Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells

Autores
Vilcaes, Aldo Alejandro; Torres Demichelis, Vanina Andrea; Daniotti, Jose Luis
Año de publicación
2011
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Gangliosides are acidic glycosphingolipids that contain sialicacid residues and are expressed in nearly all vertebrate cells.They are synthesized at the Golgi complex by a combination ofglycosyltransferase activities followed by vesicular delivery tothe plasma membrane, where they participate in a variety ofphysiological as well as pathological processes. Recently, a numberof enzymes of ganglioside anabolism and catabolism havebeen shown to be associated with the plasma membrane. In particular,it was observed that CMP-NeuAc:GM3 sialyltransferase(Sial-T2) is able to sialylate GM3 at the plasma membrane (ciscatalyticactivity). In this work, we demonstrated that plasmamembrane-integrated ecto-Sial-T2 also displays a trans-catalyticactivity at the cell surface of epithelial and melanoma cells.By using a highly sensitive enzyme-linked immunosorbent assaycombined with confocal fluorescence microscopy, we observedthat ecto-Sial-T2 was able to sialylate hydrophobically or covalentlyimmobilized GM3 onto a solid surface. More interestingly,we observed that ecto-Sial-T2 was able to sialylate GM3exposed on the membrane of neighboring cells by using both theexogenous and endogenous donor substrate (CMP-N-acetylneuraminicacid) available at the extracellular milieu. In addition,the trans-activity of ecto-Sial-T2 was considerably reducedwhen the expression of the acceptor substrate was inhibited byusing a specific inhibitor of biosynthesis of glycolipids, indicatingthe lipidic nature of the acceptor. Our findings provide thefirst direct evidence that an ecto-sialyltransferase is able totrans-sialylate substrates exposed in the plasma membranefrom mammalian cells, which represents a novel insight into themolecular events that regulate the local glycosphingolipidcomposition.
Fil: Vilcaes, Aldo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Torres Demichelis, Vanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Daniotti, Jose Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Materia
GLYCOSYLTRANSFERASES
GANGLIOSIDES
PLASMA MEMBRANE
ECTO SYALYLTRANSFERASES
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/230262
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/230262
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cellsVilcaes, Aldo AlejandroTorres Demichelis, Vanina AndreaDaniotti, Jose LuisGLYCOSYLTRANSFERASESGANGLIOSIDESPLASMA MEMBRANEECTO SYALYLTRANSFERASEShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Gangliosides are acidic glycosphingolipids that contain sialicacid residues and are expressed in nearly all vertebrate cells.They are synthesized at the Golgi complex by a combination ofglycosyltransferase activities followed by vesicular delivery tothe plasma membrane, where they participate in a variety ofphysiological as well as pathological processes. Recently, a numberof enzymes of ganglioside anabolism and catabolism havebeen shown to be associated with the plasma membrane. In particular,it was observed that CMP-NeuAc:GM3 sialyltransferase(Sial-T2) is able to sialylate GM3 at the plasma membrane (ciscatalyticactivity). In this work, we demonstrated that plasmamembrane-integrated ecto-Sial-T2 also displays a trans-catalyticactivity at the cell surface of epithelial and melanoma cells.By using a highly sensitive enzyme-linked immunosorbent assaycombined with confocal fluorescence microscopy, we observedthat ecto-Sial-T2 was able to sialylate hydrophobically or covalentlyimmobilized GM3 onto a solid surface. More interestingly,we observed that ecto-Sial-T2 was able to sialylate GM3exposed on the membrane of neighboring cells by using both theexogenous and endogenous donor substrate (CMP-N-acetylneuraminicacid) available at the extracellular milieu. In addition,the trans-activity of ecto-Sial-T2 was considerably reducedwhen the expression of the acceptor substrate was inhibited byusing a specific inhibitor of biosynthesis of glycolipids, indicatingthe lipidic nature of the acceptor. Our findings provide thefirst direct evidence that an ecto-sialyltransferase is able totrans-sialylate substrates exposed in the plasma membranefrom mammalian cells, which represents a novel insight into themolecular events that regulate the local glycosphingolipidcomposition.Fil: Vilcaes, Aldo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Torres Demichelis, Vanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaFil: Daniotti, Jose Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; ArgentinaAmerican Society for Biochemistry and Molecular Biology2011-09-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/230262Vilcaes, Aldo Alejandro; Torres Demichelis, Vanina Andrea; Daniotti, Jose Luis; Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 286; 36; 9-9-2011; 31437-314460021-92581083-351XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925820722532info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M111.257196info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:23:23Zoai:ri.conicet.gov.ar:11336/230262instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 10:23:23.29CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
title Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
spellingShingle Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
Vilcaes, Aldo Alejandro
GLYCOSYLTRANSFERASES
GANGLIOSIDES
PLASMA MEMBRANE
ECTO SYALYLTRANSFERASES
title_short Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
title_full Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
title_fullStr Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
