Brij detergents reveal new aspects of membrane microdomain in erythrocytes

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dc.contributor.authorCasadei, Bruna Renata
dc.contributor.authorCarvalho, Patricia de Oliveira
dc.contributor.authorRiske, Karin A. [UNIFESP]
dc.contributor.authorBarbosa, Raquel de Melo
dc.contributor.authorDe Paula, Eneida
dc.contributor.authorDomingues, Cleyton Crepaldi
dc.contributor.institutionUniversidade Estadual de Campinas (UNICAMP)
dc.contributor.institutionUniv Sao Francisco
dc.contributor.institutionUniversidade Federal de São Paulo (UNIFESP)
dc.date.accessioned2016-01-24T14:37:51Z
dc.date.available2016-01-24T14:37:51Z
dc.date.issued2014-09-01
dc.description.abstractMembrane microdomains enriched in cholesterol, sphingolipids (rafts), and specific proteins are involved in important physiological functions. However their structure, size and stability are still controversial. Given that detergent-esistant membranes (DRMs) are in the liquid-ordered state and are rich in raft-like components, they might correspond to rafts at least to some extent. Here we monitor the lateral order of biological membranes by characterizing DRMs from erythrocytes obtained with Brij-98, Brij-58, and TX-100 at 4 degrees C and 37 degrees C. All DRMs were enriched in cholesterol and contained the raft markers flotillin-2 and stomatin. However, sphingomyelin (SM) was only found to be enriched in TX-100-DRMs -a detergent that preferentially solubilizes the membrane inner leaflet -while Band 3 was present solely in Brij-DRMs. Electron paramagnetic resonance spectra showed that the acyl chain packing of Brij-DRMs was lower than TX-100-DRMs, providing evidence of their diverse lipid composition. Fatty acid analysis revealed that the SM fraction of the DRMs was enriched in lignoceric acid, which should specifically contribute to the resistance of SM to detergents. These results indicate that lipids from the outer leaflet, particularly SM, are essential for the formation of the liquid-ordered phase of DRMs. At last, the differential solubilization process induced by Brij-98 and TX-100 was monitored using giant unilamellar vesicles. This study suggests that Brij and TX-100-DRMs reflect different degrees of lateral order of the membrane microdomains. Additionally, Brij DRMs are composed by both inner and outer leaflet components, making them more physiologically relevant than TX-100-DRMs to the studies of membrane rafts.en
dc.description.affiliationUniv Estadual Campinas Unicamp, Inst Biol, Dept Bioquim & Biol Tecidual, Campinas, SP, Brazil
dc.description.affiliationUniv Sao Francisco, Lab Multidisciplinar Pesquisa, BR-12900000 Braganca Paulista, SP, Brazil
dc.description.affiliationUniversidade Federal de São Paulo Unifesp, Dept Biofis, São Paulo, Brazil
dc.description.affiliationUnifespUniversidade Federal de São Paulo Unifesp, Dept Biofis, São Paulo, Brazil
dc.description.sourceWeb of Science
dc.description.sponsorshipFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorshipConselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorshipIDFAPESP: 2009/00904-1
dc.description.sponsorshipIDFAPESP: 2010/18516-5
dc.description.sponsorshipIDCNPq: 479993/2011-4
dc.description.sponsorshipIDCNPq: 132836/2009-2
dc.format.extent195-205
dc.identifierhttp://dx.doi.org/10.3109/09687688.2014.949319
dc.identifier.citationMolecular Membrane Biology. London: Informa Healthcare, v. 31, n. 6, p. 195-205, 2014.
dc.identifier.doi10.3109/09687688.2014.949319
dc.identifier.issn0968-7688
dc.identifier.urihttp://repositorio.unifesp.br/handle/11600/38204
dc.identifier.wosWOS:000342203800003
dc.language.isoeng
dc.publisherInforma Healthcare
dc.relation.ispartofMolecular Membrane Biology
dc.rightsinfo:eu-repo/semantics/restrictedAccess
dc.rights.licensehttp://informahealthcare.com/userimages/ContentEditor/1255620309227/Copyright_And_Permissions.pdf
dc.subjectDetergenten
dc.subjecterythrocyteen
dc.subjectlipid raftsen
dc.subjectlipid bilayeren
dc.titleBrij detergents reveal new aspects of membrane microdomain in erythrocytesen
dc.typeinfo:eu-repo/semantics/article
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