Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles

Considerable attention has been dedicated to lipid rafts due to their importance in numerous cell functions such as membrane trafficking, polarization, and signaling. Next to studies in living cells, artificial micrometer-sized vesicles with a minimal set of components are established as a major too...

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Autores: Otrin, Nika, Bednarz, Claudia, Otrin, Lado, Ivanov, Ivan|||0000-0002-4675-5287, Träger, Toni K., Hamdi, Farzad, Kastritis, Panagiotis L., Sundmacher, Kai
Formato: artículo
Fecha de publicación:2024
País:España
Recursos:Universitat Politècnica de Catalunya (UPC)
Repositorio:UPCommons. Portal del coneixement obert de la UPC
Idioma:inglés
OAI Identifier:oai:upcommons.upc.edu:2117/403768
Acesso em linha:https://hdl.handle.net/2117/403768
https://dx.doi.org/10.1021/acs.biomac.3c00972
Access Level:acceso abierto
Palavra-chave:Peptides
Lipids
Proteins
Membranes
Peptides and proteins
Phase separation
Vesicles
Pèptids
Lípids
Proteïnes
Àrees temàtiques de la UPC::Enginyeria química
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repository_id_str
spelling Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesiclesOtrin, NikaBednarz, ClaudiaOtrin, LadoIvanov, Ivan|||0000-0002-4675-5287Träger, Toni K.Hamdi, FarzadKastritis, Panagiotis L.Sundmacher, KaiPeptidesLipidsProteinsLipidsMembranesPeptides and proteinsPhase separationVesiclesPèptidsLípidsProteïnesÀrees temàtiques de la UPC::Enginyeria químicaConsiderable attention has been dedicated to lipid rafts due to their importance in numerous cell functions such as membrane trafficking, polarization, and signaling. Next to studies in living cells, artificial micrometer-sized vesicles with a minimal set of components are established as a major tool to understand the phase separation dynamics and their intimate interplay with membrane proteins. In parallel, mixtures of phospholipids and certain amphiphilic polymers simultaneously offer an interface for proteins and mimic this segregation behavior, presenting a tangible synthetic alternative for fundamental studies and bottom-up design of cellular mimics. However, the simultaneous insertion of complex and sensitive membrane proteins is experimentally challenging and thus far has been largely limited to natural lipids. Here, we present the co-reconstitution of the proton pump bo3 oxidase and the proton consumer ATP synthase in hybrid polymer/lipid giant unilamellar vesicles (GUVs) via fusion/electroformation. Variations of the current method allow for tailored reconstitution protocols and control of the vesicle size. In particular, mixing of protein-free and protein-functionalized nanosized vesicles in the electroformation film results in larger GUVs, while separate reconstitution of the respiratory enzymes enables higher ATP synthesis rates. Furthermore, protein labeling provides a synthetic mechanism for phase separation and protein sequestration, mimicking lipid- and protein-mediated domain formation in nature. The latter means opens further possibilities for re-enacting phenomena like supercomplex assembly or symmetry breaking and enriches the toolbox of bottom-up synthetic biology.This work is funded by the Federal Ministry of Education and Research (BMBF) of Germany and the Max Planck Society. K.S. acknowledges funding from the Max Planck School Matter to Life, a joint graduate program of German Universities and Research Organizations. This work was supported by the European Union through funding from the Horizon Europe ERA Chair “hot4cryo” project number 101086665 (to P.L.K.), the Federal Ministry of Education and Research (BMBF, ZIK program) (Grant nos. 03Z22HN23, 03Z22HI2, and 03COV04 to P.L.K.), the European Regional Development Funds (EFRE) for Saxony-Anhalt (Grant no. ZS/2016/04/78115 to P.L.K.), the Deutsche Forschungsgemeinschaft (project number 391498659, RTG 2467), and the Martin-Luther University of Halle-Wittenberg. The authors are grateful to Anne Christin Reichelt for her help with microscopy and Dr. Christian Tüting for his help with cryo-TEM.Peer Reviewed20242024-01-0820242024-03-06journal articlehttp://purl.org/coar/resource_type/c_6501VoRhttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/2117/403768https://dx.doi.org/10.1021/acs.biomac.3c00972reponame:UPCommons. Portal del coneixement obert de la UPCinstname:Universitat Politècnica de Catalunya (UPC)Inglésengopen accesshttp://purl.org/coar/access_right/c_abf2Attribution 4.0 Internationalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:upcommons.upc.edu:2117/4037682026-05-27T15:37:01Z
