F-肌动蛋白与负电荷磷脂膜的相互作用研究

细胞膜的内膜含有大量的负电荷磷脂,研究F-肌动蛋白与负电荷磷脂的相互作用将有助于更深入地了解细胞骨架与细胞膜的体内相互作用机制.在金片和金电极上分别构建了负电荷磷脂的杂化双层磷脂膜,通过表面等离子体共振方法(SPR)和电化学阻抗技术研究了F-肌动蛋白与负电荷磷脂膜的相互作用.结果表明,F-肌动蛋白可以在没有中间联系蛋白的情况下,直接与负电荷磷脂膜发生相互作用.钙离子可以有效地促进它们的相互作用,表明钙离子在其中发挥了重要作用.高浓度的KCl显著抑制它们的相互作用,表明这种相互作用主要受静电作用影响.实验结果进一步证明在F-肌动蛋白与负电荷磷脂膜相互作用时,除了可以通过其它蛋白发生间接相互作用外,还可以与磷脂膜发生直接的相互作用.     

毒素蛋白ColicinE1与磷脂膜相互作用的荧光光谱研究临界摩尔比的测定

磷脂－蛋白相互作用的临界摩尔比是研究膜脂－蛋白相互作用的重要参数．本文利用荧光光谱技术首次测定了毒素蛋白ＣｏｌｉｃｉｎＥ１在不同条件下与不同磷脂膜相互作用的临界摩尔比并通过临界摩尔比的变化讨论了插膜蛋白与磷脂膜相互作用的规律，为进一步探讨毒素蛋白的插膜机制提供了重要的基础     

磷脂膜色谱及其在药物跨膜转运评价中的应用

磷脂膜色谱是固态基质上的有序磷脂分子单层体系采用色谱学方法仿真药物与细胞膜相互作用过程,可用来评价药物的细胞膜渗透性和活性。硅胶表面上的磷脂单分子层模拟了单层细胞膜,因此药物的磷脂膜色谱保留行为可用于预测药物与细胞膜的相互作用。目前考察药物跨膜转运的模型主要有正辛醇/水系统、脂质体/水系统、反相色谱（ODS）以及磷脂膜色谱。与前述3种系统比较,磷脂膜色谱除了具有高效、简便等特点外,同时能模拟药物与生物膜之间疏水作用力以外的其他作用力,因此对磷脂膜色谱的研究也越来越深入。由于药物细胞膜渗透性对其有效性和安全性起着关键作用,因此磷脂膜色谱在新药研发早期阶段的介入可以有效地降低后期候选药物的淘汰率,提高新药的研发效率。该文就磷脂膜色谱的研究及在药物跨膜转运评价中的应用进行了综述。     

毒素蛋白Colicin E1与磷脂膜相互作用的荧光光谱研究——临界摩尔…

磷脂－蛋白相互作用的临界摩尔比是研究膜脂－蛋白相互作用的重要参数。本文利用荧光光谱技术首次测定了毒素蛋白Ｃｏｌｉｃｉｎ Ｅ１在不同条件下与不同磷脂膜相互作用的临界摩尔比并通过临界摩尔比的变化讨论了插膜蛋白与磷脂膜相互作用的规律，为进一步探讨毒素蛋白的插膜机制提供了重要的基础。     

平板双分子层脂膜的制备及锌离子跨该膜的输运过程

制备了氧化胆固醇 卵磷脂(脑磷脂)平板双分子层脂膜,研究了膜配方对双分子层脂膜的稳定性和离子通透性的影响,得到了最佳制膜工艺,建立了锌离子跨卵 (脑 )磷脂膜的吸附 -扩散模型,其计算值与实验值基本吻合.     

磷脂膜的相行为对氧化石墨烯与磷脂膜相互作用的影响

通过使用不同相变温度的磷脂分子并调节二者的比例构筑了不同相态的磷脂膜, 并利用表面增强红外光谱和激光共聚焦显微镜研究了磷脂膜的相行为对氧化石墨烯和磷脂膜相互作用的影响. 结果表明, 氧化石墨烯对磷脂膜中磷脂分子的抽提作用具有显著的相态选择性, 其选择性地抽提流动相的磷脂分子; 氧化石墨烯对流动相磷脂的抽提作用受到膜中凝胶相磷脂存在比例的影响, 只有在流动相磷脂分子占磷脂膜中磷脂分子的绝大部分时才能够发生抽提作用, 且只有流动相的磷脂分子被抽提.     

