TY - JOUR
T1 - Echinococcus granulosus: Antigen characterization by chemical treatment and enzymatic deglycosylation
AU - March, F.
AU - Enrich, C.
AU - Mercader, M.
AU - Sánchez, F.
AU - Muñoz, C.
AU - Coll, P.
AU - Prats, G.
PY - 1991/1/1
Y1 - 1991/1/1
N2 - Parasite antigenic fractions obtained by biochemical purification of sheep hydatid fluid were subjected to enzymatic digestion. The relative mobilities of the 5 and B antigens, before and after treatment, were analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Antigenic fractions transferred to nitrocellulose were also treated with sodium metaperiodate and concanavalin A. The results indicate that antigen 5 contains a substantial amount of carbohydrates covalently linked to a polypeptide backbone, which strongly bind to concanavalin A and is removed by N-glycosidase F (PNGase F). Antigen 5 possesses complex N-linked oligosaccharides (PNGase F sensitive), without terminal N-acetyl-d-glucosamine residues (N-acetyl-d-glucosaminidase nonsensitive) and has no high-mannose oligosaccharides (endo-β-N-acetylglucosaminidase H nonsensitive). In contrast, the antigen B of low molecular weight is not susceptible to either enzymatic digestions (PNGase F, Endo H, and N-acetyl-d-glucosaminidase) or sodium metaperiodate oxidation and it does not bind to concanavalin A. Polyclonal antibodies prepared against the two antigens reacted with the deglycosylated antigen 5 in Western blot. The dominant epitopes are, therefore, polypeptides, although the presence of carbohydrate epitopes in the native glycoproteins cannot be excluded. © 1991.
AB - Parasite antigenic fractions obtained by biochemical purification of sheep hydatid fluid were subjected to enzymatic digestion. The relative mobilities of the 5 and B antigens, before and after treatment, were analyzed by polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Antigenic fractions transferred to nitrocellulose were also treated with sodium metaperiodate and concanavalin A. The results indicate that antigen 5 contains a substantial amount of carbohydrates covalently linked to a polypeptide backbone, which strongly bind to concanavalin A and is removed by N-glycosidase F (PNGase F). Antigen 5 possesses complex N-linked oligosaccharides (PNGase F sensitive), without terminal N-acetyl-d-glucosamine residues (N-acetyl-d-glucosaminidase nonsensitive) and has no high-mannose oligosaccharides (endo-β-N-acetylglucosaminidase H nonsensitive). In contrast, the antigen B of low molecular weight is not susceptible to either enzymatic digestions (PNGase F, Endo H, and N-acetyl-d-glucosaminidase) or sodium metaperiodate oxidation and it does not bind to concanavalin A. Polyclonal antibodies prepared against the two antigens reacted with the deglycosylated antigen 5 in Western blot. The dominant epitopes are, therefore, polypeptides, although the presence of carbohydrate epitopes in the native glycoproteins cannot be excluded. © 1991.
KW - Echinococcus granulosus
U2 - 10.1016/0014-4894(91)90067-7
DO - 10.1016/0014-4894(91)90067-7
M3 - Article
VL - 73
SP - 433
EP - 439
JO - Experimental Parasitology
JF - Experimental Parasitology
SN - 0014-4894
IS - 4
ER -