rodolpho [UNIFESP]
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: http://doi.org/10.1074/jbc.271.8.4553
http://repositorio.unifesp.br/handle/11600/44207
Resumo: The 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.
id UFSP_691be0f37fd17cfc0126891b519a3605
oai_identifier_str oai:repositorio.unifesp.br/:11600/44207
network_acronym_str UFSP
network_name_str Repositório Institucional da UNIFESP
repository_id_str 3465
spelling Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensisThe 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.UNIV FED SAO PAULO,DEPT MICROBIOL IMUNOL & PARASITOL,DISCIPLINA BIOL CELULAR,BR-04023062 SAO PAULO,SP,BRAZILUNIV FED SAO PAULO,DEPT MICROBIOL IMUNOL & PARASITOL,DISCIPLINA BIOL CELULAR,BR-04023062 SAO PAULO,SP,BRAZILWeb of ScienceAmer Soc Biochemistry Molecular Biology IncUniversidade Federal de São Paulo (UNIFESP)Cisalpino, Patricia Silva [UNIFESP]Puccia, Rosana [UNIFESP]Yamauchi, Lucy Megumi [UNIFESP]Cano, Maria Isabel Nogueira [UNIFESP]Silveira, Jose Franco da [UNIFESP]Travassos, Luiz Rodolpho [UNIFESP]2018-06-15T17:53:05Z2018-06-15T17:53:05Z1996-02-23info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion4553-4560http://doi.org/10.1074/jbc.271.8.4553Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 8, p. 4553-4560, 1996.10.1074/jbc.271.8.45530021-9258http://repositorio.unifesp.br/handle/11600/44207WOS:A1996TW96000089engJournal Of Biological Chemistryinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-05-02T13:44:28Zoai:repositorio.unifesp.br/:11600/44207Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-05-02T13:44:28Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
title Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
spellingShingle Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
Cisalpino, Patricia Silva [UNIFESP]
title_short Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
title_full Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
title_fullStr Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
title_full_unstemmed Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
title_sort Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
author Cisalpino, Patricia Silva [UNIFESP]
author_facet Cisalpino, Patricia Silva [UNIFESP]
Puccia, Rosana [UNIFESP]
Yamauchi, Lucy Megumi [UNIFESP]
Cano, Maria Isabel Nogueira [UNIFESP]
Silveira, Jose Franco da [UNIFESP]
Travassos, Luiz Rodolpho [UNIFESP]
author_role author
author2 Puccia, Rosana [UNIFESP]
Yamauchi, Lucy Megumi [UNIFESP]
Cano, Maria Isabel Nogueira [UNIFESP]
Silveira, Jose Franco da [UNIFESP]
Travassos, Luiz Rodolpho [UNIFESP]
author2_role author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Cisalpino, Patricia Silva [UNIFESP]
Puccia, Rosana [UNIFESP]
Yamauchi, Lucy Megumi [UNIFESP]
Cano, Maria Isabel Nogueira [UNIFESP]
Silveira, Jose Franco da [UNIFESP]
Travassos, Luiz Rodolpho [UNIFESP]
description The 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.
publishDate 1996
dc.date.none.fl_str_mv 1996-02-23
2018-06-15T17:53:05Z
2018-06-15T17:53:05Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://doi.org/10.1074/jbc.271.8.4553
Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 8, p. 4553-4560, 1996.
10.1074/jbc.271.8.4553
0021-9258
http://repositorio.unifesp.br/handle/11600/44207
WOS:A1996TW96000089
url http://doi.org/10.1074/jbc.271.8.4553
http://repositorio.unifesp.br/handle/11600/44207
identifier_str_mv Journal Of Biological Chemistry. Bethesda: Amer Soc Biochemistry Molecular Biology Inc, v. 271, n. 8, p. 4553-4560, 1996.
10.1074/jbc.271.8.4553
0021-9258
WOS:A1996TW96000089
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Journal Of Biological Chemistry
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 4553-4560
dc.publisher.none.fl_str_mv Amer Soc Biochemistry Molecular Biology Inc
publisher.none.fl_str_mv Amer Soc Biochemistry Molecular Biology Inc
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
_version_ 1814268439752081408