Abstract:
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
Registro:
Documento: |
Artículo
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Título: | The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies |
Autor: | COUTO, A.S.; DE LEDERKREMER, R.M.; COLLI, W.; ALVES, M.J.M. |
Filiación: | Departamento de Química Orgánica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Argentina Departamento de Bioquímica, Instituto de Quimica, Universidade de Sao Paulo, Brazil
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Palabras clave: | 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 |
Año: | 1993
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Volumen: | 217
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Número: | 2
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Página de inicio: | 597
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Página de fin: | 602
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DOI: |
http://dx.doi.org/10.1111/j.1432-1033.1993.tb18282.x |
Título revista: | European Journal of Biochemistry
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Título revista abreviado: | Eur. J. Biochem.
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ISSN: | 00142956
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CAS: | Antibodies, Monoclonal; Glycoproteins; Glycosylphosphatidylinositols; Lipids; Palmitic Acid, 57-10-3; Palmitic Acids; Protozoan Proteins; Wheat Germ Agglutinins
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Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00142956_v217_n2_p597_COUTO |
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Citas:
---------- APA ----------
COUTO, A.S., DE LEDERKREMER, R.M., COLLI, W. & ALVES, M.J.M.
(1993)
. The glycosylphosphatidylinositol anchor of the trypomastigote‐specific Tc‐85 glycoprotein from Trypanosoma cruzi Metabolic‐labeling and structural studies. European Journal of Biochemistry, 217(2), 597-602.
http://dx.doi.org/10.1111/j.1432-1033.1993.tb18282.x---------- CHICAGO ----------
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"
. European Journal of Biochemistry 217, no. 2
(1993) : 597-602.
http://dx.doi.org/10.1111/j.1432-1033.1993.tb18282.x---------- MLA ----------
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"
. European Journal of Biochemistry, vol. 217, no. 2, 1993, pp. 597-602.
http://dx.doi.org/10.1111/j.1432-1033.1993.tb18282.x---------- VANCOUVER ----------
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. Eur. J. Biochem. 1993;217(2):597-602.
http://dx.doi.org/10.1111/j.1432-1033.1993.tb18282.x