Direct Visualization of the Action of Triton X-100 on Giant Vesicles of Erythrocyte Membrane Lipids

Show simple item record Casadei, Bruna R. Domingues, Cleyton C. Paula, Eneida de Riske, Karin do Amaral [UNIFESP] 2016-01-24T14:37:24Z 2016-01-24T14:37:24Z 2014-06-03
dc.identifier.citation Biophysical Journal. Cambridge: Cell Press, v. 106, n. 11, p. 2417-2425, 2014.
dc.identifier.issn 0006-3495
dc.description.abstract The raft hypothesis proposes that microdomains enriched in sphingolipids, cholesterol, and specific proteins are transiently formed to accomplish important cellular tasks. Equivocally, detergent-resistant membranes were initially assumed to be identical to membrane rafts, because of similarities between their compositions. in fact, the impact of detergents in membrane organization is still controversial. Here, we use phase contrast and fluorescence microscopy to observe giant unilamellar vesicles (GUVs) made of erythrocyte membrane lipids (erythro-GUVs) when exposed to the detergent Triton X-100 (TX-100). We clearly show that TX-100 has a restructuring action on biomembranes. Contact with TX-100 readily induces domain formation on the previously homogeneous membrane of erythro-GUVs at physiological and room temperatures. the shape and dynamics of the formed domains point to liquid-ordered/liquid-disordered (Lo/Ld) phase separation, typically found in raft-like ternary lipid mixtures. the Ld domains are then separated from the original vesicle and completely solubilized by TX-100. the insoluble vesicle left, in the Lo phase, represents around 2/3 of the original vesicle surface at room temperature and decreases to almost 1/2 at physiological temperature. This chain of events could be entirely reproduced with biomimetic GUVs of a simple ternary lipid mixture, 2:1:2 POPC/SM/chol (phosphatidylcholine/sphyngonnyelin/cholesterol), showing that this behavior will arise because of fundamental physicochemical properties of simple lipid mixtures. This work provides direct visualization of TX-100-induced domain formation followed by selective (Ld phase) solubilization in a model system with a complex biological lipid composition. en
dc.description.sponsorship Brazilian Research foundations
dc.description.sponsorship Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
dc.description.sponsorship Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
dc.description.sponsorship TNCT-Fax
dc.description.sponsorship Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
dc.format.extent 2417-2425
dc.language.iso eng
dc.publisher Cell Press
dc.relation.ispartof Biophysical Journal
dc.rights Acesso aberto
dc.title Direct Visualization of the Action of Triton X-100 on Giant Vesicles of Erythrocyte Membrane Lipids en
dc.type Artigo
dc.contributor.institution Universidade Estadual de Campinas (UNICAMP)
dc.contributor.institution Universidade Federal de São Paulo (UNIFESP)
dc.description.affiliation Univ Estadual Campinas, Inst Biol, Dept Bioquim, Campinas, SP, Brazil
dc.description.affiliation Universidade Federal de São Paulo, Dept Biofis, São Paulo, Brazil
dc.description.affiliationUnifesp Universidade Federal de São Paulo, Dept Biofis, São Paulo, Brazil
dc.description.sponsorshipID FAPESP: 2009/0904-1
dc.description.sponsorshipID FAPESP: 2012/10442-8
dc.description.sponsorshipID FAPESP: 2010/18516-5
dc.description.sponsorshipID CNPq: 479993/2011-4
dc.description.sponsorshipID CNPq: 472054/2011-2
dc.identifier.file WOS000337012300015.pdf
dc.identifier.doi 10.1016/j.bpj.2014.04.039
dc.description.source Web of Science
dc.identifier.wos WOS:000337012300015


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