Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity
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/11600/3777 http://dx.doi.org/10.1590/S0100-879X2006005000106 |
Resumo: | The role of glycosphingolipids (GSLs) present in amastigote forms of Leishmania (Leishmania) amazonensis during infection of macrophages was analyzed, with particular emphasis on GSLs presenting the terminal Galpß1-3Galpa disaccharide. Macrophage invasion by L. (L.) amazonensis amastigotes was reduced by 37% when the disaccharide Galpß1-3Galp (1 mM) was added to the culture medium. The putative macrophage receptor/lectin for ß-Gal-globotriaosylceramide (Galpß1-3Galpa1-4Galpß1-4Glc pß1-1Cer) and other structurally related GSLs from L. (L.) amazonensis amastigotes were analyzed by micelles and parasite binding assay to peritoneal macrophage proteins fractionated by SDS-PAGE under nonreducing conditions. Micelles containing purified amastigote GSLs or a suspention of L. (L.) amazonensis amastigotes fixed with 2% formaldehyde were incubated with nitrocellulose membrane containing the macrophage proteins transferred by Western blotting. Binding of micelles containing purified GSLs from amastigote forms or fixed L. (L.) amazonensis amastigotes to nitrocellulose membrane was probed using monoclonal antibody ST-3, which recognizes the glycoepitope Galpß1-3Galpa1-R present either in the micelle preparation or on the amastigote surface. Macrophage protein with molecular mass ~30 kDa bound the amastigote GSL and appeared to be a doublet on electrophoresis. The specificity of this interaction was confirmed using fixed L. (L.) chagasi amastigotes, which do not express GSLs such as ß-Galp-globotriaosylceramides, and which do not bind to 30-kDa protein. |
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Tanaka, Améria Kaori [UNIFESP]Gorin, Philip Albert JamesTakahashi, Helio Kiyoshi [UNIFESP]Straus, Anita Hilda [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Universidade Federal do Paraná Departamento de Bioquímica2015-06-14T13:36:59Z2015-06-14T13:36:59Z2007-06-01Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 40, n. 6, p. 799-806, 2007.0100-879Xhttp://repositorio.unifesp.br/handle/11600/3777http://dx.doi.org/10.1590/S0100-879X2006005000106S0100-879X2007000600008.pdfS0100-879X200700060000810.1590/S0100-879X2006005000106WOS:000248409900008The role of glycosphingolipids (GSLs) present in amastigote forms of Leishmania (Leishmania) amazonensis during infection of macrophages was analyzed, with particular emphasis on GSLs presenting the terminal Galpß1-3Galpa disaccharide. Macrophage invasion by L. (L.) amazonensis amastigotes was reduced by 37% when the disaccharide Galpß1-3Galp (1 mM) was added to the culture medium. The putative macrophage receptor/lectin for ß-Gal-globotriaosylceramide (Galpß1-3Galpa1-4Galpß1-4Glc pß1-1Cer) and other structurally related GSLs from L. (L.) amazonensis amastigotes were analyzed by micelles and parasite binding assay to peritoneal macrophage proteins fractionated by SDS-PAGE under nonreducing conditions. Micelles containing purified amastigote GSLs or a suspention of L. (L.) amazonensis amastigotes fixed with 2% formaldehyde were incubated with nitrocellulose membrane containing the macrophage proteins transferred by Western blotting. Binding of micelles containing purified GSLs from amastigote forms or fixed L. (L.) amazonensis amastigotes to nitrocellulose membrane was probed using monoclonal antibody ST-3, which recognizes the glycoepitope Galpß1-3Galpa1-R present either in the micelle preparation or on the amastigote surface. Macrophage protein with molecular mass ~30 kDa bound the amastigote GSL and appeared to be a doublet on electrophoresis. The specificity of this interaction was confirmed using fixed L. (L.) chagasi amastigotes, which do not express GSLs such as ß-Galp-globotriaosylceramides, and which do not bind to 30-kDa protein.Universidade Federal de São Paulo (UNIFESP) Escola Paulista de Medicina Departamento de BioquímicaUniversidade Federal do Paraná Departamento de BioquímicaUNIFESP, EPM, Depto. de BioquímicaSciELO799-806engAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological ResearchLeishmania (Leishmania) amazonensisAmastigoteGlycosphingolipidMacrophage binding proteinRole of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivityinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALS0100-879X2007000600008.pdfapplication/pdf607409${dspace.ui.url}/bitstream/11600/3777/1/S0100-879X2007000600008.pdf17e20a3769f4599fa22f20a2a9f7ef5bMD51open accessTEXTS0100-879X2007000600008.pdf.txtS0100-879X2007000600008.pdf.txtExtracted texttext/plain31061${dspace.ui.url}/bitstream/11600/3777/2/S0100-879X2007000600008.pdf.txt21c1d9ac9b221e416b97f38489cd5ba1MD52open access11600/37772021-09-29 14:52:14.591open accessoai:repositorio.unifesp.br:11600/3777Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:13:19.706045Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
