Solid-Supported Bilayer Lipid Membranes

With only a few exceptions, metal-supported biomimetic membranes consist of a more or less complex architecture that includes a lipid bilayer. In order of increasing complexity, they can be classified into solid-supported bilayer lipid membranes (sBLMs), tethered bilayer lipid membranes (tBLMs), polymer-cushioned bilayer lipid membranes (pBLMs), S-layer stabilized bilayer lipid membranes (ssBLMs), and protein-tethered bilayer hpid membranes (ptBLMs). 

In a subsequent study [30], cholesteryl-substituted 18C6 derivative 11 and diaza [18]crown-6 12 (Scheme 6) were used to create solid-supported bilayer lipids. The liquid crystalline crown derivatives 11,12 were dissolved in chloroform and mixed with squalene or squalene saturated with cholesterol. The solid-supported bilayers were prepared in freshly cut stainless steel wires. A 10-4 to 10-1 mol L-1 solution of MCI (M = Li, Na, K, Rb, Cs) or MgCl2 was used as aqueous phase. Measurement of the membrane potential revealed a Nemst response to the concentration of M+ in solution. It was possible to differentiate between the different cations which might be used for the preparation of new ion sensors. For the detection of K+ and Rb+, aza crown derivative 12 proved to be the most selective. A problem was the presence of traces of Fe2+/3+ that made the measurements difficult. It was also not... 

Seitz M ef al 1998 Formation of tethered supported bilayers via membrane-inserting reactive lipids Thin Solid Films 327-9 767-71... 

Different other attractive way to modify the active electrode of amperometric sensors, besides deposition of polymers or electropolymerization, is formation of self-assembled structures, e.g., self-assembled monolayers (SAM) on solid supports 88 89 or bilayer lipid membranes (BLM) on various types of support.90 Both types of theses structures can either induce selectivity of sensor to particular analytes and... 

Fig. 12 Neutron reflectometry (NR) data of the polyelectrolyte multilayer (4 PSS/4 PAH) - coated solid support without lipid bilayer (A), and with a DMPC/DMPG (10 1) mixed membrane on top (C). The curves are shifted relative to each other for clarity. Solid lines represent model calculations of the data with scattering length densities, b/V, corresponding to the blank multilayer support (A), and to the tethered bilayer plus a nonspecific top layer (C), as given in the inset. The dotted line (B) represents a simulation of a lipid bilayer without an additional nonspecific layer on top...

BLMs, especially self-assembled bilayer lipid membranes on solid supports (s-BLMs), first reported in 1989, have been used in the last several years as lipid bilayer-based biosensors. 

Various planar membrane models have been developed, either for fundamental studies or for translational applications monolayers at the air-water interface, freestanding films in solution, solid supported membranes, and membranes on a porous solid support. Planar biomimetic membranes based on amphiphilic block copolymers are important artificial systems often used to mimic natural membranes. Their advantages, compared to artificial lipid membranes, are their improved stability and the possibility of chemically tailoring their structures. The simplest model of such a planar membrane is a monolayer at the air-water interface, formed when amphiphilic molecules are spread on water. As cell membrane models, it is more common to use free-standing membranes in which both sides of the membrane are accessible to water or buffer, and thus a bilayer is formed. The disadvantage of these two membrane models is the lack of stability, which can be overcome by the development of a solid supported membrane model. Characterization of such planar membranes can be challenging and several techniques, such as AFM, quartz crystal microbalance (QCM), infrared (IR) spectroscopy, confocal laser scan microscopy (CLSM), electrophoretic mobility, surface plasmon resonance (SPR), contact angle, ellipsometry, electrochemical impedance spectroscopy (EIS), patch clamp, or X-ray electron spectroscopy (XPS) have been used to characterize their... 

Here, we discuss a solid-state 19F-NMR approach that has been developed for structural studies of MAPs in lipid bilayers, and how this can be translated to measurements in native biomembranes. We review the essentials of the methodology and discuss key objectives in the practice of 19F-labelling of peptides. Furthermore, the preparation of macroscopically oriented biomembranes on solid supports is discussed in the context of other membrane models. Two native biomembrane systems are presented as examples human erythrocyte ghosts as representatives of eukaryotic cell membranes, and protoplasts from Micrococcus luteus as membranes... 

