Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc

Heteromeric amino acid transporters (HATs) are the unique example, known in all kingdoms of life, of solute transporters composed of two subunits linked by a conserved disulfide bridge. In metazoans, the heavy subunit is responsible for the trafficking of the heterodimer to the plasma membrane, and...

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Autores: Rosell, Albert, Meury, Marcel, Alvarez-Marimon, Elena, Costa, Meritxell, Pérez-Cano, Laura, Zorzano, Antonio, Fernández-Recio, Juan, Palacín, Manuel, Fotiadis, Dimitrios
Tipo de recurso: artículo
Fecha de publicación:2014
País:España
Institución:Universitat Politècnica de Catalunya (UPC)
Repositorio:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglés
OAI Identifier:oai:upcommons.upc.edu:2117/103456
Acceso en línea:https://hdl.handle.net/2117/103456
https://dx.doi.org/10.1073/pnas.1323779111
Access Level:acceso abierto
Palabra clave:Cellular & molecular mechanisms of toxin action
Amino acids--Analysis
CD98hc
4F2hc ectodomain
Heteromeric amino acid transporters (HATs)
Aminoàcids--Anàlisi
Biologia molecular
Àrees temàtiques de la UPC::Enginyeria biomèdica
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spelling Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hcRosell, AlbertMeury, MarcelAlvarez-Marimon, ElenaCosta, MeritxellPérez-Cano, LauraZorzano, AntonioFernández-Recio, JuanPalacín, ManuelFotiadis, DimitriosCellular & molecular mechanisms of toxin actionAmino acids--AnalysisCD98hc4F2hc ectodomainHeteromeric amino acid transporters (HATs)Aminoàcids--AnàlisiBiologia molecularÀrees temàtiques de la UPC::Enginyeria biomèdicaHeteromeric amino acid transporters (HATs) are the unique example, known in all kingdoms of life, of solute transporters composed of two subunits linked by a conserved disulfide bridge. In metazoans, the heavy subunit is responsible for the trafficking of the heterodimer to the plasma membrane, and the light subunit is the transporter. HATs are involved in human pathologies such as amino acidurias, tumor growth and invasion, viral infection and cocaine addiction. However structural information about interactions between the heavy and light subunits of HATs is scarce. In this work, transmission electron microscopy and single-particle analysis of purified human 4F2hc/L-type amino acid transporter 2 (LAT2) heterodimers overexpressed in the yeast Pichia pastoris, together with docking analysis and crosslinking experiments, reveal that the extracellular domain of 4F2hc interacts with LAT2, almost completely covering the extracellular face of the transporter. 4F2hc increases the stability of the light subunit LAT2 in detergent-solubilized Pichia membranes, allowing functional reconstitution of the heterodimer into proteoliposomes. Moreover, the extracellular domain of 4F2hc suffices to stabilize solubilized LAT2. The interaction of 4F2hc with LAT2 gives insights into the structural bases for light subunit recognition and the stabilizing role of the ancillary protein in HATs.This work was supported by Spanish Ministry of Science and Innovation Grants BIO2010-22324 (to J.F.-R.) and SAF2012- 40080-C02-01, European Commission Frame Program 7 Grant 201924 (European Drug Initative on Channels and Transporters), Fundación Ramón Areces, and the Generalitat de Catalunya Grant SGR2009-1355 (to M.P.); by University of Bern, Swiss National Science Foundation Grants 31003A_125150 and 31003A_144168; the Bern University Research Foundation; the Novartis Foundation; the Marie Curie Actions International Fellowship Program; and National Center of Competence in Research TransCure (D.F.).Peer ReviewedNational Academy of Sciences20142014-02-2520172017-04-07journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/2117/103456https://dx.doi.org/10.1073/pnas.1323779111reponame:UPCommons. Portal del coneixement obert de la UPCinstname:Universitat Politècnica de Catalunya (UPC)InglésengMinisterio de Ciencia e Innovación http://doi.org/10.13039/501100004837 BIO2010-22324 DOCKING ENTRE PROTEINAS Y OTROS ASPECTOS: AVANCES EN LA DESCRIPCION BIOFISICA Y COMPUTACIONAL DE LAS INTERACCIONES PROTEINA-PROTEINA Y PROTEINA-ARNEuropean Commission http://dx.doi.org/10.13039/100011102 Seventh Framework Programme 201924 EUROPEAN DRUG INITIATIVE ON CHANNELS AND TRANSPORTERSopen accesshttp://purl.org/coar/access_right/c_abf2info:eu-repo/semantics/openAccessoai:upcommons.upc.edu:2117/1034562026-05-27T15:37:01Z
dc.title.none.fl_str_mv Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
title Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
spellingShingle Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
Rosell, Albert
