Potassium ion transport by valinomycin across a Hg-supported lipid bilayer

A biomimetic membrane consisting of a lipid bilayer tethered to a mercury electrode via a hydrophilic spacer was investigated in aqueous KCl by potential-step chronocoulometry and electrochemical impedance spectroscopy, both in the absence and in the presence of the ionophore valinomycin. Impedance spectra, recorded from 1 x 10(-2) to 1 x 10(5) Hz over a potential range of 0.8 V, are satisfactorily fitted to a series of four RC meshes, which are straightforwardly related to the different substructural elements of the biomimetic membrane. The frequency-independent resistances and conductances of both the lipid bilayer and the hydrophilic spacer show a maximum when plotted against the applied potential. This behavior is interpreted on the basis of a general approximate approach that applies the concepts of impedance spectroscopy to a model of the electrified interphase and to the kinetics of potassium ion transport assisted by valinomycin across the lipid bilayer.     

Gramicidin conducting dimers in lipid bilayers are stabilized by single-file ionic flux along them

Gramicidin D was incorporated in a biomimetic membrane consisting of a lipid bilayer tethered to a mercury electrode via a hydrophilic spacer, and its behavior was investigated in aqueous 0.1 M KCl by potential-step chronocoulometry and electrochemical impedance spectroscopy. The impedance spectra, recorded from 0.1 to 1 x 10(5) Hz over a potential range of 0.7 V, were fitted to a series of RC meshes, which were related to the different substructural elements of the biomimetic membrane. These impedance spectra were compared with those obtained by incorporating valinomycin, under otherwise identical conditions. The potential dependence of the stationary currents reported on bilayer lipid membranes by Bamberg and L?uger (Bamberg, E.; L?uger, P. J. Membrane Biol. 1973, 11, 177-194) as well as those extracted from potential-step chronocoulometric measurements was interpreted by relating the increase in gramicidin dimerization to a progressive increase in single-file K+ flux along the dimeric channels. An analogous approach was adopted in explaining the difference between the impedance spectra obtained with gramicidin D and those obtained with valinomycin. It is concluded that gramicidin has a low tendency to form dimers in the absence of ionic flux.     

Ion‐Channel Gating in Transmembrane Receptor Proteins: Functional Activity in Tethered Lipid Membranes

Through thiolipids a planar lipid bilayer (1) was immobilized on a gold support (2) for use as an electrode. This allows the detection of the ligand‐gating function of the natural transmembrane channel protein OmpF (3) reconstituted in the artificial membrane: the binding of a domain (4) of the toxin colicin N, observed by surface plasmon resonance, induces the blocking of the OmpF channel protein, as shown by impedance spectroscopy.     

Tethered bilayer lipid membranes self-assembled on mercury electrodes

In order to incorporate integral proteins in a functionally active state, metal-supported lipid bilayers must have a hydrophilic region interposed between the bilayer and the metal. This region is realized with a hydrophilic molecule terminating at one end with a sulfhydryl or disulfide group that anchors this "hydrophilic spacer" to the surface of a metal, such as gold or mercury. The other end of the hydrophilic spacer may be covalently linked to the polar head of a phospholipid molecule, giving rise to a supramolecule called "thiolipid" (TL). With respect to gold, mercury has the advantage of providing a defect-free and fluid surface to the self-assembling spacer. Hydrophilic spacers consisting of a polyethyleneoxy or a hexapeptide chain, as well as thiolipids derived from these spacers, were employed to fabricate mercury-supported lipid bilayers. The formation of a lipid bilayer on top of a self-assembled monolayer of a hydrophilic spacer, or of a single-lipid monolayer on top of a self-assembled monolayer of a thiolipid, was realized by simply immersing the coated mercury electrode into an aqueous solution across a lipid film previously spread on its surface at its spreading pressure. Particularly stable mercury-supported lipid bilayers were obtained by using thiolipids. The biomimetic properties of these lipid bilayers were tested by incorporating channel-forming polypeptides (gramicidin and melittin) and proteins (OmpF porin). The effect of the transmembrane potential on the function of these channels was estimated by using a simple electrostatic model of the mercury-solution interphase.     

