Lipid Bilayers in the Gel Phase Become Saturated by Triton X-100 at Lower Surfactant Concentrations Than Those in the Fluid Phase

Ahyayauch, Hasna; Collado, M. Isabel; Alonso, Alicia; Goñi, Félix M.

doi:10.1016/j.bpj.2012.04.041

Cited by 29 publications

(31 citation statements)

References 39 publications

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“…The results show that POPC, POPC/chol and SM become completely permeable (0% fluorescence intensity) below the onset of solubilization, which is detected when sample light scattering starts to decrease. POPC and SM are completely solubilized by TX-100, although more TX-100 is required to fully solubilize the Ld phase, as reported previously (Ahyayauch et al 2012). On the other hand, the solubilization of POPC/chol is not complete, as seen from the residual light scattering value (~20%) at high detergent concentration.…”

Section: Solubilization Of Binary Lipid Mixtures Containing Cholesterolsupporting

confidence: 78%

“…Among the detergents studied, TX-100 has attracted special attention because of its ubiquitous use in solubilization protocols (Hjelmeland 1990). The solubilization of membranes by TX-100 has been shown to be highly dependent on the lipid bilayer composition (Sot et al 2002;Heerklotz 2002;Heerklotz et al 2003;Tsamaloukas et al 2006;Arnulphi et al 2007;Ahyayauch et al 2012). Especially, the presence of cholesterol has a great impact on the membrane resistance to effective and complete solubilization (Patra et al 1999;Li et al 2001;Schnitzer et al 2005;El Kirat and Morandat 2007;Ahyayauch et al 2009).…”

Section: Drms-associated Proteinsmentioning

confidence: 99%

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Biophysical approaches in the study of biomembrane solubilization: quantitative assessment and the role of lateral inhomogeneity

et al. 2017

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Detergents are amphiphilic molecules widely used to solubilize biological membranes and/or extract their components. Nevertheless, because of the complex composition of biomembranes, their solubilization by detergents has not been systematically studied. In this review, we address the solubilization of erythrocytes, which provide a relatively simple, robust and easy to handle biomembrane, and of biomimetic models, to stress the role of the lipid composition on the solubilization process. First, results of a systematic study on the solubilization of human erythrocyte membranes by different series of non-ionic (Triton, CxEy, Brij, Renex, Tween), anionic (bile salts) and zwitterionic (ASB, CHAPS) detergents are shown. Such quantitative approach allowed us to propose R e sat -the effective detergent/lipid molar ratio in the membrane for the onset of hemolysis as a new parameter to classify the solubilization efficiency of detergents. Second, detergent-resistant membranes (DRMs) obtained as a result of the partial solubilization of erythrocytes by TX-100, C 12 E 8 and Brij detergents are examined. DRMs were characterized by their cholesterol, sphingolipid and specific proteins content, as well as lipid packing. Finally, lipid bilayers of tuned lipid composition forming liposomes were used to investigate the solubilization process of membranes of different compositions/phases induced by Triton X-100. Optical microscopy of giant unilamellar vesicles revealed that pure phospholipid membranes are fully solubilized, whereas the presence of cholesterol renders the mixture partially or even fully insoluble, depending on the composition. Additionally, Triton X-100 induced phase separation in raft-like mixtures, and selective solubilization of the fluid phase only.

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Section: Solubilization Of Binary Lipid Mixtures Containing Cholesterolsupporting

confidence: 78%

Section: Drms-associated Proteinsmentioning

confidence: 99%

Biophysical approaches in the study of biomembrane solubilization: quantitative assessment and the role of lateral inhomogeneity

et al. 2017

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“…It is unfortunate that the experimental data in this region could not be fit to any simple partition model, thus could not be quantitatively interpreted. Moreover, lipid bilayers in the gel phase become saturated by Triton X-100 at lower surfactant concentrations than those in the fluid phase (36). Again, this remains clearly an area open to further research.…”

Section: Lipid Melting Points and Detergent Solubilizationmentioning

confidence: 98%

High-Melting Lipid Mixtures and the Origin of Detergent-Resistant Membranes Studied with Temperature-Solubilization Diagrams

Sot

Manni

Viguera

et al. 2014

Biophysical Journal

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The origin of resistance to detergent solubilization in certain membranes, or membrane components, is not clearly understood. We have studied the solubilization by Triton X-100 of binary mixtures composed of egg sphingomyelin (SM) and either ceramide, diacylglycerol, or cholesterol. Solubilization has been assayed in the 4-50°C range, and the results are summarized in a novel, to our knowledge, form of plots, that we have called temperature-solubilization diagrams. Despite using a large detergent excess (lipid/detergent 1:20 mol ratio) and extended solubilization times (24-48 h) certain mixtures were not amenable to Triton X-100 solubilization at one or more temperatures. DSC of all the lipid mixtures, and of all the lipid + detergent mixtures revealed that detergent resistance was associated with the presence of gel domains at the assay temperature. Once the system melted down, solubilization could occur. In general adding high-melting lipids limited the solubilization, whereas the addition of low-melting lipids promoted it. Lipidomic analysis of Madin-Darby canine kidney cell membranes and of the corresponding detergent-resistant fraction indicated a large enrichment of the nonsolubilized components in saturated diacylglycerol and ceramide. SM-cholesterol mixtures were special in that detergent solubilization was accompanied, for certain temperatures and compositions, by an independent phenomenon of reassembly of the partially solubilized lipid bilayers. The temperature at which lysis and reassembly prevailed was ∼25°C, thus for some SM-cholesterol mixtures solubilization occurred both above and below 25°C, but not at that temperature. These observations can be at the origin of the detergent resistance effects observed with cell membranes, and they also mean that cholesterol-containing detergent-resistant membrane remnants cannot correspond to structures existing in the native membrane before detergent addition.

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“…This product contains deionized water, citrate buffer salts, and diethanolamine, a compound that is used as a surfactant. 5,8,10 EnVision TM FLEX Target Retrieval Solution, Low pH is an all-in-one epitope retrieval buffer that served as a control for surfactant performance. HIER L buffer contains deionized water and citrate buffer salts.…”

Section: Paraffin Solubilitymentioning

confidence: 99%

“…Carbowax TM , polyethylene glycol) do reduce the surface tension in the buffers and help emulsify molten paraffin. 3,4,[7][8][9] But this is not enough activity to dissolve pooled molten paraffin. Specifically, paraffins that contain higher amounts of plastic polymers have been found to be particularly difficult to remove using the traditional all-in-one buffer formulations.…”

Section: Introductionmentioning

confidence: 96%

Water-soluble organic solvent improves paraffin displacement in all-in-one epitope retrieval buffers

Freeland¹,

Bueren²,

Shanmugakonar

et al. 2013

Journal of Histotechnology

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An all-in-one epitope retrieval buffer was developed to solubilize molten paraffin. This study illustrates that common all-in-one buffer additives such as surfactants and wetting agents do not solubilize molten paraffin but a water-soluble organic solvent (WSOS) does. This study also shows that the WSOS does not negatively impact immunohistochemical staining reactions. The addition of a WSOS into an all-in-one epitope retrieval buffer formulation maintains paraffin in solution when slides mounted with tissue sections are removed from the buffer. This action is an advantage for buffer tanks in which slides are in a vertical orientation; the slides are not recoated with molten paraffin.

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Lipid Bilayers in the Gel Phase Become Saturated by Triton X-100 at Lower Surfactant Concentrations Than Those in the Fluid Phase

Cited by 29 publications

References 39 publications

Biophysical approaches in the study of biomembrane solubilization: quantitative assessment and the role of lateral inhomogeneity

Biophysical approaches in the study of biomembrane solubilization: quantitative assessment and the role of lateral inhomogeneity

High-Melting Lipid Mixtures and the Origin of Detergent-Resistant Membranes Studied with Temperature-Solubilization Diagrams

Water-soluble organic solvent improves paraffin displacement in all-in-one epitope retrieval buffers

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