Electrochemical and PM-IRRAS studies of the effect of the static electric field on the structure of the DMPC bilayer supported at a Au(111) electrode surface

Differential capacity, charge density measurements, and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) were employed to study the fusion of small unilamellar vesicles of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) on a Au(111) electrode surface. The differential capacity and charge density data showed that the vesicles fuse onto the gold surface at charge densities between -10 microC/cm(2) < sigma(M) < 10 microC/cm(2) to form a bilayer. When sigma(M) < -10 microC/cm(2), the film is detached from the surface but it remains in close proximity to the surface. PM-IRRAS experiments provided IR spectra for the bilayer in the adsorbed and the desorbed state. Ab initio normal coordinate calculations were performed to assist interpretation of the IR spectra. The IR bands were analyzed quantitatively, and this analysis provided information concerning the conformation and orientation of the acyl chains and the polar head region of the DMPC molecule. The orientation of the chains, hydration, and conformation of the headgroup of the DMPC molecule strongly depend on the electrode potential.     

Electrochemical and FTIR studies of -phenylalanine adsorption at the Au(111) electrode

The adsorption of -phenylalanine (Phe) at the Au(111) electrode surface has been studied using electrochemical techniques and subtractively normalized interfacial Fourier transform infrared (SNIFTIR) techniques. The electrochemical measurements of cyclic voltammetry, differential capacity and chronocoulometry were used to determine Gibbs energies of adsorption and the reference (E₁) and sample (E₂) potentials to be used in the spectroscopic measurements. The vibrational spectra have been used to determine: (i) the orientation of the molecule at the surface as a function of potential; (ii) the dependence of the band intensity on the surface coverage; (iii) the character of surface coordination, and (iv) the oxidation of adsorbed Phe molecules at positive potentials. The adsorption of Phe is characterized by ΔG values ranging from −18 to −37 kJ mol⁻¹ that are characteristic for a weak chemisorption of small aromatic molecules. The electrochemical and SNIFTIR measurements indicated that adsorbed Phe molecules change orientation as a function of applied potential. At the negatively charged surface Phe is predominantly adsorbed in the neutral form of the amino acid. At potentials positive to the pzc, adsorption occurs predominantly in the zwitterionic form with the ---COO⁻ group directed towards the surface and the ammonium group towards the solution. At more positive potentials electrocatalytic oxidation of Phe occurs and is marked by the appearance of the CO₂ asymmetric stretch band in the FTIR spectrum. Thus, relative to pzc, Phe is weakly chemisorbed at negative potentials, changes orientation at potentials close to the pzc and is oxidized at positive potentials.     

Electrochemical and PM-IRRAS studies of potential controlled transformations of phospholipid layers on Au(111) electrodes

Chronocoulometry and photon polarisation modulation infrared reflection absorption spectroscopy (PM-IRRAS) have been employed to study the fusion of dimyristoylphosphatidylcholine (DMPC) vesicles onto a Au(111) electrode surface. The results show that fusion of the vesicles is controlled by the electrode potential or charge at the electrode surface (sigmaM). At charge densities of -15 microC cm(-2) < sigmaM < 0 microC cm(-2), DMPC vesicles fuse to form a condensed film. When sigmaM < -15 microC cm(-2), de-wetting of the film from the electrode surface occurs. The film is detached from the electrode surface; however, phospholipid molecules remain in its close proximity in an ad-vesicle state. The state of the film can be conveniently changed by adjustment of the potential applied to the gold electrode. PM-IRRAS experiments demonstrated that the potential-controlled transitions between various DMPC states proceed without conformational changes and changes in the packing of the acyl chains of DMPC molecules. However, a remarkable change in the tilt angle of the acyl chains with respect to the surface normal occurs when ad-vesicles spread to form a film at the gold surface. When the bilayer is formed at the gold surface, the acyl chains of DMPC molecules are significantly tilted. The IR spectra have also demonstrated a pronounced change in the hydration of the polar head region that accompanies the spreading of ad-vesicles into the film. For the film deposited at the electrode surface, the infrared results showed that the temperature-controlled phase transition from the gel state to the liquid crystalline state occurs within the same temperature range as that observed for aqueous solutions of vesicles. The results presented in this work show that PM-FTIR spectroscopy, in combination with electrochemical techniques, is an extremely powerful tool for the study of the structure of model membrane systems at electrode surfaces.     