Rapid screening of membrane protein activity: electrophysiological analysis of OmpF reconstituted in proteoliposomes

Solvent-free planar lipid bilayers were formed in an automatic manner by bursting of giant unilamellar vesicles (GUVs) after gentle suction application through micron-sized apertures in a borosilicate glass substrate. Incubation of GUVs with the purified ion channel protein of interest yielded proteoliposomes. These proteoliposomes allow for immediate recording of channel activity after GUV sealing. This approach reduces the time-consuming, laborious and sometimes difficult protein reconstitution processes normally performed after bilayer formation. Bilayer recordings are attractive for investigations of membrane proteins not accessible to patch clamp analysis, like e.g. proteins from organelles. In the presented work, we show the example of the outer membrane protein OmpF from Escherichia coli. We reconstituted OmpF in proteoliposomes and observed the characteristic trimeric conductance levels and the typical gating induced by pH and transmembrane voltage. Moreover, OmpF is the main entrance for beta-lactam antibiotics and we investigated translocation processes of antibiotics and modulation of OmpF by spermine. We suggest that the rapid formation of porin containing lipid bilayers is of potential for the efficient electrophysiological characterization of the OmpF protein, for studying membrane permeation processes and for the rapid screening of antibiotics.     

Electrochemical investigation of melittin reconstituted into a mercury-supported lipid bilayer

The channel-forming peptide melittin was incorporated into a biomimetic membrane consisting of a mercury electrode coated with a thiolipid monolayer, with a lipid monolayer self-assembled on top of it. The thiolipid consisted of a hydrophilic tetraethyleneoxy chain terminated at one end with a disulfide group, for anchoring to the mercury surface, and covalently linked at the other end to two diphytanyl chains, which formed a lipid bilayer with the overhanging lipid monolayer. The conductance of the lipid bilayer in contact with aqueous 0.1 M KCl was measured by electrochemical impedance spectroscopy over a frequency range from 1 x 10(-2) to 1 x 10(5) Hz and a potential range of 0.7 V for different compositions of the outer lipid monolayer. The conductance increases abruptly above the background level at sufficiently negative applied potentials, attaining a maximum value that increases with the composition of the outer monolayer in the order PC/chol (60:40) < PC < PC/SM/chol (59:15:26) < PS, with PC = phosphatidylcholine, chol = cholesterol, SM = sphingomyelin, and PS = phosphatidylserine. The higher the maximum conductance, the less negative the applied potential at which it is attained. This behavior is also discussed using a model of the electrified interphase.     

Formation and finite element analysis of tethered bilayer lipid structures

Rapid solvent exchange of an ethanolic solution of diphytanoyl phosphatidylcholine (DPhyPC) in the presence of a mixed self-assembled monolayer (SAM) [thiolipid/β-mercaptoethanol (βME) (3/7 mol/mol) on Au] shows a transition from densely packed tethered bilayer lipid membranes [(dp)tBLMs], to loosely packed tethered bilayer lipid membranes [(lp)tBLMs], and tethered bilayer liposome nanoparticles (tBLNs) with decreasing DPhyPC concentration. The tethered lipidic constructs in the aqueous medium were analyzed by atomic force microscopy (AFM) and electrochemical impedance spectroscopy (EIS). Finite element analysis (FEA) was applied to interpret spectral EIS features without referring to equivalent circuit modeling. Using structural data obtained earlier from neutron reflectometry and dielectric constants of lipid bilayers, we reproduced experimentally observed features of the electrochemical impedance (EI) spectra of complex surface constructs involving small pinhole defects, large membrane-free patches, and bound liposomes. We demonstrated by FEA that highly insulating (dp)tBLMs with low-defect density exhibit EI spectra in the shape of a perfect semicircle with or without low-frequency upward "tails" in the Cole-Cole representation. Such EI spectra were observed at DPhyPC concentrations of >5 × 10(-3) mol L(-1). While AFM was not able to visualize very small lateral defects in such films, EI spectra unambiguously signaled their presence by increased low frequency "tails". Using FEA we demonstrate that films with large diameter visible defects (>25 nm by AFM) produce EI spectral features consisting of two semicircles of comparable size. Such films were typically obtained at DPhyPC concentrations of <5 × 10(-3) mol L(-1). At DPhyPC concentrations of <1.0 × 10(-3) mol L(-1) the planar bilayer structures were replaced by ellipsoidal liposomes with diameters ranging from 50 to 500 nm as observed in AFM images. Despite the distinct surface morphology change, the EI curves exhibited two semicircle spectral features typical for the large size defects in planar tBLMs. FEA revealed that, to account for these EI features for bound liposome systems (50-500 nm diameter), one needs to assume much lower tBLM conductivities of the submembrane space, which separates the electrode surface and the phospholipid bilayer. Alternatively, FEA indicates that such features may also be observed on composite surfaces containing both bound liposomes and patches of planar bilayers. Triple semicircular features, observed in some of the experimental EI curves, were attributed to an increased complexity of the real tBLMs. The modeling demonstrated that such features are typical for heterogeneous tBLM surfaces containing large patches of different defectiveness levels. By integrating AFM, EIS, and FEA data, our work provides diagnostic criteria allowing the precise characterization of the properties and the morphology of surface supported bilayer systems.     

