uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition

Autores
Sahores, Maria Macarena; Prinetti, Alessandro; Chiabrando, Gustavo Alberto; Blasi, Francesco; Sonnino, Sandro
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
UPAR is a GPI anchored protein, which is found in both lipid rafts and in more fluid regions of the plasma membrane. We have studied the role of the ligand uPA on uPAR localization and on the composition of the lipid membrane microdomains. We have analyzed the glycosphingolipid environment of uPAR in detergent resistant membrane (DRM) fractions prepared by cell lysis with 1% Triton X-100 and fractionated by sucrose gradient centrifugation obtained from HEK293-uPAR cells. The uPAR specific lipid membrane microdomain has been separated from the total DRM fraction by immunoprecipitation with an anti-uPAR specific antibody under conditions that preserve membrane integrity. We have also tested uPA-induced ERK phosphorylation in the presence of methyl-β-cyclodextrin, which is known to disrupt lipid rafts by sequestering cholesterol from such domains. Our results show that uPAR is partially associated with DRM and this association is increased by ligands, is independent of the catalytic activity of uPA, and is required for intracellular signalling. In the absence of ligands, uPAR experiences a lipid environment very similar to that of total DRM, enriched in sphingomyelin and glycosphingolipids. However, after treatment of cells with uPA or ATF the lipid environment is strongly impoverished of neutral glycosphingolipids. © 2007.
Fil: Sahores, Maria Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Prinetti, Alessandro. Università degli Studi di Milano; Italia
Fil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Blasi, Francesco. FIRC Institute of Molecular Oncology; Italia. Università Vita Salute San Raffaele; Italia
Fil: Sonnino, Sandro. Università degli Studi di Milano; Italia
Materia
IMMUNOPRECIPITATION
LIPID RAFT
UPA RECEPTOR
UROKINASE
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:11336/132362
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/132362
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain compositionSahores, Maria MacarenaPrinetti, AlessandroChiabrando, Gustavo AlbertoBlasi, FrancescoSonnino, SandroIMMUNOPRECIPITATIONLIPID RAFTUPA RECEPTORUROKINASEhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1UPAR is a GPI anchored protein, which is found in both lipid rafts and in more fluid regions of the plasma membrane. We have studied the role of the ligand uPA on uPAR localization and on the composition of the lipid membrane microdomains. We have analyzed the glycosphingolipid environment of uPAR in detergent resistant membrane (DRM) fractions prepared by cell lysis with 1% Triton X-100 and fractionated by sucrose gradient centrifugation obtained from HEK293-uPAR cells. The uPAR specific lipid membrane microdomain has been separated from the total DRM fraction by immunoprecipitation with an anti-uPAR specific antibody under conditions that preserve membrane integrity. We have also tested uPA-induced ERK phosphorylation in the presence of methyl-β-cyclodextrin, which is known to disrupt lipid rafts by sequestering cholesterol from such domains. Our results show that uPAR is partially associated with DRM and this association is increased by ligands, is independent of the catalytic activity of uPA, and is required for intracellular signalling. In the absence of ligands, uPAR experiences a lipid environment very similar to that of total DRM, enriched in sphingomyelin and glycosphingolipids. However, after treatment of cells with uPA or ATF the lipid environment is strongly impoverished of neutral glycosphingolipids. © 2007.Fil: Sahores, Maria Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Prinetti, Alessandro. Università degli Studi di Milano; ItaliaFil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; ArgentinaFil: Blasi, Francesco. FIRC Institute of Molecular Oncology; Italia. Università Vita Salute San Raffaele; ItaliaFil: Sonnino, Sandro. Università degli Studi di Milano; ItaliaElsevier Science2008-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/132362Sahores, Maria Macarena; Prinetti, Alessandro; Chiabrando, Gustavo Alberto; Blasi, Francesco; Sonnino, Sandro; uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1778; 1; 1-2008; 250-2590005-2736CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2007.09.030info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0005273607003896info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:26:04Zoai:ri.conicet.gov.ar:11336/132362instacron: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:34982025-10-22 11:26:04.393CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
title uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
spellingShingle uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
Sahores, Maria Macarena
IMMUNOPRECIPITATION
LIPID RAFT
UPA RECEPTOR
UROKINASE
title_short uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
title_full uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