STM studies of fusion of cholesterol suspensions and mixed 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC)/cholesterol vesicles onto a Au(111) electrode surface

Electrochemical scanning tunneling microscopy (EC-STM) has been applied to study the structure of the film formed by fusion of cholesterol suspensions and mixed dimyristoylphosphatidylcholine (DMPC)/cholesterol vesicles on a Au(111) electrode surface. It has been demonstrated that cholesterol molecules assemble at the gold support into several structures templated by the crystallography of the metal surface and involving flat or edge-on adsorbed molecules. Studies of the film formed by fusion of mixed DMPC/cholesterol vesicles revealed that ordered domains of either pure DMPC or pure cholesterol were formed. These results indicate that, at the metal surface, the molecules released by the rupture of a vesicle initially self-assemble into a well-ordered monolayer. The self-assembly is controlled by the hydrocarbon skeleton-metal surface interaction. In the case of mixed DMPC/cholesterol vesicles, the molecule-metal interactions induce segregation of the two components into single component domains. However, the molecule-metal interaction induced monolayer is a transient phenomenon. When more molecules accumulate at the surface, the molecule-molecule interactions dominate the assembly, and the monolayer is transformed into a bilayer.     

Layer-by-layer PMIRRAS characterization of DMPC bilayers deposited on a Au111 electrode surface

A combination of Langmuir-Blodgett and Langmuir-Schaefer techniques was employed to deposit 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers at a gold electrode surface. One leaflet consisted of hydrogen-substituted acyl chains, and the second leaflet was composed of molecules with deuterium-substituted acyl chains. This architecture allowed for layer-by-layer analysis of the structure of the bilayer. Photon polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS) was used to determine the conformation and orientation of the acyl chains of DMPC molecules in the individual leaflets as a function of the potential applied to the gold electrode. The bilayer is adsorbed onto the metal surface when the field applied to the membrane does not exceed approximately 108 V/m. When adsorbed, the bottom leaflet is in contact with a hydrophobic metal surface, and the top leaflet is interacting with the aqueous solution. The asymmetry of the environment has an effect on the orientation of the DMPC molecules in each leaflet. The tilt angle of the acyl chains of the DMPC molecules in the bottom leaflet that is in contact with the gold is approximately 10 degrees smaller than that observed for the top leaflet that is exposed to the solution. These studies provide direct evidence that the structure of a phospholipid bilayer deposited at an electrode surface is affected by interaction with the metal.     

Electric field driven changes of a gramicidin containing lipid bilayer supported on a Au(111) surface

Langmuir-Blodgett and Langmuir-Schaeffer methods were employed to deposit a mixed bilayer consisting of 90% of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and 10% of gramicidin (GD), a short 15 residue ion channel forming peptide, onto a Au(111) electrode surface. This architecture allowed us to investigate the effect of the electrostatic potential applied to the electrode on the orientation and conformation of DMPC molecules in the bilayer containing the ion channel. The charge density data were determined from chronocoulometry experiments. The electric field and the potential across the membrane were determined through the use of charge density curves. The magnitudes of potentials across the gold-supported biomimetic membrane were comparable to the transmembrane potential acting on a natural membrane. The information regarding the orientation and conformation of DMPC and GD molecules in the bilayer was obtained from photon polarization modulation infrared reflection absorption spectroscopy (PMIRRAS) measurements. The results show that the bilayer is adsorbed, in direct contact with the metal surface, when the potential across the interface is more positive than -0.4 V and is lifted from the gold surface when the potential across the interface is more negative than -0.4 V. This change in the state of the bilayer has a significant impact on the orientation and conformation of the phospholipid and gramicidin molecules. The potential induced changes in the membrane containing peptide were compared to the changes in the structure of the pure DMPC bilayer determined in earlier studies.     

