Trypanosoma rangeli sialidase: kinetics of release and antigenic characterization.
The epimastigote stage of Trypanosoma rangeli release a sialidase with a high sialic acid hydrolysis capacity. We demonstrate that sialidase secretion is an active process that is reduced at low temperatures and in the presence of sodium azide. The enzyme is continuously released until certain maximally active concentrations are attained in the BHI culture medium when the parasite density reaches 2-3 x 10(6) cells. When introduced into culture medium already containing such enzyme levels, freshly harvested parasites do not secrete additional sialidase. These findings suggest a self-regulating mechanism and a biological role for the secreted T. rangeli sialidase. The secreted enzyme was purified to homogeneity by fractionation with ammonium sulphate and affinity chromatography. Antibodies raised against the purified molecule recognized antigens of similar molecular weights (73 kDa) in western immunoblotting analyses of T. rangeli and T. cruzi whole cell lysates. No antigenic recognition was recorded against T. cruzi active sialidase/trans-sialidase polypeptides or Clostridium perfringens and Vibrio cholerae commercial sialidases. These observations may indicate the expression of different antigenic domains in T. rangeli, T. cruzi and bacterial sialidases.
Center for Research and Diagnosis of Parasitic Diseases (CIDEP), Faculty of Medicine, University of Panama.
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A
Saldaña
O E
OE
Sousa
A
A
Orn
R A
RA
Harris
D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology*; D015153:Blotting, Western; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D009439:Neuraminidase / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism*; D019810:Sodium Azide / Q000494:pharmacology; D013696:Temperature; D014345:Trypanosoma / Q000201:enzymology* / Q000276:immunology
D000953:Antigens, Protozoan; D003470:Culture Media; D019810:Sodium Azide; D009439:Neuraminidase