Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy

Autores
Vázquez, Romina Florencia; Ovalle García, Erasmo; Antillón, Armando; Ortega Blakec, Iván; Bakás, Laura Susana; Muñoz Garay, Carlos; Maté, Sabina María
Año de publicación
2021
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Sphingolipids-enriched rafts domains are proposed to occur in plasma membranes and to mediate important cellular functions. Notwithstanding, the asymmetric transbilayer distribution of phospholipids that exists in the membrane confers the two leaflets different potentials to form lateral domains as next to no sphingolipids are present in the inner leaflet. How the physical properties of one leaflet can influence the properties of the other and its importance on signal transduction across the membrane are questions still unresolved. In this work, we combined AFM imaging and Force spectroscopy measurements to assess domain formation and to study the nanomechanical properties of asymmetric supported lipid bilayers (SLBs) mimicking membrane rafts. Asymmetric SLBs were formed by incorporating N-palmitoyl-sphingomyelin (16:0SM) into the outer leaflet of preformed 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/Cholesterol SLBs through methyl-β-cyclodextrin– mediated lipid exchange. Lipid domains were detected after incorporation of 16:0SM though their phase state varied from gel to liquid ordered (Lo) phase if the procedure was performed at 24 or 37 °C, respectively. When comparing symmetric and asymmetric Lo domains, differences in size and morphology were observed, with asymmetric domains being smaller and more interconnected. Both types of Lo domains showed similar mechanical stability in terms of rupture forces and Young's moduli. Notably, force curves in asymmetric domains presented two rupture events that could be attributed to the sequential rupture of a liquid disordered (Ld) and a Lo phase. Interleaflet coupling in asymmetric Lo domains could also be inferred from those measurements. The experimental approach outlined here would significantly enhance the applicability of membrane models.
Instituto de Investigaciones Bioquímicas de La Plata
Centro de Investigación de Proteínas Vegetales
Materia
Bioquímica
Lipid asymmetry
Sphingomyelin
Lipid domains
Supported lipid bilayers
Atomic force microscopy
Force spectroscopy
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/159784

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network_name_str SEDICI (UNLP)
spelling Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopyVázquez, Romina FlorenciaOvalle García, ErasmoAntillón, ArmandoOrtega Blakec, IvánBakás, Laura SusanaMuñoz Garay, CarlosMaté, Sabina MaríaBioquímicaLipid asymmetrySphingomyelinLipid domainsSupported lipid bilayersAtomic force microscopyForce spectroscopySphingolipids-enriched rafts domains are proposed to occur in plasma membranes and to mediate important cellular functions. Notwithstanding, the asymmetric transbilayer distribution of phospholipids that exists in the membrane confers the two leaflets different potentials to form lateral domains as next to no sphingolipids are present in the inner leaflet. How the physical properties of one leaflet can influence the properties of the other and its importance on signal transduction across the membrane are questions still unresolved. In this work, we combined AFM imaging and Force spectroscopy measurements to assess domain formation and to study the nanomechanical properties of asymmetric supported lipid bilayers (SLBs) mimicking membrane rafts. Asymmetric SLBs were formed by incorporating N-palmitoyl-sphingomyelin (16:0SM) into the outer leaflet of preformed 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/Cholesterol SLBs through methyl-β-cyclodextrin– mediated lipid exchange. Lipid domains were detected after incorporation of 16:0SM though their phase state varied from gel to liquid ordered (Lo) phase if the procedure was performed at 24 or 37 °C, respectively. When comparing symmetric and asymmetric Lo domains, differences in size and morphology were observed, with asymmetric domains being smaller and more interconnected. Both types of Lo domains showed similar mechanical stability in terms of rupture forces and Young's moduli. Notably, force curves in asymmetric domains presented two rupture events that could be attributed to the sequential rupture of a liquid disordered (Ld) and a Lo phase. Interleaflet coupling in asymmetric Lo domains could also be inferred from those measurements. The experimental approach outlined here would significantly enhance the applicability of membrane models.Instituto de Investigaciones Bioquímicas de La PlataCentro de Investigación de Proteínas Vegetales2021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://sedici.unlp.edu.ar/handle/10915/159784enginfo:eu-repo/semantics/altIdentifier/issn/0005-2736info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2020.183467info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:41:46Zoai:sedici.unlp.edu.ar:10915/159784Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:41:46.817SEDICI (UNLP) - Universidad Nacional de La Platafalse
dc.title.none.fl_str_mv Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
title Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
spellingShingle Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
Vázquez, Romina Florencia
Bioquímica
Lipid asymmetry
Sphingomyelin
Lipid domains
Supported lipid bilayers
Atomic force microscopy
Force spectroscopy