For the orientation-based structure analysis of MAPs, uniformly oriented lipid bilayers are typically prepared on solid supports as illustrated in Fig. 2 [23, 47, 55]. These mechanically oriented membranes are advantageous for static ssNMR experiments, as they provide a robust way to orient a sample with any desired lipid composition, peptide concentration, and at any desired temperature. The lipids... 

Fig. 5 Membrane models for NMR structure analysis, (a) An isotropic detergent micelle (left) is compared to the dimensions of lipid bilayers (right), (b) Macroscopically oriented membrane samples can be prepared on solid support, as nanodiscs, or as magnetically oriented bicelles. (c) Nomenclature and variability of liposomes small (SUV, 20-40 nm), intermediate (IUV, 40-60 nm), large (LUV, 100-400 nm), and giant unilamellar vesicles (GUV, 1 pm) multi-lamellar (MLV), oligo-lamellar (OLV) and highly heterogeneous multi-oligo-lamellar vesicles (MOLV)...

The use of solid-supported lipid membranes (SSLM) to measure membrane affinity was recently reported by Loidl-Stahlhofen (2001a, 2001b). To produce solid-supported lipid membranes a single phospholipid bilayer membrane is non covalently... 

Figure 3 Methods for supported bilayer formation and membrane protein reconstitution, (a) and (b) LB/LS method. A lipid monolayer is spread at the air-water interface of a Langmuir trough and transferred to a solid substrate while keeping the surface pressure constant. A second monolayer is transferred by horizontal apposition of the first transferred monolayer and collection of a counter-piece with spacers, (c) Direct VF method. Membrane vesicles are flown into a chamber with a clean surface substrate on top. After about an hour of incubation, they form a supported bilayer on the substrate and excess vesicles are flushed out. (d) LB/VF method. The procedures depicted in panels (a) and (c) are combined leading to an asymmetric bilayer with an asymmetric protein distribution. Although this method can also be performed without a polymer, it is shown here with the polymer transferred during the LB step.

Methodologies for the fabrication of biomimetic membranes vary somewhat from one biomimetic membrane to another. However, a number of experimental procedures for the formation of lipid monolayers and bilayers on solid supports are common to several biomimetic membranes. The most popular procedures are vesicle fusion, Langmuir-Blodgett and Langmuir-Schaefer transfers, and rapid solvent exchange. The formation of lipid monolayers and bilayers on gold and... 

Figure 4.11 Schematic of S-layer stabilized solid supported lipid membranes, (a) S-layer directly recrystallized on gold, with a lipid bilayer on top. (b) Same as (a), with an additional S-layer recrystallized on top of the lipid bilayer, (c) Thiolated SCWPs directly bound to gold and interacting with an S-layer, with a lipid bilayer on top. (d) Same as (c).

Many questions pertaining to membrane processes in general and ligand/membrane receptor interactions in particular can be addressed by a novel model membrane system, i.e., polymer-supported or polymer-tethered lipid bilayers [12,14], The basic structural unit for this sensor platform is the tethered lipid bilayer membrane [16] displayed in Fig. 2D. The essential architectural elements of this supramolecular assembly include the solid support, e.g., an optical or electrical transducer (device), the polymeric tether system which provides the partial covalent and, hence, very stable attachment of the whole membrane to the substrate surface, and the fluid lipid bilayer, functionalized if needed by embedded proteins. 

Fig. 2 The construction of a polymer-cushioned lipid bilayer membrane. (A) Architecture constructed in a sequential way first, onto the functionalized substrate a polymer layer (cushion) is deposited by adsorption from solution and covalent binding, followed by the (partial) covalent attachment of a lipid monolayer containing some anchor lipids as reactive elements (B) able to couple the whole monolayer to the polymer cushion. (C) Alternatively, a lipopolymer monolayer, organized, e.g., at the water-air interface can be co-spread with regular low-mass amphiphiles and then transferred as a mixed monolayer onto a solid support, prefunctionalized with reactive groups, able to bind covalently to the polymer chains of the lipopolymer molecules, (B). (D) By a fusion step (or a Langmuir Schaefer transfer) the distal lipid monolayer completes the polymer-tethered membrane architecture...

Among the various membrane properties that might need to be optimized for a particular application of the tethered membrane architecture and/or of any incorporated proteins we discuss only two key performance parameters, i.e., (1) the ability of the tethering system to swell by the up-take of a sufficient amount of water into the interstitial space between the lipid bilayer and the solid support, and (2) the high lateral mobility of the individual lipid molecules in the two opposing leaflets of the bilayer membrane. 

---