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(Referência obtida automaticamente do SciELO, por meio da informação sobre o financiamento pela FAPESP e o número do processo correspondente, incluída na publicação pelos autores.)

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

Texto completo
Autor(es):
Tanaka, A. K. ; Gorin, P. A. J. ; Takahashi, H. K. [3] ; Straus, A. H.
Número total de Autores: 4
Tipo de documento: Artigo Científico
Fonte: Brazilian Journal of Medical and Biological Research; v. 40, n. 6, p. 799-806, June 2007.
Área do conhecimento: Ciências Biológicas - Bioquímica
Assunto(s):Glicoesfingolipídeos   Proteínas de transporte   Protozoa   Leishmania mexicana
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. (AU)

Processo FAPESP: 03/11274-2 - Glicoconjugados de tripanossomatídeos: papel na interação parasita-hospedeiro
Beneficiário:Anita Hilda Straus Takahashi
Linha de fomento: Auxílio à Pesquisa - Regular
Processo FAPESP: 06/07005-4 - Microdomínios de membrana ricos em (glico) (esfingo) lipídeos e esteróis: organização, função e sinalização
Beneficiário:Helio Kiyoshi Takahashi
Linha de fomento: Auxílio à Pesquisa - Temático
Processo FAPESP: 01/13755-2 - Papel dos glicoesfingolipídeos e proteofosfoglicanos na interação Leishmania (Leishmania) amazonensis-macrófago: caracterização de novos antígenos e receptores
Beneficiário:Améria Kaori Tanaka
Linha de fomento: Bolsas no Brasil - Doutorado