title_short Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
title_full Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
title_fullStr Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
title_full_unstemmed Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
title_sort Asymmetric bilayers mimicking membrane rafts prepared by lipid exchange: nanoscale characterization using AFM-Force spectroscopy
dc.creator.none.fl_str_mv Vázquez, Romina Florencia
Ovalle García, Erasmo
Antillón, Armando
Ortega Blakec, Iván
Bakás, Laura Susana
Muñoz Garay, Carlos
Maté, Sabina María
author Vázquez, Romina Florencia
author_facet Vázquez, Romina Florencia
Ovalle García, Erasmo
Antillón, Armando
Ortega Blakec, Iván
Bakás, Laura Susana
Muñoz Garay, Carlos
Maté, Sabina María
author_role author
author2 Ovalle García, Erasmo
Antillón, Armando
Ortega Blakec, Iván
Bakás, Laura Susana
Muñoz Garay, Carlos
Maté, Sabina María
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Bioquímica
Lipid asymmetry
Sphingomyelin
Lipid domains
Supported lipid bilayers
Atomic force microscopy
Force spectroscopy
topic Bioquímica
Lipid asymmetry
Sphingomyelin
Lipid domains
Supported lipid bilayers
Atomic force microscopy
Force spectroscopy
dc.description.none.fl_txt_mv Sphingolipids-enriched rafts domains are proposed to occur in plasma membranes and to mediate important cellular functions. Notwithstanding, the asymmetric transbilayer distribution of phospholipids that exists in the membrane confers the two leaflets different potentials to form lateral domains as next to no sphingolipids are present in the inner leaflet. How the physical properties of one leaflet can influence the properties of the other and its importance on signal transduction across the membrane are questions still unresolved. In this work, we combined AFM imaging and Force spectroscopy measurements to assess domain formation and to study the nanomechanical properties of asymmetric supported lipid bilayers (SLBs) mimicking membrane rafts. Asymmetric SLBs were formed by incorporating N-palmitoyl-sphingomyelin (16:0SM) into the outer leaflet of preformed 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/Cholesterol SLBs through methyl-β-cyclodextrin– mediated lipid exchange. Lipid domains were detected after incorporation of 16:0SM though their phase state varied from gel to liquid ordered (Lo) phase if the procedure was performed at 24 or 37 °C, respectively. When comparing symmetric and asymmetric Lo domains, differences in size and morphology were observed, with asymmetric domains being smaller and more interconnected. Both types of Lo domains showed similar mechanical stability in terms of rupture forces and Young's moduli. Notably, force curves in asymmetric domains presented two rupture events that could be attributed to the sequential rupture of a liquid disordered (Ld) and a Lo phase. Interleaflet coupling in asymmetric Lo domains could also be inferred from those measurements. The experimental approach outlined here would significantly enhance the applicability of membrane models.
Instituto de Investigaciones Bioquímicas de La Plata
Centro de Investigación de Proteínas Vegetales
description Sphingolipids-enriched rafts domains are proposed to occur in plasma membranes and to mediate important cellular functions. Notwithstanding, the asymmetric transbilayer distribution of phospholipids that exists in the membrane confers the two leaflets different potentials to form lateral domains as next to no sphingolipids are present in the inner leaflet. How the physical properties of one leaflet can influence the properties of the other and its importance on signal transduction across the membrane are questions still unresolved. In this work, we combined AFM imaging and Force spectroscopy measurements to assess domain formation and to study the nanomechanical properties of asymmetric supported lipid bilayers (SLBs) mimicking membrane rafts. Asymmetric SLBs were formed by incorporating N-palmitoyl-sphingomyelin (16:0SM) into the outer leaflet of preformed 1,2-Dioleoyl-sn-glycero-3-phosphocholine (DOPC)/Cholesterol SLBs through methyl-β-cyclodextrin– mediated lipid exchange. Lipid domains were detected after incorporation of 16:0SM though their phase state varied from gel to liquid ordered (Lo) phase if the procedure was performed at 24 or 37 °C, respectively. When comparing symmetric and asymmetric Lo domains, differences in size and morphology were observed, with asymmetric domains being smaller and more interconnected. Both types of Lo domains showed similar mechanical stability in terms of rupture forces and Young's moduli. Notably, force curves in asymmetric domains presented two rupture events that could be attributed to the sequential rupture of a liquid disordered (Ld) and a Lo phase. Interleaflet coupling in asymmetric Lo domains could also be inferred from those measurements. The experimental approach outlined here would significantly enhance the applicability of membrane models.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
Articulo
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://sedici.unlp.edu.ar/handle/10915/159784
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dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/issn/0005-2736
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bbamem.2020.183467
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
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rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
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