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

Abstract

The aim of the present study was to demonstrate the presence of a-Lfucosidase in Trypanosoma cruzi. Immunocytochemical and biochemical techniques were used to localize and characterize a membrane-associated, neutral-pH-optimum, a-L-fucosidase from the parasite. Light and electron microscopy localized the a-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 a-L-fucosidase activity was associated with epimastigote membrane solubilized with 1 M NaCl or 1% Triton X-100, suggesting that a-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-a-Lfucoside 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-a-L-fucosidase antibodies.

Human collectin-11 (COLEC11) and its synergic genetic interaction with MASP2 are associated with the pathophysiology of Chagas Disease

“…The infective T . cruzi metacyclic trypomastigote has a broad range of carbohydrates on its surface, including mannose, N-acetyl-D-glucosamine, galactose, and fucose on glycosylated proteins [16]. These glycoconjugates act as pathogen-associated molecular patterns (PAMPs), allowing the PRMs to interact with them [9,17].…”