Self-assembly of phospholipid molecules at a Au(111) electrode surface

We described the first scanning tunneling microscopy study of spreading unilamellar vesicles of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) at a Au(111) electrode surface. At the initial stage of the film formation, the molecular resolution images revealed that DMPC molecules are adsorbed flat with the acyl chains oriented parallel to the surface. The molecules assemble into double rows by aligning the acyl chains in the nearest neighbor direction of the reconstructed Au(111) surface and assuming a 90 +/- 10 degrees angle with respect to line of the molecular row. After approximately 30 min, this film is transformed into a hemimicellar state with long rows characteristic for the formation of hemicylindrical surface micelles. At hydrophilic surfaces such as glass, spreading of vesicles involves adsorption, rupture, and sliding of a single bilayer on a lubricating film of the solvent. We have provided the first evidence that a different mechanism is involved in spreading the vesicles at gold. The molecules released by rupture of vesicles self-assemble into an ordered film, and the assembly is controlled by the chain-substrate interaction.     

金(111)电极表面浮动磷脂双层膜的原位偏振红外反射光谱研究(英文)

应用电化学原位偏振红外反射光谱法研究了构建于金(111)电极表面的浮动磷脂双层膜.金电极表面先自组装一层巯基葡萄糖单层来增加表面的亲水性,浮动磷脂双层膜通过LB-LS技术构建在巯基葡萄糖单层上.双层膜由双肉豆蔻磷脂酰胆碱(DMPC)、胆固醇和神经节苷脂GM1构成.GM1分子中的糖链可以物理吸附在巯基葡萄糖表面,在双层膜和基底间形成一个富含水的隔层.红外光谱表明浮动双层膜中的DMPC分子比传统的支撑双层膜中的DMPC分子有更强的水合作用,证实了双层膜和基底间水层的存在.该浮动双层膜更接近于实际的生物膜体系,并且在金电极表面有宽的电位区间,非常适于进一步的离子通道蛋白质研究.     

Infrared studies of the potential controlled adsorption of sodium dodecyl sulfate at the Au(111) electrode surface

Quantitative subtractively normalized interfacial Fourier transform infrared reflection spectroscopy (SNIFTIRS) was used to determine the conformation and orientation of sodium dodecyl sulfate (SDS) molecules adsorbed at the single crystal Au(111) surface. The SDS molecules form a hemimicellar/hemicylindrical (phase I) structure for the range of potentials between -200 ≤ E < 450 mV and condensed (phase II) film for electrode potentials ≥500 mV vs Ag/AgCl. The SNIFTIRS measurements indicate that the alkyl chains within the two adsorbed states of SDS film are in the liquid-crystalline state rather than the gel state. However, the sulfate headgroup is in an oriented state in phase I and is disordered in phase II. The newly acquired SNIFTIR spectroscopy measurements were coupled with previous electrochemical, atomic force microscopy, and neutron reflectivity data to improve the current existing models of the SDS film adsorbed on the Au(111) surface. The IR data support the existence of a hemicylindrical film for SDS molecules adsorbed at the Au(111) surface in phase I and suggest that the structure of the condensed film in phase II can be more accurately modeled by a disordered bilayer.     

