Cloning, characterization, and epitope expression of the major diagnostic antigen of Paracoccidioides brasiliensis
Autor(a) principal: | |
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Data de Publicação: | 1996 |
Outros Autores: | , , , , |
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. |
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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 |