Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi

Detalhes bibliográficos
Autor(a) principal: Miletti,L.C.
Data de Publicação: 2003
Outros Autores: Almeida-de-Faria,M., Colli,W., Alves,M.J.M.
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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spelling 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
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