Controlling ionic conductivity in lipid polyelectrolyte composite capsules by cholesterol

The effect of cholesterol on the formation and properties of bilayer lipid membranes deposited on polyelectrolyte multilayered capsules was studied. The permeability of lipid/cholesterol coated capsules for NaCl was derived from osmotic response experiments and ranged from 1.45 x 10(-8) to 2.9 x 10(-8) m.s(-1), which corresponds to a lipid layer conductivity of (0.7-1.4) x 10(-8) S.m(-1). These conductivity values were in good agreement with the value of 0.8 x 10(-8) S.m(-1) obtained by electrorotation and were by 3 orders of magnitude lower than those found earlier for lipid layers on polyelectrolyte capsules in the absence of cholesterol.     

Electron transfer between ferrocene-modified Au/octadecanethiol/lipid BLM electrode and redox couples in solution

Bilayers incorporated with ferrocene consisting of self-assembled octadecanethiol and lipid monolayer on gold substrates were fabricated. Its electrochemical behaviors in solutions containing different redox couples were investigated by cyclic voltammetry and ac impedance. The transmembrane electron transfer reaction across octadecanethiol self-assembled film and an adsorbed phospholipid layer mediated by ferrocene have been observed in the solution of Fe(CN)6(3-/4-). The formal potential difference between mediator in bilayer lipid membrane (BLM) and redox couple in solution has a great impact on the transmembrane electron transfer behavior. The ferrocene-modified BLM electrodes might be useful for constructing a bilayer-based electrochemical current rectifying device.     

直流偏压对于在玻碳电极上双层类脂膜成膜过程的影响

应用循环伏安法和电化学阻抗谱研究了直流偏压对卵磷脂在玻碳电极表面自组装成膜过程及其结构的影响.实验发现:无论在正偏压还是负偏压条件下,卵磷脂在玻碳电极上均可组装成膜.施加正偏压时,由于玻碳电极表面所带的正电荷与卵磷脂端基之间的静电作用,使得卵磷脂在电极表面倾向于形成双层的类脂膜,并在0.4V偏压下电极阻抗达到最大值.继续增大电极正向偏压,s-BLM缺陷增加,以至趋于被击穿.提出了适宜的等效电路,并据此非线性拟合电极过程,求得部分阻抗的模型参数.研究发现:膜电容和电荷传递电阻呈现良好的互补效应.     

Formation of tethered bilayer lipid membranes on gold surfaces: QCM-Z and AFM study

Recently, tethered bilayer lipid membranes (tBLMs) have shown high potential as biomimetic systems due to their high stability and electrical properties, and have been used in applications ranging from membrane protein incorporation to biosensors. However, the kinetics of their formation remains largely uninvestigated. By using quartz crystal microbalance with impedance analysis (QCM-Z), we were able to monitor both the kinetics and viscoelastic properties of tether adsorption and vesicle fusion. Formation of the tether monolayer was shown to follow pseudo-first-order Langmuir kinetics with association and dissociation rate constants of 21.7 M-1 s(-1) and 7.43 x 10-6 s(-1), respectively. Moreover, the QCM-Z results indicate a rigid layer at the height of deposition, which then undergoes swelling as indicated by AFM. The deposition of vesicles to the tether layer also followed pseudo-first-order Langmuir kinetics with observed rate constants of 5.58 x 10(-2) and 2.41 x 10-2 s(-1) in water and buffer, respectively. Differential analysis of the QCM-Z data indicated deposition to be the fast kinetic step, with the rate-limiting steps being water release and fusion. Atomic force microscopy pictures taken complement the QCM-Z data, showing the major stages of tether adsorption and vesicle fusion, while providing a road map to successful tBLM formation.     

