Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi
Autor(a) principal: | |
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Data de Publicação: | 2003 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Medical and Biological Research |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006 |
Resumo: | The aim of the present study was to demonstrate the presence of alpha-L-fucosidase in Trypanosoma cruzi. Immunocytochemical and biochemical techniques were used to localize and characterize a membrane-associated, neutral-pH-optimum, alpha-L-fucosidase from the parasite. Light and electron microscopy localized the alpha-L-fucosidase specifically on the surface of the parasite and on membranes in the posterior region of the epimastigote stage. Although much less intense, labeling was also detected on the surface of trypomastigotes. At least 50% of the alpha-L-fucosidase activity was associated with epimastigote membrane solubilized with 1 M NaCl or 1% Triton X-100, suggesting that alpha-L-fucosidase is peripherally associated with membranes. The enzyme from epimastigotes had a neutral pH optimum (near 7) but displayed low specific activity when p-nitrophenyl-alpha-L-fucoside was employed as substrate (0.028 U/mg protein for epimastigotes and 0.015 U/mg protein for tissue culture-derived trypomastigotes). Polyacrylamide gel electrophoresis and Western blotting analysis both showed an expected 50-kDa polypeptide which was immunoreactive with anti-alpha-L-fucosidase antibodies. |
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Brazilian Journal of Medical and Biological Research |
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Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruziTrypanosoma cruziTrypanosomatidsFucosidaseFucoseGlycoproteinsGlycolipidsThe aim of the present study was to demonstrate the presence of alpha-L-fucosidase in Trypanosoma cruzi. Immunocytochemical and biochemical techniques were used to localize and characterize a membrane-associated, neutral-pH-optimum, alpha-L-fucosidase from the parasite. Light and electron microscopy localized the alpha-L-fucosidase specifically on the surface of the parasite and on membranes in the posterior region of the epimastigote stage. Although much less intense, labeling was also detected on the surface of trypomastigotes. At least 50% of the alpha-L-fucosidase activity was associated with epimastigote membrane solubilized with 1 M NaCl or 1% Triton X-100, suggesting that alpha-L-fucosidase is peripherally associated with membranes. The enzyme from epimastigotes had a neutral pH optimum (near 7) but displayed low specific activity when p-nitrophenyl-alpha-L-fucoside was employed as substrate (0.028 U/mg protein for epimastigotes and 0.015 U/mg protein for tissue culture-derived trypomastigotes). Polyacrylamide gel electrophoresis and Western blotting analysis both showed an expected 50-kDa polypeptide which was immunoreactive with anti-alpha-L-fucosidase antibodies.Associação Brasileira de Divulgação Científica2003-05-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006Brazilian Journal of Medical and Biological Research v.36 n.5 2003reponame:Brazilian Journal of Medical and Biological Researchinstname:Associação Brasileira de Divulgação Científica (ABDC)instacron:ABDC10.1590/S0100-879X2003000500006info:eu-repo/semantics/openAccessMiletti,L.C.Almeida-de-Faria,M.Colli,W.Alves,M.J.M.eng2003-04-22T00:00:00Zoai:scielo:S0100-879X2003000500006Revistahttps://www.bjournal.org/https://old.scielo.br/oai/scielo-oai.phpbjournal@terra.com.br||bjournal@terra.com.br1414-431X0100-879Xopendoar:2003-04-22T00:00Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC)false |
dc.title.none.fl_str_mv |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
title |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
spellingShingle |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi Miletti,L.C. Trypanosoma cruzi Trypanosomatids Fucosidase Fucose Glycoproteins Glycolipids |
title_short |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
title_full |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
title_fullStr |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
title_full_unstemmed |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
title_sort |
Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
author |
Miletti,L.C. |
author_facet |
Miletti,L.C. Almeida-de-Faria,M. Colli,W. Alves,M.J.M. |
author_role |
author |
author2 |
Almeida-de-Faria,M. Colli,W. Alves,M.J.M. |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Miletti,L.C. Almeida-de-Faria,M. Colli,W. Alves,M.J.M. |
dc.subject.por.fl_str_mv |
Trypanosoma cruzi Trypanosomatids Fucosidase Fucose Glycoproteins Glycolipids |
topic |
Trypanosoma cruzi Trypanosomatids Fucosidase Fucose Glycoproteins Glycolipids |
description |
The aim of the present study was to demonstrate the presence of alpha-L-fucosidase in Trypanosoma cruzi. Immunocytochemical and biochemical techniques were used to localize and characterize a membrane-associated, neutral-pH-optimum, alpha-L-fucosidase from the parasite. Light and electron microscopy localized the alpha-L-fucosidase specifically on the surface of the parasite and on membranes in the posterior region of the epimastigote stage. Although much less intense, labeling was also detected on the surface of trypomastigotes. At least 50% of the alpha-L-fucosidase activity was associated with epimastigote membrane solubilized with 1 M NaCl or 1% Triton X-100, suggesting that alpha-L-fucosidase is peripherally associated with membranes. The enzyme from epimastigotes had a neutral pH optimum (near 7) but displayed low specific activity when p-nitrophenyl-alpha-L-fucoside was employed as substrate (0.028 U/mg protein for epimastigotes and 0.015 U/mg protein for tissue culture-derived trypomastigotes). Polyacrylamide gel electrophoresis and Western blotting analysis both showed an expected 50-kDa polypeptide which was immunoreactive with anti-alpha-L-fucosidase antibodies. |
publishDate |
2003 |
dc.date.none.fl_str_mv |
2003-05-01 |
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://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0100-879X2003000500006 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica |
publisher.none.fl_str_mv |
Associação Brasileira de Divulgação Científica |
dc.source.none.fl_str_mv |
Brazilian Journal of Medical and Biological Research v.36 n.5 2003 reponame:Brazilian Journal of Medical and Biological Research instname:Associação Brasileira de Divulgação Científica (ABDC) instacron:ABDC |
instname_str |
Associação Brasileira de Divulgação Científica (ABDC) |
instacron_str |
ABDC |
institution |
ABDC |
reponame_str |
Brazilian Journal of Medical and Biological Research |
collection |
Brazilian Journal of Medical and Biological Research |
repository.name.fl_str_mv |
Brazilian Journal of Medical and Biological Research - Associação Brasileira de Divulgação Científica (ABDC) |
repository.mail.fl_str_mv |
bjournal@terra.com.br||bjournal@terra.com.br |
_version_ |
1754302932157726720 |