Cellular & molecular mechanisms of toxin action
Amino acids--Analysis
CD98hc
4F2hc ectodomain
Heteromeric amino acid transporters (HATs)
Aminoàcids--Anàlisi
Biologia molecular
Àrees temàtiques de la UPC::Enginyeria biomèdica
title_short Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
title_full Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
title_fullStr Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
title_full_unstemmed Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
title_sort Structural bases for the interaction and stabilization of the human amino acid transporter LAT2 with its ancillary protein 4F2hc
dc.creator.none.fl_str_mv Rosell, Albert
Meury, Marcel
Alvarez-Marimon, Elena
Costa, Meritxell
Pérez-Cano, Laura
Zorzano, Antonio
Fernández-Recio, Juan
Palacín, Manuel
Fotiadis, Dimitrios
author Rosell, Albert
author_facet Rosell, Albert
Meury, Marcel
Alvarez-Marimon, Elena
Costa, Meritxell
Pérez-Cano, Laura
Zorzano, Antonio
Fernández-Recio, Juan
Palacín, Manuel
Fotiadis, Dimitrios
author_role author
author2 Meury, Marcel
Alvarez-Marimon, Elena
Costa, Meritxell
Pérez-Cano, Laura
Zorzano, Antonio
Fernández-Recio, Juan
Palacín, Manuel
Fotiadis, Dimitrios
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Cellular & molecular mechanisms of toxin action
Amino acids--Analysis
CD98hc
4F2hc ectodomain
Heteromeric amino acid transporters (HATs)
Aminoàcids--Anàlisi
Biologia molecular
Àrees temàtiques de la UPC::Enginyeria biomèdica
topic Cellular & molecular mechanisms of toxin action
Amino acids--Analysis
CD98hc
4F2hc ectodomain
Heteromeric amino acid transporters (HATs)
Aminoàcids--Anàlisi
Biologia molecular
Àrees temàtiques de la UPC::Enginyeria biomèdica
description Heteromeric amino acid transporters (HATs) are the unique example, known in all kingdoms of life, of solute transporters composed of two subunits linked by a conserved disulfide bridge. In metazoans, the heavy subunit is responsible for the trafficking of the heterodimer to the plasma membrane, and the light subunit is the transporter. HATs are involved in human pathologies such as amino acidurias, tumor growth and invasion, viral infection and cocaine addiction. However structural information about interactions between the heavy and light subunits of HATs is scarce. In this work, transmission electron microscopy and single-particle analysis of purified human 4F2hc/L-type amino acid transporter 2 (LAT2) heterodimers overexpressed in the yeast Pichia pastoris, together with docking analysis and crosslinking experiments, reveal that the extracellular domain of 4F2hc interacts with LAT2, almost completely covering the extracellular face of the transporter. 4F2hc increases the stability of the light subunit LAT2 in detergent-solubilized Pichia membranes, allowing functional reconstitution of the heterodimer into proteoliposomes. Moreover, the extracellular domain of 4F2hc suffices to stabilize solubilized LAT2. The interaction of 4F2hc with LAT2 gives insights into the structural bases for light subunit recognition and the stabilizing role of the ancillary protein in HATs.
publishDate 2014
dc.date.none.fl_str_mv 2014
2014-02-25
2017
2017-04-07
dc.type.none.fl_str_mv journal article
http://purl.org/coar/resource_type/c_6501
VoR
http://purl.org/coar/version/c_970fb48d4fbd8a85
dc.type.openaire.fl_str_mv info:eu-repo/semantics/article
format article
dc.identifier.none.fl_str_mv https://hdl.handle.net/2117/103456
https://dx.doi.org/10.1073/pnas.1323779111
url https://hdl.handle.net/2117/103456
https://dx.doi.org/10.1073/pnas.1323779111
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.relation.none.fl_str_mv Ministerio de Ciencia e Innovación http://doi.org/10.13039/501100004837 BIO2010-22324 DOCKING ENTRE PROTEINAS Y OTROS ASPECTOS: AVANCES EN LA DESCRIPCION BIOFISICA Y COMPUTACIONAL DE LAS INTERACCIONES PROTEINA-PROTEINA Y PROTEINA-ARN
European Commission http://dx.doi.org/10.13039/100011102 Seventh Framework Programme 201924 EUROPEAN DRUG INITIATIVE ON CHANNELS AND TRANSPORTERS
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
dc.rights.openaire.fl_str_mv info:eu-repo/semantics/openAccess
rights_invalid_str_mv open access
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv National Academy of Sciences
publisher.none.fl_str_mv National Academy of Sciences
dc.source.none.fl_str_mv reponame:UPCommons. Portal del coneixement obert de la UPC
instname:Universitat Politècnica de Catalunya (UPC)
instname_str Universitat Politècnica de Catalunya (UPC)
reponame_str UPCommons. Portal del coneixement obert de la UPC
collection UPCommons. Portal del coneixement obert de la UPC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15,301629

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