A universal method for planar lipid bilayer formation by freeze and thaw

Sugihara, Kaori; Jang, Bumjin; Schneider, Manuel; Vörös, János; Zambelli, Tomaso

doi:10.1039/c2sm25148e

Cited by 22 publications

(31 citation statements)

References 52 publications

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“…4 a). This case is analogous to the formation of an intact vesicle layer whereby adsorbed vesicles do not rupture and, as a result, lipid molecules within the adlayer are immobile and there is also no fluorescence recovery 35 , 52 , 53 . In marked contrast, the fluorescence intensity within the bleached spot was quickly recovered for SLB adlayers formed using bicelles at q = 0.25 (Fig.…”

Section: Resultsmentioning

confidence: 99%

Versatile formation of supported lipid bilayers from bicellar mixtures of phospholipids and capric acid

Sut

Yoon

Park

et al. 2020

Sci Rep

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Originally developed for the structural biology field, lipid bicelle nanostructures composed of longand short-chain phospholipid molecules have emerged as a useful interfacial science tool to fabricate two-dimensional supported lipid bilayers (SLBs) on hydrophilic surfaces due to ease of sample preparation, scalability, and versatility. To improve SLB fabrication prospects, there has been recent interest in replacing the synthetic, short-chain phospholipid component of bicellar mixtures with naturally abundant fatty acids and monoglycerides, i.e., lauric acid and monocaprin. Such options have proven successful under specific conditions, however, there is room for devising more versatile fabrication options, especially in terms of overcoming lipid concentration-dependent SLB formation limitations. Herein, we investigated SLB fabrication by using bicellar mixtures consisting of long-chain phospholipid and capric acid, the latter of which has similar headgroup and chain length properties to lauric acid and monocaprin, respectively. Quartz crystal microbalance-dissipation, epifluorescence microscopy, and fluorescence recovery after photobleaching experiments were conducted to characterize lipid concentration-dependent bicelle adsorption onto silicon dioxide surfaces. We identified that uniform-phase SLB formation occurred independently of total lipid concentration when the ratio of long-chain phospholipid to capric acid molecules ("q-ratio") was 0.25 or 2.5, which is superior to past results with lauric acid-and monocaprin-containing bicelles in which cases lipid concentration-dependent behavior was observed. Together, these findings demonstrate that capric acid-containing bicelles are versatile tools for SLB fabrication and highlight how the molecular structure of bicelle components can be rationally finetuned to modulate self-assembly processes at solid-liquid interfaces. Bicelles are an important class of membrane-mimicking lipid nanostructures that self-assemble from mixtures of long-and short-chain phospholipids under appropriate processing conditions 1,2. Also known as lipid nanodisks 3 or bilayered mixed micelles 4,5 , bicelles can exist in a wide range of morphologies (e.g., perforated sheets and wormlike micelles) depending on parameters such as temperature, q-ratio (long-to short-chain phospholipid molar ratio), total lipid concentration, and lipid composition, and are widely conceptualized as two-dimensional disks whereby long-chain phospholipids constitute a planar lipid bilayer surface and the short-chain phospholipids form a rimmed edge around the bilayer 6-12. Since they can exhibit magnetic alignment in some cases, bicellar disks have long been used in the nuclear magnetic resonance spectroscopy field as membrane protein hosts 4,13-16. In the interfacial science field, bicelles have also proven to be useful as a lipid nanostructure to fabricate supported lipid bilayers (SLBs), which are extensively used in applications such as biosensors and micropatterned arrays 17-25. Indeed, bicelle adsorption onto...

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Section: Resultsmentioning

confidence: 99%

Versatile formation of supported lipid bilayers from bicellar mixtures of phospholipids and capric acid

Sut

Yoon

Park

et al. 2020

Sci Rep

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“…The third process of the adsorbed vesicles on substrates is an adhesive vesicular layer that stably forms, and the transformation to a planar membrane does not proceed ( Figure 1 c) [ 59 , 63 ]. Once the vesicular layer forms, it rarely turns to a planar membrane spontaneously, but there are several known ways to stimulate adhesive vesicles to rupture: addition of a reagent working as a membrane fuser (Ca 2+ [ 58 ], poly-ethylene glycol [ 68 ]) or an amphipathic viral peptide [ 69 ], osmotic pressure [ 60 ], mild sonication [ 70 ] and freeze-and-thaw [ 71 ].…”