表面修饰富勒烯的聚酰胺-胺树枝状分子与生物膜的相互作用研究

采用粗粒化的分子动力学模拟,研究了末端修饰富勒烯(C60)的聚酰胺-胺(PAMAM)树枝状分子在水溶液中的结构变化以及其与磷脂膜相互作用行为.发现修饰后的PAMAM分子大小随嫁接数目的增多而减小,而形状变化并不明显,分子仍是近球形结构;同时发现C60分子因疏水作用聚集在一起.与膜作用后,因C60分子插入到双层膜内,导致分子由球形稍微向椭球形改变,分子大小也稍微增大.同时发现PAMAM分子紧紧分布在磷脂膜表面,局域膜厚度变小.     

石墨烯材料与模拟生物膜界面相互作用的研究进展

石墨烯材料在生物领域的蓬勃发展使其纳米生物界面研究已成为纳米生物学研究的热点方向。生物膜是石墨烯材料进入生物体系环境中的第一道屏障,深入理解石墨烯材料与磷脂膜间的相互作用,对于石墨烯基生物材料的功能界面优化设计和生物学效应控制具有极为重要的意义。本文对石墨烯材料进行了简要介绍,系统总结了近几年石墨烯材料与模拟生物膜相互作用的研究进展,并对其研究方向进行了展望。     

大分子拥挤环境中脂肪酸影响磷脂囊泡相变的差示扫描量热研究

脂肪酸诱导的磷脂膜的热力学行为对于认识细胞内复杂的机制有着重要意义,而前人在研究脂肪酸与磷脂膜相互作用时大都在稀溶液中进行;拥挤环境下脂肪酸诱导磷脂膜的相变行为还未见报道。本文以二肉豆蔻酰磷脂酰胆碱(DMPC)构建囊泡模型,采用差示扫描量热法系统地研究了在不同浓度、不同分子量的聚乙二醇(PEG)拥挤环境中不同结构的脂肪酸对DMPC磷脂囊泡相变的影响。研究结果表明,在拥挤环境中,PEG对纯的磷脂囊泡相变的影响与大分子的分子量和浓度相关。对于脂肪酸/磷脂囊泡(FA/DMPC),PEG的存在对囊泡相变产生显著影响。在所考察的分子量和浓度范围内,PEG使FA/DMPC囊泡相变增加。短链饱和脂肪酸、不饱和脂肪酸原本使DPMC囊泡相变降低,但PEG缩小了降低幅度,甚至导致相变增加。进一步的研究表明,在大多数情况下,PEG对FA/DMPC的相变具有协作增强效应,且其影响均与大分子的分子量和浓度相关。另外,随着PEG浓度的升高,磷脂囊泡的协同单位数逐渐降低,表明拥挤环境会影响磷脂双分子层的均一性,使协同发生相变的分子数降低。本文的研究表明,大分子拥挤环境能够对扰动的磷脂双分子层起到一定的修复作用,这一现象在生物膜相关领域不可忽视。     

锆镁磷脂膜色谱固定相的制备及其在评价药物-膜相互作用中的应用

基于锆基质与磷脂之间强烈的路易斯酸碱作用,制备了锆镁磷脂膜色谱固定相,并使用红外光谱、X射线光电子能谱对该色谱固定相进行了表征;使用与体内环境类似的生理缓冲液体系为流动相,评价了该模拟生物膜色谱固定相预测药物膜渗透性的能力,结果表明药物在锆镁磷脂膜色谱中的保留(log Kmbm)与表观渗透率(log Papp)在预测药物的膜渗透性、跨膜吸收等方面具有非常好的相关性,相关系数为0.970,斜率接近1。通过理论推导,引入了直观、方便的热力学指标吉布斯自由能差值（Δ(ΔG°)）对药物-膜之间的相互作用强弱进行了评价。     

Au/硫醇/卵磷脂双层膜的制备及其电化学表征

生命体系的许多过程发生在生物膜上,生物膜模拟体系的制备与研究一直受到人们的重视．自组装单分子膜（ＳｅｌｆＡｓｅｍｂｌｅｄＭｏｎｏｌａｙｅｒｓ,ＳＡＭｓ）特别是在金基底表面的硫醇单分子膜由于其具有良好的稳定性及有序性,因此在基础理论研究及应用技术等方...     

