Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes
Autor(a) principal: | |
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Data de Publicação: | 2007 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/29833 http://dx.doi.org/10.1128/IAI.00262-07 |
Resumo: | A member of the Trypanosoma cruzi gp82 family, expressed on metacyclic trypomastigote surface and identified by monoclonal antibody (MAb) 3F6, plays a key role in host cell invasion. Apart from the gp82 defined by MAb 3F6, no information is available on members of this protein family. From cDNA clones encoding gp82 proteins sharing 59.1% sequence identity, we produced the recombinant proteins J18 and C03, the former containing and the latter lacking the epitope for MAb 3F6. Polyclonal antibodies to J18 and C03 proteins were generated and used, along with MAb 3F6, to analyze the expression and cellular localization of gp82 family members in metacyclic forms of CL and G strains, which belong to highly divergent genetic groups. By two-dimensional gel electrophoresis and immunoblotting, molecules of 82 to 86 kDa, focusing at pH 4.6 to 5.4, and molecules of 72 to 88 kDa, focusing at pH 4.9 to 5.7, were visualized in CL and G strains, respectively. Flow cytometry and microscopic analysis revealed in both strains similar expression of MAb 3F6-reactive gp82 in live and permeabilized parasites, indicating its surface localization. the reaction of live parasites of both strains with anti-J18 antibodies was weaker than with MAb 3F6 and was increased by permeabilization. Anti-C03 antibodies bound predominantly to flagellar components in permeabilized G strain parasites, but in the CL strain the flagellum was not the preferential target for these antibodies. Host cell invasion of metacyclic forms was inhibited by J18 protein, as well as by MAb 3F6 and anti-J18 antibodies, but not by C03 protein or anti-C03 antibodies. |
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Atayde, Vanessa [UNIFESP]Cortez, Mauro [UNIFESP]Souza, Renata [UNIFESP]Silveira, Jose Franco da [UNIFESP]Yoshida, Nobuko [UNIFESP]Universidade Federal de São Paulo (UNIFESP)2016-01-24T13:48:49Z2016-01-24T13:48:49Z2007-07-01Infection and Immunity. Washington: Amer Soc Microbiology, v. 75, n. 7, p. 3264-3270, 2007.0019-9567http://repositorio.unifesp.br/handle/11600/29833http://dx.doi.org/10.1128/IAI.00262-07WOS000247707600006.pdf10.1128/IAI.00262-07WOS:000247707600006A member of the Trypanosoma cruzi gp82 family, expressed on metacyclic trypomastigote surface and identified by monoclonal antibody (MAb) 3F6, plays a key role in host cell invasion. Apart from the gp82 defined by MAb 3F6, no information is available on members of this protein family. From cDNA clones encoding gp82 proteins sharing 59.1% sequence identity, we produced the recombinant proteins J18 and C03, the former containing and the latter lacking the epitope for MAb 3F6. Polyclonal antibodies to J18 and C03 proteins were generated and used, along with MAb 3F6, to analyze the expression and cellular localization of gp82 family members in metacyclic forms of CL and G strains, which belong to highly divergent genetic groups. By two-dimensional gel electrophoresis and immunoblotting, molecules of 82 to 86 kDa, focusing at pH 4.6 to 5.4, and molecules of 72 to 88 kDa, focusing at pH 4.9 to 5.7, were visualized in CL and G strains, respectively. Flow cytometry and microscopic analysis revealed in both strains similar expression of MAb 3F6-reactive gp82 in live and permeabilized parasites, indicating its surface localization. the reaction of live parasites of both strains with anti-J18 antibodies was weaker than with MAb 3F6 and was increased by permeabilization. Anti-C03 antibodies bound predominantly to flagellar components in permeabilized G strain parasites, but in the CL strain the flagellum was not the preferential target for these antibodies. Host cell invasion of metacyclic forms was inhibited by J18 protein, as well as by MAb 3F6 and anti-J18 antibodies, but not by C03 protein or anti-C03 antibodies.Universidade Federal de São Paulo, Dept Microbiol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol, BR-04023062 São Paulo, BrazilWeb of Science3264-3270engAmer Soc MicrobiologyInfection and ImmunityExpression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000247707600006.pdfapplication/pdf371599${dspace.ui.url}/bitstream/11600/29833/1/WOS000247707600006.pdfe8858af4bf1b8dbcf59b13cd78d0c21cMD51open accessTEXTWOS000247707600006.pdf.txtWOS000247707600006.pdf.txtExtracted texttext/plain40407${dspace.ui.url}/bitstream/11600/29833/2/WOS000247707600006.pdf.txt3594f30d191e99124d975ee22a3531b9MD52open access11600/298332023-01-12 21:39:41.733open accessoai:repositorio.unifesp.br:11600/29833Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-01-13T00:39:41Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
