The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies

The Tc‐85 glycoprotein, specific for the infective stage of Trypanosoma cruzi, is anchored via glycosylphosphatidylinositol. The protein was purified from parasites, labeled metabolically with palmitic acid, by immunoprecipitation with the H 1 A 10 monoclonal antibody or by affinity column chromatog...

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Autores principales:	Couto, Alicia Susana, Muchnik de Lederkremer, Rosa María
Publicado:	1993
Materias:	glycoprotein glycosylphosphatidylinositol article immunoprecipitation nonhuman priority journal protein structure trypanosoma cruzi trypomastigote Animal Antibodies, Monoclonal Blotting, Western Carbohydrate Sequence Chromatography, Affinity Cross Reactions Electrophoresis, Polyacrylamide Gel Glycoproteins Glycosylphosphatidylinositols Lipids Molecular Sequence Data Palmitic Acid Palmitic Acids Precipitin Tests Protozoan Proteins Support, Non-U.S. Gov't Trypanosoma cruzi Wheat Germ Agglutinins Triticum aestivum Trypanosoma Trypanosoma brucei brucei
Acceso en línea:	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00142956_v217_n2_p597_COUTO http://hdl.handle.net/20.500.12110/paper_00142956_v217_n2_p597_COUTO
Aporte de:	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires

