Trypanosoma cruzi glycoprotein 72: immunological analysis and cellular localization.
Two monoclonal antibodies were used to biochemically characterize glycoprotein 72 (GP72) from Trypanosoma cruzi and to localize the protein in live and fixed parasites by indirect immunofluorescence and in thin section of parasites by immunogold electron microscopy. GP72 was shown in immunoblots to be specific for the epimastigote stage; the protein could not be detected in trypomastigotes. Each antibody reacted with a different epitope on the glycoprotein and deglycosylation of GP72 ablated reactivity with one of the antibodies. Indirect immunofluorescence and electron microscopic evaluation of parasite associated gold particles showed the presence of GP72 in the cell surface membrane including the flagellar pocket and the cytostome. In addition, cytoplasmic membrane vesicles of the endosomal-lysosomal system stained intensely.
Harth
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Department of Immunology and Infectious Diseases, Palo Alto Medical Foundation, California 94301.
Mills
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AA
Souto-Padrón
T
T
de Souza
W
W
D000818:Animals; D000911:Antibodies, Monoclonal; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D007124:Immunoenzyme Techniques; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase; D016253:Microscopy, Immunoelectron; D014349:Trypanosoma cruzi / Q000737:chemistry* / Q000254:growth & development; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000032:analysis / Q000276:immunology*
D000911:Antibodies, Monoclonal; D014643:Variant Surface Glycoproteins, Trypanosoma; D017038:Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase