Solubilization of binary lipid mixtures by the detergent Triton X-100 : the role of cholesterol

The solubilization of lipid bilayers of different composition and phase by the detergent Triton X-100 (Triton X-100) was investigated using optical and fluorescence microscopy of giant unilamellar vesicles (GUVs) and light scattering of large unilamellar vesicles (LUVs). The compositions explored we...

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Publié dans:Langmuir : the ACS journal of surfaces and colloids. - 1985. - 31(2015), 1 vom: 07., Seite 378-86
Auteur principal: Mattei, Bruno (Auteur)
Autres auteurs: França, Ana D C, Riske, Karin A
Format: Article en ligne
Langue:English
Publié: 2015
Accès à la collection:Langmuir : the ACS journal of surfaces and colloids
Sujets:Journal Article Research Support, Non-U.S. Gov't Lipids Octoxynol 9002-93-1 Cholesterol 97C5T2UQ7J
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520 |a The solubilization of lipid bilayers of different composition and phase by the detergent Triton X-100 (Triton X-100) was investigated using optical and fluorescence microscopy of giant unilamellar vesicles (GUVs) and light scattering of large unilamellar vesicles (LUVs). The compositions explored were 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), in the liquid-disordered (Ld) phase, sphingomyelin (SM), in the gel phase, and binary mixtures of these phospholipids with 30 mol % cholesterol (chol), resulting in bilayers in the Ld and liquid-ordered (Lo) phases, respectively. We show that the phospholipid bilayers are completely soluble in TX-100, but optical microscopy reveals that whereas fluid POPC is gradually solubilized by TX-100, gel SM is first shattered in bilayer fragments. Incorporation of TX-100 in the membrane leads to increase in GUV area, which was quantified and expressed as bound detergent-to-lipid molar ratio. The partition of TX-100 in POPC is very high, decreases in POPC/chol, and is negligible in SM/chol. Fluorescence microscopy shows that TX-100 induces Lo/Ld phase separation in previously homogeneous POPC/chol GUVs, and insoluble bilayer fragments/vesicles are detected with optical microscopy and light scattering. Vesicles of SM/chol, in the Lo phase, are virtually insoluble in TX-100. Taken together, our results show that the presence of cholesterol is the origin of membrane resistance to solubilization, which depending on the specific lipid mixture can result in either partially (POPC/chol) or completely (SM/chol) insoluble mixtures 
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