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Trypanosoma cruzi glycoprotein 72: Immunological analysis and cellular localization
Authors:Guenter Harth  Alea A Mills  Thais Souto-Padrón  Wanderley de Souza
Institution:(1) Department of Immunology and Infectious Diseases, Research Institute, Palo Alto Medical Foundation, 94301 Palo Alto, California;(2) Department of Medicine, Stanford University School of Medicine, 94305 Stanford, California;(3) Laboratório de Ultraestrutura Celular e Microscopia Eletrônica, Instituto de Biofisica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Ilha do Fundão, 21941 Rio de Janeiro, Brazil
Abstract: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.
Keywords:T  cruzi  glycoprotein 72 (GP72)  stage specificity  electron microscopy
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