New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells

Autores
Bistue Millon, Maria Beatriz; Amorosi, Cyntia Anabel; Papazoglu, Gabriela Magali; Siravegna, Myriam Gladys; Elso, Graciela Raquel; Asteggiano, Carla Gabriela
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Na+/Ca2+ exchangers (NCX and NCKX proteins) contribute to Ca2+ homeostasis and recent studies have demonstrated the expression of NCX1 (SLC8) and NCKX1 (SLC24) proteins in human platelets. A tight ([Ca2+]i) is necessary for platelets to prevent inappropriate thrombus formation or bleeding due to altered platelet aggregation, a severe clinical manifestation in congenital disorders of glycosylation (CDG) PMM2-CDG patients. Very little is known about glycosylation and Ca2+ uptake. In this study, we propose to examine Na+/Ca2+ activity (45-Ca2+ uptake) and protein glycosylation in human cells. Immunopurified NCX1 and NCKX1 proteins from microsomal fractions of human platelets were detected with anti-SLC8 antibody and anti-SLC24 antibody, respectively, and lectin staining (concanavalin A (Con A) and wheat germ agglutinin (WGA)). Additionally, enzymatic N- and O-deglycosylation strategies (PNGase F and O-glycosidase digestion) were assayed. In healthy control subjects, we observed N-linked glycans attached to NCX1 and NCKX1 proteins and O-linked sialo oligosaccharides attached to NCKX1. To better understand the clinical relevance of altered protein N-glycosylation, we analyzed the 45-Ca2+ uptake in PMM2-CDG platelets and transfected NCX1 cDNA in tunicamycin-treated HEK293 cells. Western blot showed that 45-Ca2+ uptake and NCX1 protein were greatly diminished. We present evidence that suggests for the first time that N-hypoglycosylation alters Na+/Ca2+ exchange in platelets from CDG patients or tunicamycin-treated HEK293 cells. Additional studies will be necessary to further elucidate the structure of glycans bound to these proteins and the pathological aspects of cell hypoglycosylation.
Fil: Bistue Millon, Maria Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; Argentina
Fil: Amorosi, Cyntia Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Papazoglu, Gabriela Magali. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; Argentina
Fil: Siravegna, Myriam Gladys. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Elso, Graciela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Asteggiano, Carla Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; Argentina
Materia
Na+/Ca2+ EXCHANGERS
CDG
GLYCOSYLATION
HUMAN PLATELET
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:11336/64405
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/64405
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling New insights about Na+/Ca2+ exchangers and protein glycosylation in human cellsBistue Millon, Maria BeatrizAmorosi, Cyntia AnabelPapazoglu, Gabriela MagaliSiravegna, Myriam GladysElso, Graciela RaquelAsteggiano, Carla GabrielaNa+/Ca2+ EXCHANGERSCDGGLYCOSYLATIONHUMAN PLATELEThttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Na+/Ca2+ exchangers (NCX and NCKX proteins) contribute to Ca2+ homeostasis and recent studies have demonstrated the expression of NCX1 (SLC8) and NCKX1 (SLC24) proteins in human platelets. A tight ([Ca2+]i) is necessary for platelets to prevent inappropriate thrombus formation or bleeding due to altered platelet aggregation, a severe clinical manifestation in congenital disorders of glycosylation (CDG) PMM2-CDG patients. Very little is known about glycosylation and Ca2+ uptake. In this study, we propose to examine Na+/Ca2+ activity (45-Ca2+ uptake) and protein glycosylation in human cells. Immunopurified NCX1 and NCKX1 proteins from microsomal fractions of human platelets were detected with anti-SLC8 antibody and anti-SLC24 antibody, respectively, and lectin staining (concanavalin A (Con A) and wheat germ agglutinin (WGA)). Additionally, enzymatic N- and O-deglycosylation strategies (PNGase F and O-glycosidase digestion) were assayed. In healthy control subjects, we observed N-linked glycans attached to NCX1 and NCKX1 proteins and O-linked sialo oligosaccharides attached to NCKX1. To better understand the clinical relevance of altered protein N-glycosylation, we analyzed the 45-Ca2+ uptake in PMM2-CDG platelets and transfected NCX1 cDNA in tunicamycin-treated HEK293 cells. Western blot showed that 45-Ca2+ uptake and NCX1 protein were greatly diminished. We present evidence that suggests for the first time that N-hypoglycosylation alters Na+/Ca2+ exchange in platelets from CDG patients or tunicamycin-treated HEK293 cells. Additional studies will be necessary to further elucidate the structure of glycans bound to these proteins and the pathological aspects of cell hypoglycosylation.Fil: Bistue Millon, Maria Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; ArgentinaFil: Amorosi, Cyntia Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Papazoglu, Gabriela Magali. