Generation of a soluble form of human endoglin fused to green fluorescent protein

13 p.-5 fig.

Detalles Bibliográficos
Autores: Ruiz-Llorente, Lidia, Vega, María Cristina, Fernández, Francisco J., Langa, Carmen, Morrell, Nicholas W., Upton, Paul D., Bernabéu, Carmelo
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2021
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/262520
Acceso en línea:http://hdl.handle.net/10261/262520
Access Level:acceso abierto
Palabra clave:Endoglin
GFP
Soluble endoglin
Fusion protein
Recombinant protein
Fluorescence
BMP
TGF-β
Endothelium
id ES_75db1d0e092ea3d730a72bc29d3e65b7
oai_identifier_str oai:digital.csic.es:10261/262520
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Generation of a soluble form of human endoglin fused to green fluorescent protein
title Generation of a soluble form of human endoglin fused to green fluorescent protein
spellingShingle Generation of a soluble form of human endoglin fused to green fluorescent protein
Ruiz-Llorente, Lidia
Endoglin
GFP
Soluble endoglin
Fusion protein
Recombinant protein
Fluorescence
BMP
TGF-β
Endothelium
title_short Generation of a soluble form of human endoglin fused to green fluorescent protein
title_full Generation of a soluble form of human endoglin fused to green fluorescent protein
title_fullStr Generation of a soluble form of human endoglin fused to green fluorescent protein
title_full_unstemmed Generation of a soluble form of human endoglin fused to green fluorescent protein
title_sort Generation of a soluble form of human endoglin fused to green fluorescent protein
dc.creator.none.fl_str_mv Ruiz-Llorente, Lidia
Vega, María Cristina
Fernández, Francisco J.
Langa, Carmen
Morrell, Nicholas W.
Upton, Paul D.
Bernabéu, Carmelo
author Ruiz-Llorente, Lidia
author_facet Ruiz-Llorente, Lidia
Vega, María Cristina
Fernández, Francisco J.
Langa, Carmen
Morrell, Nicholas W.
Upton, Paul D.
Bernabéu, Carmelo
author_role author
author2 Vega, María Cristina
Fernández, Francisco J.
Langa, Carmen
Morrell, Nicholas W.
Upton, Paul D.
Bernabéu, Carmelo
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Ciencia, Innovación y Universidades (España)
Centro de Investigación Biomédica en Red Enfermedades Raras (España)
Consejo Superior de Investigaciones Científicas (España)
British Heart Foundation
Instituto de Salud Carlos III
European Commission
Ruiz-Llorente, Lidia [0000-0003-1430-9618]
Vega, María Cristina [0000-0003-0628-8378]
Fernández, Francisco J. [0000-0002-5015-1849]
Morrell, Nicholas W. [0000-0001-5700-9792]
Upton, Paul D. [0000-0003-2716-4921]
Bernabéu, Carmelo [0000-0002-1563-6162]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Endoglin
GFP
Soluble endoglin
Fusion protein
Recombinant protein
Fluorescence
BMP
TGF-β
Endothelium
topic Endoglin
GFP
Soluble endoglin
Fusion protein
Recombinant protein
Fluorescence
BMP
TGF-β
Endothelium
description 13 p.-5 fig.
publishDate 2021
dc.date.none.fl_str_mv 2021
2022
2022
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/262520
url http://hdl.handle.net/10261/262520
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO//SAF2013-43421-R
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-102242-B-I00
https://doi.org/10.3390/ijms222011282
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
publisher.none.fl_str_mv Multidisciplinary Digital Publishing Institute
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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spelling Generation of a soluble form of human endoglin fused to green fluorescent proteinRuiz-Llorente, LidiaVega, María CristinaFernández, Francisco J.Langa, CarmenMorrell, Nicholas W.Upton, Paul D.Bernabéu, CarmeloEndoglinGFPSoluble endoglinFusion proteinRecombinant proteinFluorescenceBMPTGF-βEndothelium13 p.-5 fig.Endoglin (Eng, CD105) is a type I membrane glycoprotein that functions in endothelial cells as an auxiliary receptor for transforming growth factor β (TGF-β)/bone morphogenetic protein (BMP) family members and as an integrin ligand, modulating the vascular pathophysiology. Besides the membrane-bound endoglin, there is a soluble form of endoglin (sEng) that can be generated by the action of the matrix metalloproteinase (MMP)-14 or -12 on the juxtamembrane region of its ectodomain. High levels of sEng have been reported in patients with preeclampsia, hypercholesterolemia, atherosclerosis and cancer. In addition, sEng is a marker of cardiovascular damage in patients with hypertension and diabetes, plays a pathogenic role in preeclampsia, and inhibits angiogenesis and tumor proliferation, migration, and invasion in cancer. However, the mechanisms of action of sEng have not yet been elucidated, and new tools and experimental approaches are necessary to advance in this field. To this end, we aimed to obtain a fluorescent form of sEng as a new tool for biological imaging. Thus, we cloned the extracellular domain of endoglin in the pEGFP-N1 plasmid to generate a fusion protein with green fluorescent protein (GFP), giving rise to pEGFP-N1/Eng.EC. The recombinant fusion protein was characterized by transient and stable transfections in CHO-K1 cells using fluorescence microscopy, SDS-PAGE, immunodetection, and ELISA techniques. Upon transfection with pEGFP-N1/Eng.EC, fluorescence was readily detected in cells, indicating that the GFP contained in the recombinant protein was properly folded into the cytosol. Furthermore, as evidenced by Western blot analysis, the secreted fusion protein yielded the expected molecular mass and displayed a specific fluorescent signal. The fusion protein was also able to bind to BMP9 and BMP10 in vitro. Therefore, the construct described here could be used as a tool for functional in vitro studies of the extracellular domain of endoglin.This research was funded by grants from Ministerio de Ciencia, Innovación y Universidades of Spain (SAF2013-43421-R to C.B. and RTI2018-102242-B-I00 to M.C.V.), Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER; ISCIII-CB06/07/0038 to C.B. and contract CNV-234-PRF-360 to L.R.-L.) and Consejo Superior de Investigaciones Científicas (CSIC; 201920E022 to C.B.) and the British Heart Foundation (Programme Grant RG/19/3/34265 to P.D.U. and N.W.M.). CIBERER is an initiative of the Instituto de Salud Carlos III (ISCIII) of Spain supported by FEDER funds.Peer reviewedMultidisciplinary Digital Publishing InstituteMinisterio de Ciencia, Innovación y Universidades (España)Centro de Investigación Biomédica en Red Enfermedades Raras (España)Consejo Superior de Investigaciones Científicas (España)British Heart FoundationInstituto de Salud Carlos IIIEuropean CommissionRuiz-Llorente, Lidia [0000-0003-1430-9618]Vega, María Cristina [0000-0003-0628-8378]Fernández, Francisco J. [0000-0002-5015-1849]Morrell, Nicholas W. [0000-0001-5700-9792]Upton, Paul D. [0000-0003-2716-4921]Bernabéu, Carmelo [0000-0002-1563-6162]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202220222021info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/262520reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO//SAF2013-43421-Rinfo:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/RTI2018-102242-B-I00https://doi.org/10.3390/ijms222011282Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2625202026-05-22T06:33:51Z
score 15,81155

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