id	paper:paper_00142956_v217_n2_p597_COUTO
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spelling	paper:paper_00142956_v217_n2_p597_COUTO2023-06-08T14:36:42Z The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies Couto, Alicia Susana Muchnik de Lederkremer, Rosa María glycoprotein glycosylphosphatidylinositol article immunoprecipitation nonhuman priority journal protein structure trypanosoma cruzi trypomastigote Animal Antibodies, Monoclonal Blotting, Western Carbohydrate Sequence Chromatography, Affinity Cross Reactions Electrophoresis, Polyacrylamide Gel Glycoproteins Glycosylphosphatidylinositols Lipids Molecular Sequence Data Palmitic Acid Palmitic Acids Precipitin Tests Protozoan Proteins Support, Non-U.S. Gov't Trypanosoma cruzi Wheat Germ Agglutinins Triticum aestivum Trypanosoma Trypanosoma brucei brucei Trypanosoma cruzi The Tc‐85 glycoprotein, specific for the infective stage of Trypanosoma cruzi, is anchored via glycosylphosphatidylinositol. The protein was purified from parasites, labeled metabolically with palmitic acid, by immunoprecipitation with the H 1 A 10 monoclonal antibody or by affinity column chromatography on wheat germ agglutinin. Antisera to the soluble form of the variant surface glycoprotein of Trypanosoma brucei brucei cross‐reacted with Tc‐85 when the immunoprecipitate was analysed by Western blotting. The reaction was intensified upon previous incubation of the glycoprotein with phosphatidylinositol‐specific phospholipase C. Such recognition was abolished when the cyclic phosphate was opened by mild acid treatment. The lipid cleaved by phospholipase C digestion, was identified as 1‐O‐hexadecylglycerol by reverse‐phase thin‐layer chromatography. The glycan core was deaminated and chemically labeled by reduction with NaB 3 H 4 . The labeled glycoprotein was exhaustively treated with pronase and dephosphorylated with 50% HF. Although microheterogeneity of the oligosaccharide moiety was apparent, by thin layer chromatography, a main spot coincident with Man(α1–2) Man(α1–6) Man(α1–4) anhydromannitol was shown, consistent with the conserved core structure of all glycosylphosphatidylinositol anchors analysed to date. Copyright © 1993, Wiley Blackwell. All rights reserved Fil:COUTO, A.S. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:DE LEDERKREMER, R.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 1993 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00142956_v217_n2_p597_COUTO http://hdl.handle.net/20.500.12110/paper_00142956_v217_n2_p597_COUTO
institution	Universidad de Buenos Aires
institution_str	I-28
repository_str	R-134
collection	Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic	glycoprotein glycosylphosphatidylinositol article immunoprecipitation nonhuman priority journal protein structure trypanosoma cruzi trypomastigote Animal Antibodies, Monoclonal Blotting, Western Carbohydrate Sequence Chromatography, Affinity Cross Reactions Electrophoresis, Polyacrylamide Gel Glycoproteins Glycosylphosphatidylinositols Lipids Molecular Sequence Data Palmitic Acid Palmitic Acids Precipitin Tests Protozoan Proteins Support, Non-U.S. Gov't Trypanosoma cruzi Wheat Germ Agglutinins Triticum aestivum Trypanosoma Trypanosoma brucei brucei Trypanosoma cruzi
spellingShingle	glycoprotein glycosylphosphatidylinositol article immunoprecipitation nonhuman priority journal protein structure trypanosoma cruzi trypomastigote Animal Antibodies, Monoclonal Blotting, Western Carbohydrate Sequence Chromatography, Affinity Cross Reactions Electrophoresis, Polyacrylamide Gel Glycoproteins Glycosylphosphatidylinositols Lipids Molecular Sequence Data Palmitic Acid Palmitic Acids Precipitin Tests Protozoan Proteins Support, Non-U.S. Gov't Trypanosoma cruzi Wheat Germ Agglutinins Triticum aestivum Trypanosoma Trypanosoma brucei brucei Trypanosoma cruzi Couto, Alicia Susana Muchnik de Lederkremer, Rosa María The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies
topic_facet	glycoprotein glycosylphosphatidylinositol article immunoprecipitation nonhuman priority journal protein structure trypanosoma cruzi trypomastigote Animal Antibodies, Monoclonal Blotting, Western Carbohydrate Sequence Chromatography, Affinity Cross Reactions Electrophoresis, Polyacrylamide Gel Glycoproteins Glycosylphosphatidylinositols Lipids Molecular Sequence Data Palmitic Acid Palmitic Acids Precipitin Tests Protozoan Proteins Support, Non-U.S. Gov't Trypanosoma cruzi Wheat Germ Agglutinins Triticum aestivum Trypanosoma Trypanosoma brucei brucei Trypanosoma cruzi
description	The Tc‐85 glycoprotein, specific for the infective stage of Trypanosoma cruzi, is anchored via glycosylphosphatidylinositol. The protein was purified from parasites, labeled metabolically with palmitic acid, by immunoprecipitation with the H 1 A 10 monoclonal antibody or by affinity column chromatography on wheat germ agglutinin. Antisera to the soluble form of the variant surface glycoprotein of Trypanosoma brucei brucei cross‐reacted with Tc‐85 when the immunoprecipitate was analysed by Western blotting. The reaction was intensified upon previous incubation of the glycoprotein with phosphatidylinositol‐specific phospholipase C. Such recognition was abolished when the cyclic phosphate was opened by mild acid treatment. The lipid cleaved by phospholipase C digestion, was identified as 1‐O‐hexadecylglycerol by reverse‐phase thin‐layer chromatography. The glycan core was deaminated and chemically labeled by reduction with NaB 3 H 4 . The labeled glycoprotein was exhaustively treated with pronase and dephosphorylated with 50% HF. Although microheterogeneity of the oligosaccharide moiety was apparent, by thin layer chromatography, a main spot coincident with Man(α1–2) Man(α1–6) Man(α1–4) anhydromannitol was shown, consistent with the conserved core structure of all glycosylphosphatidylinositol anchors analysed to date. Copyright © 1993, Wiley Blackwell. All rights reserved
author	Couto, Alicia Susana Muchnik de Lederkremer, Rosa María
author_facet	Couto, Alicia Susana Muchnik de Lederkremer, Rosa María
author_sort	Couto, Alicia Susana
title	The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies
title_short	The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies
title_full	The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies
title_fullStr	The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies
title_full_unstemmed	The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies
title_sort	glycosylphosphatidylinositol anchor of the trypomastigote‐specific tc‐85 glycoprotein from trypanosoma cruzi metabolic‐labeling and structural studies
publishDate	1993
url	https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00142956_v217_n2_p597_COUTO http://hdl.handle.net/20.500.12110/paper_00142956_v217_n2_p597_COUTO
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The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies

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