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; ArgentinaFil: Siravegna, Myriam Gladys. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Elso, Graciela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Asteggiano, Carla Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; ArgentinaReaserch Trends2017-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/64405Bistue Millon, Maria Beatriz; Amorosi, Cyntia Anabel; Papazoglu, Gabriela Magali; Siravegna, Myriam Gladys; Elso, Graciela Raquel; et al.; New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells; Reaserch Trends; Trends in Cell and Molecular Biology; 12; 9-2017; 17-320972-8449CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.researchtrends.net/tia/article_pdf.asp?in=0&vn=12&tid=55&aid=6060info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:42:15Zoai:ri.conicet.gov.ar:11336/64405instacron: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 09:42:16.089CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
title New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
spellingShingle New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
Bistue Millon, Maria Beatriz
Na+/Ca2+ EXCHANGERS
CDG
GLYCOSYLATION
HUMAN PLATELET
title_short New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
title_full New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
title_fullStr New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
title_full_unstemmed New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
title_sort New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells
dc.creator.none.fl_str_mv Bistue Millon, Maria Beatriz
Amorosi, Cyntia Anabel
Papazoglu, Gabriela Magali
Siravegna, Myriam Gladys
Elso, Graciela Raquel
Asteggiano, Carla Gabriela
author Bistue Millon, Maria Beatriz
author_facet Bistue Millon, Maria Beatriz
Amorosi, Cyntia Anabel
Papazoglu, Gabriela Magali
Siravegna, Myriam Gladys
Elso, Graciela Raquel
Asteggiano, Carla Gabriela
author_role author
author2 Amorosi, Cyntia Anabel
Papazoglu, Gabriela Magali
Siravegna, Myriam Gladys
Elso, Graciela Raquel
Asteggiano, Carla Gabriela
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Na+/Ca2+ EXCHANGERS
CDG
GLYCOSYLATION
HUMAN PLATELET
topic Na+/Ca2+ EXCHANGERS
CDG
GLYCOSYLATION
HUMAN PLATELET
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Na+/Ca2+ exchangers (NCX and NCKX proteins) contribute to Ca2+ homeostasis and recent studies have demonstrated the expression of NCX1 (SLC8) and NCKX1 (SLC24) proteins in human platelets. A tight ([Ca2+]i) is necessary for platelets to prevent inappropriate thrombus formation or bleeding due to altered platelet aggregation, a severe clinical manifestation in congenital disorders of glycosylation (CDG) PMM2-CDG patients. Very little is known about glycosylation and Ca2+ uptake. In this study, we propose to examine Na+/Ca2+ activity (45-Ca2+ uptake) and protein glycosylation in human cells. Immunopurified NCX1 and NCKX1 proteins from microsomal fractions of human platelets were detected with anti-SLC8 antibody and anti-SLC24 antibody, respectively, and lectin staining (concanavalin A (Con A) and wheat germ agglutinin (WGA)). Additionally, enzymatic N- and O-deglycosylation strategies (PNGase F and O-glycosidase digestion) were assayed. In healthy control subjects, we observed N-linked glycans attached to NCX1 and NCKX1 proteins and O-linked sialo oligosaccharides attached to NCKX1. To better understand the clinical relevance of altered protein N-glycosylation, we analyzed the 45-Ca2+ uptake in PMM2-CDG platelets and transfected NCX1 cDNA in tunicamycin-treated HEK293 cells. Western blot showed that 45-Ca2+ uptake and NCX1 protein were greatly diminished. We present evidence that suggests for the first time that N-hypoglycosylation alters Na+/Ca2+ exchange in platelets from CDG patients or tunicamycin-treated HEK293 cells. Additional studies will be necessary to further elucidate the structure of glycans bound to these proteins and the pathological aspects of cell hypoglycosylation.