title |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
spellingShingle |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity Tanaka, Améria Kaori [UNIFESP] Leishmania (Leishmania) amazonensis Amastigote Glycosphingolipid Macrophage binding protein |
title_short |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
title_full |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
title_fullStr |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
title_full_unstemmed |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
title_sort |
Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity |
author |
Tanaka, Améria Kaori [UNIFESP] |
author_facet |
Tanaka, Améria Kaori [UNIFESP] Gorin, Philip Albert James Takahashi, Helio Kiyoshi [UNIFESP] Straus, Anita Hilda [UNIFESP] |
author_role |
author |
author2 |
Gorin, Philip Albert James Takahashi, Helio Kiyoshi [UNIFESP] Straus, Anita Hilda [UNIFESP] |
author2_role |
author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Universidade Federal do Paraná Departamento de Bioquímica |
dc.contributor.author.fl_str_mv |
Tanaka, Améria Kaori [UNIFESP] Gorin, Philip Albert James Takahashi, Helio Kiyoshi [UNIFESP] Straus, Anita Hilda [UNIFESP] |
dc.subject.eng.fl_str_mv |
Leishmania (Leishmania) amazonensis Amastigote Glycosphingolipid Macrophage binding protein |
topic |
Leishmania (Leishmania) amazonensis Amastigote Glycosphingolipid Macrophage binding protein |
description |
The role of glycosphingolipids (GSLs) present in amastigote forms of Leishmania (Leishmania) amazonensis during infection of macrophages was analyzed, with particular emphasis on GSLs presenting the terminal Galpß1-3Galpa disaccharide. Macrophage invasion by L. (L.) amazonensis amastigotes was reduced by 37% when the disaccharide Galpß1-3Galp (1 mM) was added to the culture medium. The putative macrophage receptor/lectin for ß-Gal-globotriaosylceramide (Galpß1-3Galpa1-4Galpß1-4Glc pß1-1Cer) and other structurally related GSLs from L. (L.) amazonensis amastigotes were analyzed by micelles and parasite binding assay to peritoneal macrophage proteins fractionated by SDS-PAGE under nonreducing conditions. Micelles containing purified amastigote GSLs or a suspention of L. (L.) amazonensis amastigotes fixed with 2% formaldehyde were incubated with nitrocellulose membrane containing the macrophage proteins transferred by Western blotting. Binding of micelles containing purified GSLs from amastigote forms or fixed L. (L.) amazonensis amastigotes to nitrocellulose membrane was probed using monoclonal antibody ST-3, which recognizes the glycoepitope Galpß1-3Galpa1-R present either in the micelle preparation or on the amastigote surface. Macrophage protein with molecular mass ~30 kDa bound the amastigote GSL and appeared to be a doublet on electrophoresis. The specificity of this interaction was confirmed using fixed L. (L.) chagasi amastigotes, which do not express GSLs such as ß-Galp-globotriaosylceramides, and which do not bind to 30-kDa protein. |
publishDate |
2007 |
dc.date.issued.fl_str_mv |
2007-06-01 |
dc.date.accessioned.fl_str_mv |
2015-06-14T13:36:59Z |
dc.date.available.fl_str_mv |
2015-06-14T13:36:59Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 40, n. 6, p. 799-806, 2007. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/3777 http://dx.doi.org/10.1590/S0100-879X2006005000106 |
dc.identifier.issn.none.fl_str_mv |
0100-879X |
dc.identifier.file.none.fl_str_mv |
S0100-879X2007000600008.pdf |
dc.identifier.scielo.none.fl_str_mv |
S0100-879X2007000600008 |
dc.identifier.doi.none.fl_str_mv |
10.1590/S0100-879X2006005000106 |
dc.identifier.wos.none.fl_str_mv |
WOS:000248409900008 |
identifier_str_mv |
Brazilian Journal of Medical and Biological Research. Associação Brasileira de Divulgação Científica, v. 40, n. 6, p. 799-806, 2007. 0100-879X S0100-879X2007000600008.pdf S0100-879X2007000600008 10.1590/S0100-879X2006005000106 WOS:000248409900008 |
url |
http://repositorio.unifesp.br/handle/11600/3777 http://dx.doi.org/10.1590/S0100-879X2006005000106 |
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eng |
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Brazilian Journal of Medical and Biological Research |
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openAccess |
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799-806 |
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Associação Brasileira de Divulgação Científica |
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Associação Brasileira de Divulgação Científica |
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