Electrostatic properties and macroscopic electrodiffusion in OmpF porin and mutants

The bacterial porin OmpF found in the outer membrane of E. coli is a wide channel, characterized by its poor selectivity and almost no ion specificity. It has an asymmetric structure, with relatively large entrances and a narrow constriction. By applying continuum electrostatic methods we determine the ionization states of titratable amino acid residues in the protein and calculate self-consistently the electric potential 3-D distribution within the channel. The average electrostatic properties are then represented by an effective fixed charge distribution along the pore which is the input for a macroscopic electrodiffusion model. The theoretical predictions agree with measurements performed under different salt gradients and pH. The sensitivity of reversal potential and conductance to the direction of the salt gradient and the solution pH is captured by the model. The theory is also able to explain the influence of the lipid membrane charge. The same methodology is satisfactorily applied to some OmpF mutants involving slight structural changes but a large number of net charges. The correlation found between atomic structure and ionic selectivity shows that the transport characteristics of wide channels like OmpF and its mutants are mainly regulated by the collective action of a large number of residues, rather than by the specific interactions of residues at particular locations.     

Loosely packed self-assembled monolayer of N-hexadecyl-3,6-di(p-mercaptophenylacetylene)carbazole on gold and its application in biomimetic membrane research

Dithiols of N-hexadecyl-3,6-di(p-mercaptophenylacetylene)carbazole (HDMC) have been synthesized and employed to form self-assembled monolayers (SAMs) on gold. One characteristic of the HDMC molecule is its peculiar molecular structure consisting of a large and rigid headgroup and a small and flexible alkyl-chain tail. HDMC adsorbates can attach to gold substrates by a strong Au-S bond with weak van der Waals interactions between the alkyl-chain tails, leading to a loosely packed hydrophobic SAM. In this way we can couple hybrid bilayer membranes (HBMs) to gold surfaces with more likeness to a cell bilayer than the conventional HBMs based on densely packed long-chain alkanethiol SAMs. The insulating properties and stability of the HDMC monolayer as well as the HDMC/lipid bilayer on gold have been investigated by electrochemical techniques including cyclic voltammetry and impedance spectroscopy. To test whether the quality of the bilayer is sufficiently high for biomimetic research, we incorporated the pore-forming protein alpha-hemolysin) and the horseradish peroxidase into the bilayers, respectively. Experimental results demonstrated that this type of loosely packed hydrophobic SAM has great potential in biomimetic bilayer research and biosensor application.     

Surface response methodology for the study of supported membrane formation

We report on the investigations of the formation of the tethered lipid bilayer by vesicle deposition on amine-functionalized surfaces. The tethered bilayer was created by the deposition of egg-PC vesicles containing 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-poly-(ethyleneglycol)-N-hydroxysuccinimide as anchoring molecules on an amine-coated surface. This approach is an easy route for the formation of a biomimetic-supported membrane. A Doelhert experimental design was applied to determine the conditions leading to the formation of a continuous and defect-free tethered bilayer on different surfaces (gold and glass). Doehlert designs allow modeling of the experimental responses by second-order polynomial equations as a function of experimental factors. Four factors expected to influence bilayer formation were studied: the lipid concentration in the vesicle suspension, the mass percentage of anchoring molecules in the vesicles, the contact time between the vesicles and the surface, and the resting time of the membrane after buffer rinse. The optimization of the membrane preparation parameters was achieved by monitoring lipid assembly formation using surface plasmon resonance spectroscopy on gold and by fluorescence recovery after photobleaching on glass. Three characteristic responses were systematically measured: the bilayer thickness, the lipid diffusion coefficient, and the lipid mobile fraction. The simultaneous inspection of the three characteristics revealed that a restricted experimental domain leads to properties that are in accordance with a bilayer presence. The factors of this domain are a lipid concentration from 0.1 to 1 mg/mL, 4-8% of anchoring molecules in the vesicles, 1-4 h of contact time between vesicles and surface, and 21-24 h of resting time after buffer rinse. Under these conditions, a membrane having a lipid mass per surface between 545 +/- 5 and 590 +/- 10 ng/cm2, a diffusion coefficient of between 2.5 +/- 0.3 x 10(-8) and 3.60 +/- 0.5 x 10(-8) cm2/s, and a mobile fraction between 94 +/- 2 and 99 +/- 1% was formed. These findings were confirmed by atomic force microscopy observations, which showed the presence of a continuous and homogeneous bilayer in the determined experimental domain. This formation procedure presents many advantages; it provides an easily obtainable biomimetic membrane model for proteins studies and offers a versatile tethered bilayer because it can be adapted easily to various types of supports.     