Adiabatic compressibility of red blood cell membrane: influence of skeleton

Measurements of ultrasound velocity and density were used for determination of the adiabatic compressibility of red blood cells (RBC) during detachment of the membrane skeleton. Skeleton detachment was induced by addition of nystatin into a low ionic strength RBC suspension resulting in an increase (10%) of the ultrasound velocity concentration increment, [u], while the specific volume of cells, phi(v) did not change significantly. Changes of the concentration increment had rather long kinetics and were not completed even after 60 min. Both [u] and phiV values were used for calculation of the specific apparent adiabatic compressibility of RBC, phiK/beta0. The value of the specific apparent compressibility decreases following addition of nystatin. This corresponds to an increase in the volume elastic modulus of RBC membranes during detachment of the membrane skeleton. Control experiments with large unilamellar liposomes at conditions similar to that performed with the RBC did not reveal significant changes of [u] after the addition of nystatin. Our results show that the role of the membrane skeleton probably consists in maintaining higher compressibility of the RBC membranes. This may partly provide conditions for conformational changes of RBC membrane proteins.     

Simultaneous determination of amounts of major phospholipid classes and their fatty acid composition in erythrocyte membranes using high-performance liquid chromatography and gas chromatography.

A method for the simultaneous determination of amounts of major phospholipid classes and their fatty acid composition in erythrocyte membranes is described. The method consists in extraction of phospholipids from erythrocyte membranes, separation of phospholipid classes by high-performance liquid chromatography, methylation of phospholipids and determination of phospholipid-bound fatty acids by capillary gas chromatography. The amounts of phospholipid classes are calculated from the total weight of phospholipid-bound fatty acids and their average molecular weights. The method was applied to erythrocytes from rats. The results show that the method is reproducible and is useful for the determination of amounts of phospholipid classes and their fatty acid composition in small blood samples.     

HYDROPHOBIZING EFFECT OF CATIONS ON ACIDIC PHOSPHOLIPID MEMBRANES

By means of contact angle measurements with water and aqueous salt solutions, it is shown that plurivalent cations increase the hydrophobicity of negatively charged phospholipid vesicle membranes (consisting of phosphatidic acid, PA, or of phosphatidylserine, PS), but does not influence the hydrophobicity of neutral phospholipid membranes, (e.g., phosphatidylcholine, PC, at up to 200 mM of CaCl₂). The hydrophobizing action of cations on PA and PS membranes is concomitant with the reduction in (negative) zeta potential with increasing cation concentrations. Trivalent cations, La³⁺, showed more effective in hydrophobizing negatively charged phospholipid membranes than divalent and monovalent cations. Except for hydrogen ions, monovalent cations do not show any appreciable hydrophobizing effect on lipid vesicle membranes at concentrations less than 1 M. The hydrophobizing effect on phospholipid membranes can also be used to explain the induction of lateral phase separation into patches of different phospholipids as well as cell fusion.     

Aliphatic polyurethane as a matrix for pH sensors: effects of native sites and added proton carrier on electrical and potentiometric properties

The potentiometric and impedance characteristics of polymeric membranes, based on aliphatic polyurethane (Tecoflex) as a matrix, are described and interpreted by theory and experiments for H(+) and alkali metal ion-sensitive sensors. Both dummy plasticized membranes and proton carrier-loaded membranes can show pH response. The pH response of dummy membranes is due to protonated natural negative sites in the polyurethane matrix. The electrodes with added proton carrier show improved rejection of Li(+), Na(+), and K(+) responses and give useful analytical responses. Optimal performance requires control of negative site concentration by addition of lipophilic salt (e.g. tetraphenylborate derivatives). Impedance analyses show surface-rate semicircles and, depending on the bathing electrolyte solution, appearance of a diffusional Warburg impedance. In addition to these time-dependence surface region effects, changes in the bulk membrane resistance with soaking time can be well correlated with equilibrium water content of plasticized membranes.     

Electrochemical impedance spectroscopy fingerprints the ion selectivity of microgel functionalized ion-exchange membranes

Surface modification methods are applied to alter interfacial phenomena and improve ion transport through membranes. In this work we present a novel method for tailoring the surface of cation-exchange membranes based on the deposition of thin microgel monolayers. The charge of such layers exerts a strong influence on the monovalent-ion-selectivity, and this is reflected in the electrochemical impedance responses. Membranes coated with uncharged microgels show similar behavior to that of unmodified ones, with impedance spectra dominated by low-frequency diffusional arcs. However, membranes modified with positively charged microgels exhibit an increased resistance due to the hindered transport of cations through the modification. An additional high-frequency capacitive arc is obtained with the monovalent-ion-selective membranes, which is attributed to concentration polarization effects at the membrane/modification interface. The characteristic frequency of this arc decreases with the valency of the ion, thus proving that multivalent ions pass through the modification layer at rates much slower than monovalent ones. Accordingly, electrochemical impedance spectroscopy has been used to feature monovalent-ion-selective properties of layered membranes.     

