Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi
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...
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| Format: | Article |
| Language: | English |
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Associação Brasileira de Divulgação Científica
2003-05-01
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| Series: | Brazilian Journal of Medical and Biological Research |
| Subjects: | |
| Online Access: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006 |
| _version_ | 1811207388261253120 |
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| author | L.C. Miletti M. Almeida-de-Faria W. Colli M.J.M. Alves |
| author_facet | L.C. Miletti M. Almeida-de-Faria W. Colli M.J.M. Alves |
| author_sort | L.C. Miletti |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-12T04:02:19Z |
| format | Article |
| id | doaj.art-e8323d8964ce40808db544a3e818235b |
| institution | Directory Open Access Journal |
| issn | 0100-879X 1414-431X |
| language | English |
| last_indexed | 2024-04-12T04:02:19Z |
| publishDate | 2003-05-01 |
| publisher | Associação Brasileira de Divulgação Científica |
| record_format | Article |
| series | Brazilian Journal of Medical and Biological Research |
| spelling | doaj.art-e8323d8964ce40808db544a3e818235b2022-12-22T03:48:41ZengAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research0100-879X1414-431X2003-05-0136559560310.1590/S0100-879X2003000500006Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruziL.C. MilettiM. Almeida-de-FariaW. ColliM.J.M. AlvesThe 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.http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006Trypanosoma cruziTrypanosomatidsFucosidaseFucoseGlycoproteinsGlycolipids |
| spellingShingle | L.C. Miletti M. Almeida-de-Faria W. Colli M.J.M. Alves Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi Brazilian Journal of Medical and Biological Research Trypanosoma cruzi Trypanosomatids Fucosidase Fucose Glycoproteins Glycolipids |
| title | 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_short | Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi |
| title_sort | immunocytochemical and biochemical detection of alpha l fucosidase in trypanosoma cruzi |
| topic | Trypanosoma cruzi Trypanosomatids Fucosidase Fucose Glycoproteins Glycolipids |
| url | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2003000500006 |
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