杂多酸K7Fe^3^+P2W17O62H2对硫醇-磷脂杂化双层膜通透性的影响

利用涂抹冷冻法制备了硫醇-磷脂杂化双层膜,采用循环伏安和交流阻抗方法,研究了硫醇-磷脂杂化双层膜与杂多酸K7Fe^3^+P2W17O62H2作用前后通透性的变化,发现该种杂多酸能够诱导硫醇-磷脂杂化双层膜产生一些孔洞,降低了膜电阻,增加了膜电容,也增加了探针Fe(CN)^3-/4-~6与电极的电子传递。同时对产生该现象的机理进行了初步的探讨。     

Molecular dynamics simulations of DiI-C18(3) in a DPPC lipid bilayer

We performed a 40 ns simulation of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI-C18(3)) in a 1,2-dipalmitoyl-sn-glycero-3-phosphatidyl choline (DPPC) bilayer in order to facilitate interpretation of lipid dynamics and membrane structure from fluorescence lifetime, anisotropy, and fluorescence correlations spectroscopy (FCS). Incorporation of DiI of 1.6 to 3.2 mol% induced negligible changes in area per lipid but detectable increases in bilayer thickness, each of which are indicators of membrane structural perturbation. The DiI chromophore angle was 77 +/- 17 degrees with respect to the bilayer normal, consistent with rotational diffusion inferred from polarization studies. The DiI headgroup was located 0.63 nm below the lipid head group-water interface, a novel result in contrast to some popular cartoon representations of DiI but consistent with DiI's increase in quantum yield when incorporated into lipid bilayers. Importantly, the fast component of rotational anisotropy matched published experimental results demonstrating that sufficient free volume exists at the sub-interfacial region to support fast rotations. Simulations with non-charged DiI head groups exhibited DiI flip-flop, demonstrating that the positively-charged chromophore stabilizes the orientation and location of DiI in a single monolayer. DiI induced detectable changes in interfacial properties of water ordering, electrostatic potential, and changes in P-N vector orientation of DPPC lipids. The diffusion coefficient of DiI (9.7 +/- 0.02 x 10(-8) cm2 s(-1)) was similar to the diffusion of DPPC molecules (10.7 +/- 0.04 x 10(-8) cm2 s(-1)), supporting the conclusion that DiI dynamics reflect lipid dynamics. These results provide the first atomistic level insight into DiI dynamics, results essential in elucidating lipid dynamics through single molecule fluorescence studies.     

Immobilizing single lipid and channel molecules in artificial lipid bilayers with annexin A5

The effects of annexin A5 on the lateral diffusion of single-molecule lipids and single-molecule proteins were studied in an artificial lipid bilayer membrane. Annexin A5 is a member of the annexin superfamily, which binds preferentially to anionic phospholipids in a Ca2+-dependent manner. In this report, we were able to directly monitor single BODIPY 1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine (DHPE) and ryanodine receptor type 2 (RyR2) labeled with Cy5 molecules in lipid bilayers containing phosphatidylserine (PS) by using fluorescence microscopy. The diffusion coefficients were calculated at various annexin A5 concentrations. The diffusion coefficients of BODIPY-DHPE and Cy5-RyR2 in the absence of annexin A5 were 4.81 x 10(-8) cm(2)/s and 2.13 x 10(-8) cm(2)/s, respectively. In the presence of 1 microM annexin A5, the diffusion coefficients of BODIPY-DHPE and Cy5-RyR2 were 2.2 x 10(-10) cm(2)/s and 9.5 x 10(-11) cm(2)/s, respectively. Overall, 1 microM of annexin A5 was sufficient to induce a 200-fold decrease in the lateral diffusion coefficient. Additionally, we performed electrophysiological examinations and determined that annexin A5 has little effect on the function of RyR2. This means that annexin A5 can be used to immobilize RyR2 in a lipid bilayer when imaging and analyzing RyR2.     