Assessing the Apparent Effective Thickness of the Supported Hybrid Bilayer Membranes Consisting of Octadecanethiol and Phospholipid by ac Impedance Spectroscopy

The electrochemical behavior of the gold electrode modified by hybrid bilayer membranes in different concentrations of Fe(CN)₆^3?/Fe(CN)₆^4? was investigated by electrochemical impedance spectroscopy and cyclic voltammetry technology. The electron transfer between gold and the redox species separated by the hybrid bilayers assembly was completed by an electron tunneling process. A detailed equivalent circuit for electron transport across the HBMs is proposed. It was found that the apparent effective thickness of the hybrid bilayer membranes was lower than that of the sum of the chain length of octadecanethiol and phospholipid so some possible collapsed sites might exist at the hybrid bilayer membranes.     

Detecting cross talk between two halves of a phospholipid bilayer

One of the most challenging questions that relates to the structure and function of biological membranes is whether the two halves of the bilayer "talk" to each other. In this letter, we show how the perturbation of the lateral organization of one leaflet of a fluid phospholipid bilayer by an external agent also alters the lateral organization of the adjoining leaflet. In addition, we show that the energy involved in such "cross talk" corresponds to ca. 100 cal/mol of phospholipid. These findings provide a basis for expecting similar cross talk to exist in cell membranes.     

Bacteriorhodopsin-containing membrane fragments adsorbed on mercury-supported biomimetic membranes

Membrane fragments containing bacteriorhodopsin (bR), a light-driven proton pump, are adsorbed on mercury-supported biomimetic membranes. The biomimetic membranes consist in a phospholipid monolayer and an alkanethiol/phospholipid bilayer supported by a hanging mercury drop electrode. Once adsorbed on the lipid film, bR is activated by shining the mercury drop with an appropriate light source, and the transient capacitive current generated by the ion pump is measured under short-circuit conditions and in the absence of photoartefacts. Our results show that the lipid-coated mercury electrode is suitable for adsorbing membrane fragments and, hence, for studying the electrochemical behaviour of protein in a biomimetic environment.     

Electrochemical impedance spectroscopy of tethered bilayer membranes

The electrochemical impedance spectra (EIS) of tethered bilayer membranes (tBLMs) were analyzed, and the analytical solution for the spectral response of membranes containing natural or artificially introduced defects was derived. The analysis carried out in this work shows that the EIS features of an individual membrane defect cannot be modeled by conventional electrical elements. The primary reason for this is the complex nature of impedance of the submembrane ionic reservoir separating the phospholipid layer and the solid support. We demonstrate that its EIS response, in the case of radially symmetric defects, is described by the Hankel functions of a complex variable. Therefore, neither the impedance of the submembrane reservoir nor the total impedance of tBLMs can be modeled using the conventional elements of the equivalent electrical circuits of interfaces. There are, however, some limiting cases in which the complexity of the EIS response of the submembrane space reduces. In the high frequency limit, the EIS response of a submembrane space that surrounds the defect transforms into a response of a constant phase element (CPE) with the exponent (α) value of 0.5. The onset of this transformation is, beside other parameters, dependent on the defect size. Large-sized defects push the frequency limit lower, therefore, the EIS spectra exhibiting CPE behavior with α ≈ 0.5, can serve as a diagnostic criterion for the presence of such defects. In the low frequency limit, the response is dependent on the density of the defects, and it transforms into the capacitive impedance if the area occupied by a defect is finite. The higher the defect density, the higher the frequency edge at which the onset of the capacitive behavior is observed. Consequently, the presented analysis provides practical tools to evaluate the defect density in tBLMs, which could be utilized in tBLM-based biosensor applications. Alternatively, if the parameters of the defects, e.g., ion channels, such as the diameter and the conductance are known, the EIS data analysis provides a possibility to estimate other physical parameters of the system, such as thickness of the submembrane reservoir and its conductance. Finally, current analysis demonstrates a possibility to discriminate between the situations, in which the membrane defects are evenly distributed or clustered on the surface of tBLMs. Such sensitivity of EIS could be used for elucidation of the mechanisms of interaction between the proteins and the membranes.