Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II

Autores
Ruggiero, Fernando Miguel; Vilcaes, Aldo Alejandro; Iglesias Bartolome, Ramiro; Daniotti, Jose Luis
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. It contains two putative N-glycosylation sites (Asn92 and Asn211). Whereas Asn92 is only conserved in mammalian species, Asn211 is highly conserved in mammals, birds and fish. The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. We found that ST3Gal-II distributes along the Golgi complex, mainly in proximal compartments. By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn211 and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. Whereas the N-glycan at position Asn211 seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn92 had only a moderate effect. Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme.
Fil: Ruggiero, Fernando Miguel. 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: 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: Iglesias Bartolome, Ramiro. National Institutes of Health; Estados Unidos
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
Ganglioside
Glycolipid
Golgi Complex
N-Glycan Trimming
N-Glycosylation
Sialyltransferase
St3gal-Ii
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/50863
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/50863
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-IIRuggiero, Fernando MiguelVilcaes, Aldo AlejandroIglesias Bartolome, RamiroDaniotti, Jose LuisGangliosideGlycolipidGolgi ComplexN-Glycan TrimmingN-GlycosylationSialyltransferaseSt3gal-Iihttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. It contains two putative N-glycosylation sites (Asn92 and Asn211). Whereas Asn92 is only conserved in mammalian species, Asn211 is highly conserved in mammals, birds and fish. The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. We found that ST3Gal-II distributes along the Golgi complex, mainly in proximal compartments. By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn211 and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. Whereas the N-glycan at position Asn211 seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn92 had only a moderate effect. Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme.Fil: Ruggiero, Fernando Miguel. 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: 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: Iglesias Bartolome, Ramiro. National Institutes of Health; Estados UnidosFil: 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; ArgentinaPortland Press2015-07info: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/11336/50863Ruggiero, Fernando Miguel; Vilcaes, Aldo Alejandro; Iglesias Bartolome, Ramiro; Daniotti, Jose Luis; Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II; Portland Press; Biochemical Journal; 469; 1; 7-2015; 83-950264-60211470-8728CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.biochemj.org/content/469/1/83info:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20150072info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:00:31Zoai:ri.conicet.gov.ar:11336/50863instacron: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:34982025-09-03 10:00:31.98CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
title Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
spellingShingle Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
Ruggiero, Fernando Miguel
Ganglioside
Glycolipid
Golgi Complex
N-Glycan Trimming
N-Glycosylation
Sialyltransferase
St3gal-Ii
title_short Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
title_full Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
title_fullStr Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
title_full_unstemmed Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
title_sort Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II
dc.creator.none.fl_str_mv Ruggiero, Fernando Miguel
Vilcaes, Aldo Alejandro
Iglesias Bartolome, Ramiro
Daniotti, Jose Luis
author Ruggiero, Fernando Miguel
author_facet Ruggiero, Fernando Miguel
Vilcaes, Aldo Alejandro
Iglesias Bartolome, Ramiro
Daniotti, Jose Luis
author_role author
author2 Vilcaes, Aldo Alejandro
Iglesias Bartolome, Ramiro
Daniotti, Jose Luis
author2_role author
author
author
dc.subject.none.fl_str_mv Ganglioside
Glycolipid
Golgi Complex
N-Glycan Trimming
N-Glycosylation
Sialyltransferase
St3gal-Ii
topic Ganglioside
Glycolipid
Golgi Complex
N-Glycan Trimming
N-Glycosylation
Sialyltransferase
St3gal-Ii
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. It contains two putative N-glycosylation sites (Asn92 and Asn211). Whereas Asn92 is only conserved in mammalian species, Asn211 is highly conserved in mammals, birds and fish. The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. We found that ST3Gal-II distributes along the Golgi complex, mainly in proximal compartments. By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn211 and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. Whereas the N-glycan at position Asn211 seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn92 had only a moderate effect. Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme.
Fil: Ruggiero, Fernando Miguel. 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: 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: Iglesias Bartolome, Ramiro. National Institutes of Health; Estados Unidos
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 ST3Gal-II, a type II transmembrane protein, is the main mammalian sialyltransferase responsible for GD1a and GT1b ganglioside biosynthesis in brain. It contains two putative N-glycosylation sites (Asn92 and Asn211). Whereas Asn92 is only conserved in mammalian species, Asn211 is highly conserved in mammals, birds and fish. The present study explores the occupancy and relevance for intracellular trafficking and enzyme activity of these potential N-glycosylations in human ST3Gal-II. We found that ST3Gal-II distributes along the Golgi complex, mainly in proximal compartments. By pharmacological, biochemical and site-directed mutagenesis, we observed that ST3Gal-II is mostly N-glycosylated at Asn211 and that this co-translational modification is critical for its exit from the endoplasmic reticulum and proper Golgi localization. The individual N-glycosylation sites had different effects on ST3Gal-II enzymatic activity. Whereas the N-glycan at position Asn211 seems to negatively influence the activity of the enzyme using both glycolipid and glycoprotein as acceptor substrates, the single N-glycan mutant at Asn92 had only a moderate effect. Lastly, we demonstrated that the N-terminal ST3Gal-II domain containing the cytosolic, transmembrane and stem region (amino acids 1-51) is able to drive a protein reporter out of the endoplasmic reticulum and to retain it in the Golgi complex. This suggests that the C-terminal domain of ST3Gal-II depends on N-glycosylation to attain an optimum conformation for proper exit from the endoplasmic reticulum, but it does not represent an absolute requirement for Golgi complex retention of the enzyme.
publishDate 2015
dc.date.none.fl_str_mv 2015-07
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/11336/50863
Ruggiero, Fernando Miguel; Vilcaes, Aldo Alejandro; Iglesias Bartolome, Ramiro; Daniotti, Jose Luis; Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II; Portland Press; Biochemical Journal; 469; 1; 7-2015; 83-95
0264-6021
1470-8728
CONICET Digital
CONICET
url http://hdl.handle.net/11336/50863
identifier_str_mv Ruggiero, Fernando Miguel; Vilcaes, Aldo Alejandro; Iglesias Bartolome, Ramiro; Daniotti, Jose Luis; Critical role of evolutionarily conserved glycosylation at Asn211in the intracellular trafficking and activity of sialyltransferase ST3Gal-II; Portland Press; Biochemical Journal; 469; 1; 7-2015; 83-95
0264-6021
1470-8728
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.biochemj.org/content/469/1/83
info:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20150072
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Portland Press
publisher.none.fl_str_mv Portland Press
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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score 13.13397

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