Immunocytochemical and biochemical detection of alpha-L-fucosidase in Trypanosoma cruzi
Braz. j. med. biol. res
; 36(5): 595-603, May 2003. ilus, tab, graf
Article
em En
| LILACS
| ID: lil-331462
Biblioteca responsável:
BR1.1
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 percent of the alpha-L-fucosidase activity was associated with epimastigote membrane solubilized with 1 M NaCl or 1 percent 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
Texto completo:
1
Coleções:
01-internacional
Base de dados:
LILACS
Assunto principal:
Trypanosoma cruzi
/
Alfa-L-Fucosidase
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
Braz. j. med. biol. res
Assunto da revista:
BIOLOGIA
/
MEDICINA
Ano de publicação:
2003
Tipo de documento:
Article
/
Project document
País de afiliação:
Brasil
País de publicação:
Brasil