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
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/132362
Ver los metadatos del registro completo
id |
CONICETDig_6178fc121d3a63a7d6f896a1b33360a5 |
---|---|
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 Técnicas2025-09-29T09:56:51Zoai: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:34982025-09-29 09:56:51.459CONICET 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 |
_version_ |
1844613705088630784 |
score |
13.070432 |