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Abstract:

Both, culture-derived and metacyclic trypomastigotes of Trypanosoma cruzi shed a glycoprotein, the shed acute phase antigen, that is responsible for the trans-sialidase activity. In the present work the structure of the glycosylphosphatidylinositol membrane anchor of the trans-sialidase isolated from metacyclic forms was determined. Parasites were metabolically labelled with [9, 10(n)3H]-palmitic acid and the glycoprotein was purified by immunoprecipitation with a monoclonal antibody directed against the repetitive aminoacid sequence. Treatment of the glycoprotein with phosphatidylinositol phospholipase C (PI-PLC) from Bacillus thuringiensis rendered a lipid that comigrated in TLC with a standard of ceramide. No alkylglycerol was detected in contrast with the results previously obtained for the trans-sialidase isolated from culture-derived trypomastigotes where both lipids were found. Chemical and chromatographic analysis showed that the lipid moiety is palmitoyldihydrosphingosine with a minor amount of stearoyldihydrosphingosine. The glycan constituent of the glycosylphosphatidylinositol-anchor was analysed by nitrous acid deamination of the aqueous phase of the PI-PLC treatment, followed by reduction with NaBH4 and hydrolysis of the phosphodiester with aqueous hydrofluoric acid. A major oligosaccharide was obtained and enzymatic treatment with exoglycosidases and further chromatography in a high pH anion exchange system showed that the trimannosyl core backbone is substituted by an α-galactose. A comparison between the lipid constituent of the glycosylphosphatidylinositol anchor of several proteins and their spontaneous shedding by the action of an endogenous PI-PLC was made. Copyright (C) 1998 Elsevier Science B.V.

Registro:

Documento: Artículo
Título:Structure of the glycosylphosphatidylinositol-anchor of the trans-sialidase from Trypanosoma cruzi metacyclic trypomastigote forms
Autor:Agusti, R.; Couto, A.S.; Campetella, O.; Frasch, A.C.C.; De Lederkremer, R.M.
Filiación:Depto. de Quím. Orgán., Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina
Inst. de Invest. Biotecnologicas, Univ. Nac. de Gral. San Martin, Buenos Aires, Argentina
Depto. de Quím. Orgán., Pabellón 2, Ciudad Universitaria, 1428 Buenos Aires, Argentina
Palabras clave:Ceramide; GPI anchor; Trans-sialidase; Trypanosoma cruzi; ceramide; glycosylphosphatidylinositol; membrane protein; sialidase; anion exchange chromatography; article; controlled study; enzyme activity; nonhuman; priority journal; thin layer chromatography; Trypanosoma cruzi; trypomastigote; Bacillus thuringiensis; Trypanosoma; Trypanosoma cruzi
Año:1998
Volumen:97
Número:1-2
Página de inicio:123
Página de fin:131
DOI: http://dx.doi.org/10.1016/S0166-6851(98)00137-6
Título revista:Molecular and Biochemical Parasitology
Título revista abreviado:Mol. Biochem. Parasitol.
ISSN:01666851
CODEN:MBIPD
CAS:sialidase, 9001-67-6
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01666851_v97_n1-2_p123_Agusti

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Citas:

---------- APA ----------
Agusti, R., Couto, A.S., Campetella, O., Frasch, A.C.C. & De Lederkremer, R.M. (1998) . Structure of the glycosylphosphatidylinositol-anchor of the trans-sialidase from Trypanosoma cruzi metacyclic trypomastigote forms. Molecular and Biochemical Parasitology, 97(1-2), 123-131.
http://dx.doi.org/10.1016/S0166-6851(98)00137-6
---------- CHICAGO ----------
Agusti, R., Couto, A.S., Campetella, O., Frasch, A.C.C., De Lederkremer, R.M. "Structure of the glycosylphosphatidylinositol-anchor of the trans-sialidase from Trypanosoma cruzi metacyclic trypomastigote forms" . Molecular and Biochemical Parasitology 97, no. 1-2 (1998) : 123-131.
http://dx.doi.org/10.1016/S0166-6851(98)00137-6
---------- MLA ----------
Agusti, R., Couto, A.S., Campetella, O., Frasch, A.C.C., De Lederkremer, R.M. "Structure of the glycosylphosphatidylinositol-anchor of the trans-sialidase from Trypanosoma cruzi metacyclic trypomastigote forms" . Molecular and Biochemical Parasitology, vol. 97, no. 1-2, 1998, pp. 123-131.
http://dx.doi.org/10.1016/S0166-6851(98)00137-6
---------- VANCOUVER ----------
Agusti, R., Couto, A.S., Campetella, O., Frasch, A.C.C., De Lederkremer, R.M. Structure of the glycosylphosphatidylinositol-anchor of the trans-sialidase from Trypanosoma cruzi metacyclic trypomastigote forms. Mol. Biochem. Parasitol. 1998;97(1-2):123-131.
http://dx.doi.org/10.1016/S0166-6851(98)00137-6