dc.title.none.fl_str_mv Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
title Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
spellingShingle Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
Otrin, Nika
Peptides
Lipids
Proteins
Lipids
Membranes
Peptides and proteins
Phase separation
Vesicles
Pèptids
Lípids
Proteïnes
Àrees temàtiques de la UPC::Enginyeria química
title_short Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
title_full Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
title_fullStr Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
title_full_unstemmed Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
title_sort Protein-rich rafts in hybrid polymer/lipid giant unilamellar vesicles
dc.creator.none.fl_str_mv Otrin, Nika
Bednarz, Claudia
Otrin, Lado
Ivanov, Ivan|||0000-0002-4675-5287
Träger, Toni K.
Hamdi, Farzad
Kastritis, Panagiotis L.
Sundmacher, Kai
author Otrin, Nika
author_facet Otrin, Nika
Bednarz, Claudia
Otrin, Lado
Ivanov, Ivan|||0000-0002-4675-5287
Träger, Toni K.
Hamdi, Farzad
Kastritis, Panagiotis L.
Sundmacher, Kai
author_role author
author2 Bednarz, Claudia
Otrin, Lado
Ivanov, Ivan|||0000-0002-4675-5287
Träger, Toni K.
Hamdi, Farzad
Kastritis, Panagiotis L.
Sundmacher, Kai
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Peptides
Lipids
Proteins
Lipids
Membranes
Peptides and proteins
Phase separation
Vesicles
Pèptids
Lípids
Proteïnes
Àrees temàtiques de la UPC::Enginyeria química
topic Peptides
Lipids
Proteins
Lipids
Membranes
Peptides and proteins
Phase separation
Vesicles
Pèptids
Lípids
Proteïnes
Àrees temàtiques de la UPC::Enginyeria química
description Considerable attention has been dedicated to lipid rafts due to their importance in numerous cell functions such as membrane trafficking, polarization, and signaling. Next to studies in living cells, artificial micrometer-sized vesicles with a minimal set of components are established as a major tool to understand the phase separation dynamics and their intimate interplay with membrane proteins. In parallel, mixtures of phospholipids and certain amphiphilic polymers simultaneously offer an interface for proteins and mimic this segregation behavior, presenting a tangible synthetic alternative for fundamental studies and bottom-up design of cellular mimics. However, the simultaneous insertion of complex and sensitive membrane proteins is experimentally challenging and thus far has been largely limited to natural lipids. Here, we present the co-reconstitution of the proton pump bo3 oxidase and the proton consumer ATP synthase in hybrid polymer/lipid giant unilamellar vesicles (GUVs) via fusion/electroformation. Variations of the current method allow for tailored reconstitution protocols and control of the vesicle size. In particular, mixing of protein-free and protein-functionalized nanosized vesicles in the electroformation film results in larger GUVs, while separate reconstitution of the respiratory enzymes enables higher ATP synthesis rates. Furthermore, protein labeling provides a synthetic mechanism for phase separation and protein sequestration, mimicking lipid- and protein-mediated domain formation in nature. The latter means opens further possibilities for re-enacting phenomena like supercomplex assembly or symmetry breaking and enriches the toolbox of bottom-up synthetic biology.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024-01-08
2024
2024-03-06
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/2117/403768
https://dx.doi.org/10.1021/acs.biomac.3c00972
url https://hdl.handle.net/2117/403768
https://dx.doi.org/10.1021/acs.biomac.3c00972
dc.language.none.fl_str_mv Inglés
eng
language_invalid_str_mv Inglés
language eng
dc.rights.none.fl_str_mv open access
http://purl.org/coar/access_right/c_abf2
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
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
Attribution 4.0 International
http://creativecommons.org/licenses/by/4.0/
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
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 15,301603

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