Section: Substrate Effects On Slb Formation Processmentioning

confidence: 99%

“…Fluid SLBs forms only when the density of charged termination (–NH 2 or –COOH) is ≥ 80% mixed with the neutral termination (–OH). The surface of TiO 2 is also hydrophilic, but previous studies from several groups showed that the adhesive vesicular layer of PC is formed predominantly on sputter-deposited TiO 2 surfaces [ 40 , 59 , 60 , 69 , 71 , 82 , 83 ]. Rossetti et al reported that a SLB is formed from vesicles on a sputter-deposited TiO 2 when the vesicle contains 20% of negatively charged lipid, phosphatidylserine (PS), in the presence of Ca 2+ ion in the buffer solution [ 40 ].…”

Section: Substrate Effects On Slb Formation Processmentioning

confidence: 99%

Substrate Effects on the Formation Process, Structure and Physicochemical Properties of Supported Lipid Bilayers

2012

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Supported lipid bilayers are artificial lipid bilayer membranes existing at the interface between solid substrates and aqueous solution. Surface structures and properties of the solid substrates affect the formation process, fluidity, two-dimensional structure and chemical activity of supported lipid bilayers, through the 1–2 nm thick water layer between the substrate and bilayer membrane. Even on SiO2/Si and mica surfaces, which are flat and biologically inert, and most widely used as the substrates for the supported lipid bilayers, cause differences in the structure and properties of the supported membranes. In this review, I summarize several examples of the effects of substrate structures and properties on an atomic and nanometer scales on the solid-supported lipid bilayers, including our recent reports.

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“…Solid-supported lipid membranes are frequently used to mimic phospholipid bio-interfaces, with widespread applications in biophysics and biomedical research. [1,2] Solid supported membranes can be prepared using a range of methods; [3][4][5][6] what is common in these that, depending on the intrinsic properties of the substrates, lipid molecules can assemble either into a bilayer or a monolayer on the support (with the exception when they remain in a pre-existing morphology as, e.g., vesicles or cubosomes). [4,7] The formation of phospholipid bilayers was achieved by various deposition methods on polar metal oxide surfaces such as titanium oxide, [8] aluminum oxide, [9] silicon oxide, [10] indium tin oxide, [11] as well as oxygen-plasma treated graphene [12] and even bare gold.…”

Section: Introductionmentioning

confidence: 99%

Formation of Alkanethiol Supported Hybrid Membranes Revisited

Zhi

Hasan²,

Mechler³

2018

Biotechnology Journal

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A phospholipid monolayer supported on an alkanethiol self-assembled monolayer (SAM) constitutes a supported hybrid membrane, a model of biological membranes optimized for electronic access through the underlying metal support surface. It is believed that phospholipids, when deposited from aqueous liposome suspension, spontaneously cover the alkanethiol-modified surface, owing to the reduction of surface free energy of the hydrophobic alkane surface exposed to the solution. However, the formation of the hybrid layer has to overcome significant energy barriers in rupturing the vesicle and "unzipping" the membrane leaflets; hence drivers of the spontaneous hybrid membrane formation are unclear. In this work, the authors studied the efficiency of the liposome deposition method to form hybrid membranes on octanethiol and hexadecanethiol SAMs in aqueous environment. Using quartz crystal microbalance to monitor the deposition process it was found that the hybrid membrane did not form spontaneously; the deposit was dominated by hemi-fused liposomes that can only be removed by applying osmotic stress. However, osmotic stress yielded a reproducible layer characterized by ≈-5Hz frequency change that is also confirmed by fluorescence microscopy imaging, irrespective of lipid concentration and the chain length of the SAMs. The frequency change is ≈20% of the frequency change expected for a tightly bound bilayer membrane, or 40% of a single leaflet, suggesting that the lipid layer is in a different conformation compared to a bilayer membrane: the acyl chains are most likely parallel to the SAM surface, likely due to strong hydrophobic interaction. Comparing these results to the literature it appears that the initial formation of hybrid membranes is inhibited by the ionic environment, while osmotic stress leads to the observed unique layer conformation.

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A universal method for planar lipid bilayer formation by freeze and thaw

Cited by 22 publications

References 52 publications

Versatile formation of supported lipid bilayers from bicellar mixtures of phospholipids and capric acid

Versatile formation of supported lipid bilayers from bicellar mixtures of phospholipids and capric acid

Substrate Effects on the Formation Process, Structure and Physicochemical Properties of Supported Lipid Bilayers

Formation of Alkanethiol Supported Hybrid Membranes Revisited

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