Fil: Bistue Millon, Maria Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; Argentina
Fil: Amorosi, Cyntia Anabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Papazoglu, Gabriela Magali. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; Argentina
Fil: Siravegna, Myriam Gladys. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Elso, Graciela Raquel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentina
Fil: Asteggiano, Carla Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentina. Universidad Nacional de Córdoba; Argentina
description Na+/Ca2+ exchangers (NCX and NCKX proteins) contribute to Ca2+ homeostasis and recent studies have demonstrated the expression of NCX1 (SLC8) and NCKX1 (SLC24) proteins in human platelets. A tight ([Ca2+]i) is necessary for platelets to prevent inappropriate thrombus formation or bleeding due to altered platelet aggregation, a severe clinical manifestation in congenital disorders of glycosylation (CDG) PMM2-CDG patients. Very little is known about glycosylation and Ca2+ uptake. In this study, we propose to examine Na+/Ca2+ activity (45-Ca2+ uptake) and protein glycosylation in human cells. Immunopurified NCX1 and NCKX1 proteins from microsomal fractions of human platelets were detected with anti-SLC8 antibody and anti-SLC24 antibody, respectively, and lectin staining (concanavalin A (Con A) and wheat germ agglutinin (WGA)). Additionally, enzymatic N- and O-deglycosylation strategies (PNGase F and O-glycosidase digestion) were assayed. In healthy control subjects, we observed N-linked glycans attached to NCX1 and NCKX1 proteins and O-linked sialo oligosaccharides attached to NCKX1. To better understand the clinical relevance of altered protein N-glycosylation, we analyzed the 45-Ca2+ uptake in PMM2-CDG platelets and transfected NCX1 cDNA in tunicamycin-treated HEK293 cells. Western blot showed that 45-Ca2+ uptake and NCX1 protein were greatly diminished. We present evidence that suggests for the first time that N-hypoglycosylation alters Na+/Ca2+ exchange in platelets from CDG patients or tunicamycin-treated HEK293 cells. Additional studies will be necessary to further elucidate the structure of glycans bound to these proteins and the pathological aspects of cell hypoglycosylation.
publishDate 2017
dc.date.none.fl_str_mv 2017-09
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/11336/64405
Bistue Millon, Maria Beatriz; Amorosi, Cyntia Anabel; Papazoglu, Gabriela Magali; Siravegna, Myriam Gladys; Elso, Graciela Raquel; et al.; New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells; Reaserch Trends; Trends in Cell and Molecular Biology; 12; 9-2017; 17-32
0972-8449
CONICET Digital
CONICET
url http://hdl.handle.net/11336/64405
identifier_str_mv Bistue Millon, Maria Beatriz; Amorosi, Cyntia Anabel; Papazoglu, Gabriela Magali; Siravegna, Myriam Gladys; Elso, Graciela Raquel; et al.; New insights about Na+/Ca2+ exchangers and protein glycosylation in human cells; Reaserch Trends; Trends in Cell and Molecular Biology; 12; 9-2017; 17-32
0972-8449
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.researchtrends.net/tia/article_pdf.asp?in=0&vn=12&tid=55&aid=6060
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
dc.publisher.none.fl_str_mv Reaserch Trends
publisher.none.fl_str_mv Reaserch Trends
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 13.070432

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