title_full_unstemmed Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
title_sort Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells
dc.creator.none.fl_str_mv Vilcaes, Aldo Alejandro
Torres Demichelis, Vanina Andrea
Daniotti, Jose Luis
author Vilcaes, Aldo Alejandro
author_facet Vilcaes, Aldo Alejandro
Torres Demichelis, Vanina Andrea
Daniotti, Jose Luis
author_role author
author2 Torres Demichelis, Vanina Andrea
Daniotti, Jose Luis
author2_role author
author
dc.subject.none.fl_str_mv GLYCOSYLTRANSFERASES
GANGLIOSIDES
PLASMA MEMBRANE
ECTO SYALYLTRANSFERASES
topic GLYCOSYLTRANSFERASES
GANGLIOSIDES
PLASMA MEMBRANE
ECTO SYALYLTRANSFERASES
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Gangliosides are acidic glycosphingolipids that contain sialicacid residues and are expressed in nearly all vertebrate cells.They are synthesized at the Golgi complex by a combination ofglycosyltransferase activities followed by vesicular delivery tothe plasma membrane, where they participate in a variety ofphysiological as well as pathological processes. Recently, a numberof enzymes of ganglioside anabolism and catabolism havebeen shown to be associated with the plasma membrane. In particular,it was observed that CMP-NeuAc:GM3 sialyltransferase(Sial-T2) is able to sialylate GM3 at the plasma membrane (ciscatalyticactivity). In this work, we demonstrated that plasmamembrane-integrated ecto-Sial-T2 also displays a trans-catalyticactivity at the cell surface of epithelial and melanoma cells.By using a highly sensitive enzyme-linked immunosorbent assaycombined with confocal fluorescence microscopy, we observedthat ecto-Sial-T2 was able to sialylate hydrophobically or covalentlyimmobilized GM3 onto a solid surface. More interestingly,we observed that ecto-Sial-T2 was able to sialylate GM3exposed on the membrane of neighboring cells by using both theexogenous and endogenous donor substrate (CMP-N-acetylneuraminicacid) available at the extracellular milieu. In addition,the trans-activity of ecto-Sial-T2 was considerably reducedwhen the expression of the acceptor substrate was inhibited byusing a specific inhibitor of biosynthesis of glycolipids, indicatingthe lipidic nature of the acceptor. Our findings provide thefirst direct evidence that an ecto-sialyltransferase is able totrans-sialylate substrates exposed in the plasma membranefrom mammalian cells, which represents a novel insight into themolecular events that regulate the local glycosphingolipidcomposition.
Fil: Vilcaes, Aldo Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Torres Demichelis, Vanina Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
Fil: Daniotti, Jose Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Centro de Investigaciones en Química Biológica de Córdoba. Universidad Nacional de Córdoba. Facultad de Ciencias Químicas. Centro de Investigaciones en Química Biológica de Córdoba; Argentina
description Gangliosides are acidic glycosphingolipids that contain sialicacid residues and are expressed in nearly all vertebrate cells.They are synthesized at the Golgi complex by a combination ofglycosyltransferase activities followed by vesicular delivery tothe plasma membrane, where they participate in a variety ofphysiological as well as pathological processes. Recently, a numberof enzymes of ganglioside anabolism and catabolism havebeen shown to be associated with the plasma membrane. In particular,it was observed that CMP-NeuAc:GM3 sialyltransferase(Sial-T2) is able to sialylate GM3 at the plasma membrane (ciscatalyticactivity). In this work, we demonstrated that plasmamembrane-integrated ecto-Sial-T2 also displays a trans-catalyticactivity at the cell surface of epithelial and melanoma cells.By using a highly sensitive enzyme-linked immunosorbent assaycombined with confocal fluorescence microscopy, we observedthat ecto-Sial-T2 was able to sialylate hydrophobically or covalentlyimmobilized GM3 onto a solid surface. More interestingly,we observed that ecto-Sial-T2 was able to sialylate GM3exposed on the membrane of neighboring cells by using both theexogenous and endogenous donor substrate (CMP-N-acetylneuraminicacid) available at the extracellular milieu. In addition,the trans-activity of ecto-Sial-T2 was considerably reducedwhen the expression of the acceptor substrate was inhibited byusing a specific inhibitor of biosynthesis of glycolipids, indicatingthe lipidic nature of the acceptor. Our findings provide thefirst direct evidence that an ecto-sialyltransferase is able totrans-sialylate substrates exposed in the plasma membranefrom mammalian cells, which represents a novel insight into themolecular events that regulate the local glycosphingolipidcomposition.
publishDate 2011
dc.date.none.fl_str_mv 2011-09-09
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/230262
Vilcaes, Aldo Alejandro; Torres Demichelis, Vanina Andrea; Daniotti, Jose Luis; Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 286; 36; 9-9-2011; 31437-31446
0021-9258
1083-351X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/230262
identifier_str_mv Vilcaes, Aldo Alejandro; Torres Demichelis, Vanina Andrea; Daniotti, Jose Luis; Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells; American Society for Biochemistry and Molecular Biology; Journal of Biological Chemistry (online); 286; 36; 9-9-2011; 31437-31446
0021-9258
1083-351X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0021925820722532
info:eu-repo/semantics/altIdentifier/doi/10.1074/jbc.M111.257196
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
publisher.none.fl_str_mv American Society for Biochemistry and Molecular Biology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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