Electrochemical Studies of the Benzoate Adsorption on Au (111) Electrode

Cyclic voltammetry (CV), differential capacity (DC), and charge densitymeasurements have been employed to study the benzoate (BZ) adsorption at the Au(111)electrode surface. Thermodynamic analysis of charge density (_M) data has beenperformed to describe the properties of the adsorbed benzoate ion. The Gibbsexcess , Gibbs energy of adsorption G, and the number of electrons flowingto the interface per adsorbed benzoate ion at a constant potential (electrosorptionvalency) and at a constant bulk concentration of the benzoate (reciprocal of theEsin—Markov coefficient) have been determined. The results demonstrate thatalthough benzoate adsorption starts at negative charge densities, it takes placepredominantly at a positively charged surface. At the most positive potentials,the surface concentration of benzoate attains a limiting value of about 7.3×10^–10mol-cm^–2, which is independent of the bulk benzoate concentration. This valueis consistent with packing density corresponding to a closed-packed monolayerof vertically adsorbed benzoate molecules. At negative charge densities, benzoateassumes a flat (-bonded) surface coordination. The surface coordination ofbenzoate changes, by moving from a negatively to positively charged surface.At the negatively charged surface, the electrosorption bond is quite polar. Thepolarity of the chemisorption bond is significantly reduced due either to a chargetransfer or a screening of the charge on the anion by the charge on the metal.     

Electrochemical redox control of ferrocene using a supramolecular assembly of ferrocene-linked C(60) derivative and metallooctaethylporphyrin array on a Au(111) electrode

Supramolecular assembled layers of ferrocene-linked C(60) derivative (C(60)Fc) and various metal ions coordinated to octaethylporphyrin (MOEP) were formed on the surface of a Au(111) single-crystal electrode by immersing the Au substrate successively into a benzene solution containing MOEP and one containing C(60)Fc molecules. The MOEPs used were zinc(II) (ZnOEP), cobalt(II) (CoOEP), copper(II) (CuOEP), and iron(III) chloride (FeClOEP) of OEP (2,3,7,8,12,13,17,18-octaethyl-21H,23H-porphine). The molecules of C(60)Fc directly attached to the Au(111) electrode showed poorly defined electrochemical redox response, whereas a clear electrochemical redox reaction of the ferrocene group in the C(60)Fc molecule was observed at 0.78 V versus reversible hydrogen electrode on ZnOEP, CoOEP, and CuOEP adlayers, but not on the FeClOEP adlayer. Adlattices of the underlying layer and the top layer of C(60)Fc were determined by in situ scanning tunneling microscopy. Adlayer structures of MOEP were independent of the central metal ion; that is, MOEP molecules were arranged hexagonally with two different orientations. Highly ordered C(60)Fc arrays were formed with 1:1 composition on the ZnOEP-, CoOEP-, and CuOEP-modified Au(111) surface, whereas a disordered structure of C(60)Fc was found on the FeClOEP-modified Au(111) surface. The presence of Cl ligand was found to prevent the formation of supramolecularly assembled layers with C(60)Fc molecules, resulting in an ill-defined unclear electrochemical response of the Fc group. The well-defined electrochemical response of the Fc group in C(60)Fc was clearly due to the control of orientation of C(60)Fc molecules.     

Spectroelectrochemical studies of bilayers of phospholipids in gel and liquid state on Au(111) electrode surface

Differential capacity, chronocoulometry and Polarization Modulation Fourier Transform Infrared Reflection Absorption Spectroscopy (PM FTIRRAS) were employed to investigate spreading of small unilamellar vesicles (SUVs) of DOPC and DMPC onto a Au(111) electrode surface. The electrochemical experiments demonstrated that vesicles fuse onto the electrode surface and at E>-0.5V (SSCE) or at charge densities -10-0.5 V (SSCE), the tilt angle increases to approximately 42 degrees. The increase of the tilt angle is discussed in terms of a change in the packing of the polar head of the phospholipids molecules in the bilayer adsorbed at the electrode surface.     

