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
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/159784
Ver los metadatos del registro completo
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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 |
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http://sedici.unlp.edu.ar/handle/10915/159784 |
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