Colloid stability of lipid/polyelectrolyte decorated latex

The colloid stability of supramolecular assemblies composed of the synthetic cationic lipid dioctadecyldimethylammonium bromide (DODAB) on carboxymethyl cellulose (CMC) supported on polystyrene amidine (PSA) microspheres was evaluated via turbidimetry kinetics, dynamic light scattering for particle sizing, zeta-potential analysis, and determination of DODAB adsorption on CMC-covered particles. At 0.1 g L(-1) CMC and 2 x 10(11) PSA particles/mL, CMC did not induce significant particle flocculation, and a vast majority of CMC-covered single particles were present in the dispersion so that this was the condition chosen for determining DODAB concentration (C) effects on particle size and zeta potentials. At 0.35 mM DODAB, charge neutralization, maximal size, and visible precipitation indicated extensive flocculation and minimal colloid stability for the DODAB/CMC/PSA assembly. At 0.1 g L(-1) CMC, isotherms of high affinity for DODAB adsorption on CMC-covered particles presented a plateau at a limiting adsorption of 700 x 10(17) DODAB molecules adsorbed per square meter PSA which was well above bilayer deposition on a smooth particle surface. The polyelectrolyte layer on hydrophobic particles was swelled and fluffy (ca. 11-nm hydrodynamic thickness), and maximal adsorption of DODAB lipid onto this layer produced a compressed composite cationic film with 20 mV of zeta potential and about 10-nm mean thickness. The assembly of cationic lipid/CMC layer/polymeric particle was stable only well above charge neutralization of the polyelectrolyte by the cationic lipid, at relatively large lipid concentrations (at and above 1 mM DODAB) with charge neutralization leading to extensive particle aggregation.     

Mott-Schottky Analysis and Impedance Spectroscopy of TiO2/6T and ZnO/6T devices

Schottky junctions have been realized by evaporating gold spots on top of sexithiophen (6T), which is deposited on TiO 2 or ZnO with e-beam and spray pyrolysis. Using Mott-Schottky analysis of 6T/TiO2 and 6T/ZnO devices acceptor densities of 4.5x10(16) and 3.7x10(16) cm(-3) are obtained, respectively. For 6T/TiO2 deposited with the e-beam evaporation a conductivity of 9x10(-8) S cm(-1) and a charge carrier mobility of 1.2x10(-5) cm2/V s is found. Impedance spectroscopy is used to model the sample response in detail in terms of resistances and capacitances. An equivalent circuit is derived from the impedance measurements. The high-frequency data are analyzed in terms of the space-charge capacitance. In these frequencies shallow acceptor states dominate the heterojunction time constant. The high-frequency RC time constant is 8 micros. Deep acceptor states are represented by a resistance and a CPE connected in series. The equivalent circuit is validated in the potential range (from -1.2 to 0.8 V) for 6T/ZnO obtained with spray pyrolysis.     

Monte Carlo study of lipid membranes: Simulation of diparmitoylphosphatidylcholine bilayers in gel and liquid-crystalline phases

The statistical properties of the bilayer membranes of diparmitoylphosphatidylcholine (DPPC) in the gel and liquid-crystal phases were studied by Monte Carlo (MC) simulation using potential functions of the Lennard-Jones, the simple Coulomb, and the bond torsion. The simulation was undertaken on a two-dimensional periodic condition imposed on the bilayer model consisting of faithfully described molecules. The structure and ordering of the model bilayers accorded well with experiments, and the segment order parameters were in agreement with those of the nuclear magnetic resonance (NMR) experiments. The two kinds of lipid chains of DPPC do not equivalently behave in the bilayers, and chain 2 has lower ordering than chain 1. The order parameters of the first eight segments of chain 2 in the liquid-crystal model are particularly small and are roughly constant. From electron density analysis, it has been observed that the liquid-crystal bilayer has about one excess water molecule per one lipid molecule in comparison with the gel bilayer. The energy difference between the two bilayer models, taking account of the water contribution, is consistent with the latent heat of the phase transition. © 1995 by John Wiley & Sons, Inc.