Au单晶表面氟表面活性剂FSN自组装膜的电化学行为

研究Au(111)和Au(100)表面非离子型氟表面活性剂FSN自组装膜的电化学行为.电化学扫描隧道显微术和循环伏安法测试表明,在0～0.8 V电位区间,FSN自组装膜未发生氧化还原,均一性好,可稳定地存在于电极表面,并显著抑制硫酸根离子在电极表面的吸附和Au单晶表面的重构.在FSN自组装膜Au单晶电极的初始氧化阶段,Au(111)表面有少量突起,而Au(100)表面呈现台阶剧烈变化,但FSN自组装膜的吸附结构没有改变.与Au(100)表面相比,Au(111)表面形成的FSN自组装膜可更有效地抑制Au表面的氧化.     

Electrochemical detection of cytosine and 5-methylcytosine on Au(111) surfaces

In this communication we report a voltammetric study of the adsorption–desorption of cytosine (C) and methylcytosine (mC) on well-defined gold (Au) electrodes. The voltammetric measurements clearly indicate that these processes are extremely sensitive to the Au surface structure and in particular to the presence of (111) surface domains. Interestingly, on Au(111) surfaces, a linear correlation between the C and mC concentrations (logarithm scale) and the peak potential of the main voltammetric feature is found. In addition, in the simultaneous presence of both molecules, mC governs the electrochemical response, which has allowed its accurate quantification in C-mC mixtures. In situ FTIR spectroscopic measurements have been carried out to deepen on this mC electrochemical sensitivity. This research may contribute to the future development of an electrochemical sensor for the determination of the degree of methylation in DNA.     

Electrochemical studies of technetium at a mercury electrode

The electrochemistry of technetium was studied by polarography, cyclic voltammetry and coulometry in chloride and sulfate media as a function of pH in the range 1.5–13. Compounds of Tc(III) and Tc(IV) are produced by reduction of pertechnetate, and the system Tc(III)/Tc(IV) was investigated in acidic media. The potential—acidity diagram of technetium is described for two total pertechnetate concentrations. Evidence for the dismutation of Tc(III) below pH 4 is discussed.     

Electrochemical properties of niosomes modified Au electrode and DNA recognition

Non-ionic surfactant vesicles (NSVs), also referred to as niosomes, have been studied as an alternative to conventional liposomes. In this paper, electrochemical inspection of the interaction between Herring sperm DNA and niosomes has been investigated after a simple and novel method for the formation of niosomes on Au electrode. Each step of electrode modification has been confirmed with cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The assembly of octadecanethiol (ODT) layer on the electrode surface generates a packed film that introduces a barrier to the interfacial electron transfer (R(et)), and the subsequent immobilization of niosomes onto the self-assembled monolayer (SAM) layer results in a further increase of R(et), due to the formed bilayer almost blocked the redox probe to the electrode surface. When Herring sperm DNA was added, the R(et) value decreased, indicating that the barrier of the redox probe to the surface was disrupted. The addition of DNA caused the formation of some transmembrane channels for the redox probe across the niosomes. A good linear relationship between R(et) value and DNA concentration was found over the 0-0.05 mg mL(-1) concentration range.     

Electrochemical properties of niosomes modified Au electrode and DNA recognition

Non-ionic surfactant vesicles (NSVs), also referred to as niosomes, have been studied as an alternative to conventional liposomes. In this paper, electrochemical inspection of the interaction between Herring sperm DNA and niosomes has been investigated after a simple and novel method for the formation of niosomes on Au electrode. Each step of electrode modification has been confirmed with cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The assembly of octadecanethiol (ODT) layer on the electrode surface generates a packed film that introduces a barrier to the interfacial electron transfer (R_et), and the subsequent immobilization of niosomes onto the self-assembled monolayer (SAM) layer results in a further increase of R_et, due to the formed bilayer almost blocked the redox probe to the electrode surface. When Herring sperm DNA was added, the R_et value decreased, indicating that the barrier of the redox probe to the surface was disrupted. The addition of DNA caused the formation of some transmembrane channels for the redox probe across the niosomes. A good linear relationship between R_et value and DNA concentration was found over the 0–0.05 mg mL⁻¹ concentration range.     

