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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todo:paper_00142956_v217_n2_p597_COUTO2023-10-03T14:11:53Z The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies COUTO, A.S. DE LEDERKREMER, R.M. COLLI, W. ALVES, M.J.M. 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar 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, A.S. DE LEDERKREMER, R.M. COLLI, W. ALVES, M.J.M. 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 |
| format |
JOUR |
| author |
COUTO, A.S. DE LEDERKREMER, R.M. COLLI, W. ALVES, M.J.M. |
| author_facet |
COUTO, A.S. DE LEDERKREMER, R.M. COLLI, W. ALVES, M.J.M. |
| author_sort |
COUTO, A.S. |
| 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 |
| url |
http://hdl.handle.net/20.500.12110/paper_00142956_v217_n2_p597_COUTO |
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