title |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
spellingShingle |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes Atayde, Vanessa [UNIFESP] |
title_short |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
title_full |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
title_fullStr |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
title_full_unstemmed |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
title_sort |
Expression and cellular localization of molecules of the gp82 family in Trypanosoma cruzi metacyclic trypomastigotes |
author |
Atayde, Vanessa [UNIFESP] |
author_facet |
Atayde, Vanessa [UNIFESP] Cortez, Mauro [UNIFESP] Souza, Renata [UNIFESP] Silveira, Jose Franco da [UNIFESP] Yoshida, Nobuko [UNIFESP] |
author_role |
author |
author2 |
Cortez, Mauro [UNIFESP] Souza, Renata [UNIFESP] Silveira, Jose Franco da [UNIFESP] Yoshida, Nobuko [UNIFESP] |
author2_role |
author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) |
dc.contributor.author.fl_str_mv |
Atayde, Vanessa [UNIFESP] Cortez, Mauro [UNIFESP] Souza, Renata [UNIFESP] Silveira, Jose Franco da [UNIFESP] Yoshida, Nobuko [UNIFESP] |
description |
A member of the Trypanosoma cruzi gp82 family, expressed on metacyclic trypomastigote surface and identified by monoclonal antibody (MAb) 3F6, plays a key role in host cell invasion. Apart from the gp82 defined by MAb 3F6, no information is available on members of this protein family. From cDNA clones encoding gp82 proteins sharing 59.1% sequence identity, we produced the recombinant proteins J18 and C03, the former containing and the latter lacking the epitope for MAb 3F6. Polyclonal antibodies to J18 and C03 proteins were generated and used, along with MAb 3F6, to analyze the expression and cellular localization of gp82 family members in metacyclic forms of CL and G strains, which belong to highly divergent genetic groups. By two-dimensional gel electrophoresis and immunoblotting, molecules of 82 to 86 kDa, focusing at pH 4.6 to 5.4, and molecules of 72 to 88 kDa, focusing at pH 4.9 to 5.7, were visualized in CL and G strains, respectively. Flow cytometry and microscopic analysis revealed in both strains similar expression of MAb 3F6-reactive gp82 in live and permeabilized parasites, indicating its surface localization. the reaction of live parasites of both strains with anti-J18 antibodies was weaker than with MAb 3F6 and was increased by permeabilization. Anti-C03 antibodies bound predominantly to flagellar components in permeabilized G strain parasites, but in the CL strain the flagellum was not the preferential target for these antibodies. Host cell invasion of metacyclic forms was inhibited by J18 protein, as well as by MAb 3F6 and anti-J18 antibodies, but not by C03 protein or anti-C03 antibodies. |
publishDate |
2007 |
dc.date.issued.fl_str_mv |
2007-07-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T13:48:49Z |
dc.date.available.fl_str_mv |
2016-01-24T13:48:49Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Infection and Immunity. Washington: Amer Soc Microbiology, v. 75, n. 7, p. 3264-3270, 2007. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/29833 http://dx.doi.org/10.1128/IAI.00262-07 |
dc.identifier.issn.none.fl_str_mv |
0019-9567 |
dc.identifier.file.none.fl_str_mv |
WOS000247707600006.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1128/IAI.00262-07 |
dc.identifier.wos.none.fl_str_mv |
WOS:000247707600006 |
identifier_str_mv |
Infection and Immunity. Washington: Amer Soc Microbiology, v. 75, n. 7, p. 3264-3270, 2007. 0019-9567 WOS000247707600006.pdf 10.1128/IAI.00262-07 WOS:000247707600006 |
url |
http://repositorio.unifesp.br/handle/11600/29833 http://dx.doi.org/10.1128/IAI.00262-07 |
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eng |
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Amer Soc Microbiology |
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Amer Soc Microbiology |
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