title_fullStr uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
title_full_unstemmed uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
title_sort uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition
dc.creator.none.fl_str_mv Sahores, Maria Macarena
Prinetti, Alessandro
Chiabrando, Gustavo Alberto
Blasi, Francesco
Sonnino, Sandro
author Sahores, Maria Macarena
author_facet Sahores, Maria Macarena
Prinetti, Alessandro
Chiabrando, Gustavo Alberto
Blasi, Francesco
Sonnino, Sandro
author_role author
author2 Prinetti, Alessandro
Chiabrando, Gustavo Alberto
Blasi, Francesco
Sonnino, Sandro
author2_role author
author
author
author
dc.subject.none.fl_str_mv IMMUNOPRECIPITATION
LIPID RAFT
UPA RECEPTOR
UROKINASE
topic IMMUNOPRECIPITATION
LIPID RAFT
UPA RECEPTOR
UROKINASE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv UPAR is a GPI anchored protein, which is found in both lipid rafts and in more fluid regions of the plasma membrane. We have studied the role of the ligand uPA on uPAR localization and on the composition of the lipid membrane microdomains. We have analyzed the glycosphingolipid environment of uPAR in detergent resistant membrane (DRM) fractions prepared by cell lysis with 1% Triton X-100 and fractionated by sucrose gradient centrifugation obtained from HEK293-uPAR cells. The uPAR specific lipid membrane microdomain has been separated from the total DRM fraction by immunoprecipitation with an anti-uPAR specific antibody under conditions that preserve membrane integrity. We have also tested uPA-induced ERK phosphorylation in the presence of methyl-β-cyclodextrin, which is known to disrupt lipid rafts by sequestering cholesterol from such domains. Our results show that uPAR is partially associated with DRM and this association is increased by ligands, is independent of the catalytic activity of uPA, and is required for intracellular signalling. In the absence of ligands, uPAR experiences a lipid environment very similar to that of total DRM, enriched in sphingomyelin and glycosphingolipids. However, after treatment of cells with uPA or ATF the lipid environment is strongly impoverished of neutral glycosphingolipids. © 2007.
Fil: Sahores, Maria Macarena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Prinetti, Alessandro. Università degli Studi di Milano; Italia
Fil: Chiabrando, Gustavo Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Córdoba. Centro de Investigaciones en Bioquímica Clínica e Inmunología; Argentina
Fil: Blasi, Francesco. FIRC Institute of Molecular Oncology; Italia. Università Vita Salute San Raffaele; Italia
Fil: Sonnino, Sandro. Università degli Studi di Milano; Italia
description UPAR is a GPI anchored protein, which is found in both lipid rafts and in more fluid regions of the plasma membrane. We have studied the role of the ligand uPA on uPAR localization and on the composition of the lipid membrane microdomains. We have analyzed the glycosphingolipid environment of uPAR in detergent resistant membrane (DRM) fractions prepared by cell lysis with 1% Triton X-100 and fractionated by sucrose gradient centrifugation obtained from HEK293-uPAR cells. The uPAR specific lipid membrane microdomain has been separated from the total DRM fraction by immunoprecipitation with an anti-uPAR specific antibody under conditions that preserve membrane integrity. We have also tested uPA-induced ERK phosphorylation in the presence of methyl-β-cyclodextrin, which is known to disrupt lipid rafts by sequestering cholesterol from such domains. Our results show that uPAR is partially associated with DRM and this association is increased by ligands, is independent of the catalytic activity of uPA, and is required for intracellular signalling. In the absence of ligands, uPAR experiences a lipid environment very similar to that of total DRM, enriched in sphingomyelin and glycosphingolipids. However, after treatment of cells with uPA or ATF the lipid environment is strongly impoverished of neutral glycosphingolipids. © 2007.
publishDate 2008
dc.date.none.fl_str_mv 2008-01
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/11336/132362
Sahores, Maria Macarena; Prinetti, Alessandro; Chiabrando, Gustavo Alberto; Blasi, Francesco; Sonnino, Sandro; uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1778; 1; 1-2008; 250-259
0005-2736
CONICET Digital
CONICET
url http://hdl.handle.net/11336/132362
identifier_str_mv Sahores, Maria Macarena; Prinetti, Alessandro; Chiabrando, Gustavo Alberto; Blasi, Francesco; Sonnino, Sandro; uPA binding increases UPAR localization to lipid rafts and modifies the receptor microdomain composition; Elsevier Science; Biochimica et Biophysica Acta - Biomembranes; 1778; 1; 1-2008; 250-259
0005-2736
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2007.09.030
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0005273607003896
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 Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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 12.982451

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