Role of Leishmania (Leishmania) amazonensis amastigote glycosphingolipids in macrophage infectivity

Detalhes bibliográficos
Autor(a) principal: Tanaka, Améria Kaori [UNIFESP]
Data de Publicação: 2007
Outros Autores: Gorin, Philip Albert James, Takahashi, Helio Kiyoshi [UNIFESP], Straus, Anita Hilda [UNIFESP]
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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spelling 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
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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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dc.publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
publisher.none.fl_str_mv Associação Brasileira de Divulgação Científica
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