AFM studies of solid-supported lipid bilayers formed at a Au(111) electrode surface using vesicle fusion and a combination of Langmuir-Blodgett and Langmuir-Schaefer techniques

Atomic force microscopy (AFM) has been used to characterize the formation of a phospholipid bilayer composed of 1,2-dimyristyl-sn-glycero-3-phosphocholine (DMPC) at a Au(111) electrode surface. The bilayer was formed by one of two methods: fusion of lamellar vesicles or by the combination of Langmuir-Blodgett (LB) and Langmuir-Schaefer (LS) deposition. Results indicate that phospholipid vesicles rapidly adsorb and fuse to form a film at the electrode surface. The resulting film undergoes a very slow structural transformation until a characteristic corrugated phase is formed. Force-distance curve measurements reveal that the thickness of the corrugated phase is consistent with the thickness of a bilayer lipid membrane. The formation of the corrugated phase may be explained by considering the elastic properties of the film and taking into account spontaneous curvature induced by the asymmetric environment of the bilayer, in which one side faces the gold substrate and the other side faces the solution. The effect of temperature and electrode potential on the stability of the corrugated phase has also been described.     

Modulating membrane properties: the effect of trehalose and cholesterol on a phospholipid bilayer

The protective properties of trehalose on cholesterol-containing lipid dipalmitoylphosphatidylcholine (DPPC) bilayers are studied through molecular simulations. The ability of the disaccharide to interact with the phospholipid headgroups and stabilize the membrane persists even at high cholesterol concentrations and restricts some of the changes to the structure that would otherwise be imposed by cholesterol molecules. Predictions of bilayer properties such as area per lipid, tail ordering, and chain conformation support the notion that the disaccharide decreases the main melting transition in these multicomponent model membranes, which correspond more closely to common biological systems than pure bilayers. Molecular simulations indicate that the membrane dynamics are slowed considerably by the presence of trehalose, indicating that high sugar concentrations would serve to avert possible phase separations that could arise in mixed phospholipid systems. Various time correlation functions suggest that the character of the modifications in lipid dynamics induced by trehalose and cholesterol is different in the hydrophilic and hydrophobic regions of the membrane.     

Quantitative SNIFTIRS studies of (bi)sulfate adsorption at the Pt(111) electrode surface

Subtractively normalized interfacial Fourier transform infrared reflection spectroscopy (SNIFTIRS) was applied to study (bi)sulfate adsorption on a Pt(111) surface in solutions of variable pH while maintaining a constant total bisulfate/sulfate ((bi)sulfate) concentration without the addition of an inert supporting electrolyte. The spectra were recorded for both the p- and s-polarizations of the IR radiation in order to differentiate between the IR bands of the (bi)sulfate species adsorbed on the electrode surface from those species located in the thin layer of electrolyte. The spectra recorded with p-polarized light consist of the IR bands from both the species adsorbed at the electrode surface and those present in the thin layer of electrolyte between the electrode surface and ZnSe window whereas the s-polarized spectra contain only the IR bands from the species located in the thin layer of electrolyte. A new procedure was developed to calculate the angle of incidence and thickness of the electrolyte between the Pt(111) electrode surface and the ZnSe IR transparent window. By combining these values with the knowledge of the optical constants for Pt, H(2)O and ZnSe, the mean square electric field strength (MSEFS) at the Pt(111) electrode surface and for thin layer of solution were accurately calculated. The spectra recorded using s-polarization were multiplied by the ratio of the average MSEFS for p- and s-polarizations and subtracted from the spectra recorded using p-polarization in order to remove the IR bands that arise from the species present within the thin layer cavity. In this manner, the resulting IR spectra contain only the IR bands for the anions adsorbed on the Pt(111) electrode surface. The spectra of adsorbed anions show little change with respect to the pH ranging from 1 to 5.6. This behavior indicates that the same species is predominantly adsorbed on the metal surface for this broad range of pH values and the results suggest that sulfate is the most likely candidate for this adsorbate.