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。     

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（英文）

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。     

免标记地检测二元磷脂膜中磷脂分布的表面增强拉曼成像方法

以银纳米粒子自组装层为增强基底,我们报道了一种用于检测二元磷脂膜中具有相似结构磷脂分布的表面增强拉曼成像方法,这种方法具有免标记及花费低廉的优点.对探针分子对巯基苯胺（p-aminothiophenol）,实验中所用的银纳米粒子自组装层表现出强的表面增强拉曼活性及良好的重现性.原子力显微镜表征结果证明了完整的磷脂膜在银纳米粒子自组装层上的形成.以这种银自组装层为基底,我们得到了磷脂膜中二肉豆蔻酰磷脂酰甘油（DMPG）和二肉豆蔻酰磷脂酰胆碱（DMPC）的表面增强拉曼光谱,并且利用DMPG的光谱特征峰,1482cm-1,区分这两种磷脂.而通过1482cm-1和1650cm-1的峰强比（R1482/1650）,可以同时得知在混合磷脂膜上某点这两种磷脂所占的比例：R1482/1650值的增加意味着DMPG的增加和DMPC的减少.磷脂膜的表面增强拉曼成像则是由R1482/1650值和对应的位置信息组合而得到,其成像结果表明了带电的磷脂DMPG在混合磷脂膜中的聚集.我们所报道的基于表面增强拉曼成像技术的方法提供了一种便利的、免标记的和花费低廉的途径来研究磷脂膜的结构,例如磷脂域和脂阀.     

可聚合囊泡体系(烯丙基-二甲基-十二烷基溴化胺和十二烷基硫酸钠)在溶液中的盐效应

高稳定的囊泡广泛用于制作生物模型、药物输送以及合成纳米材料的模板。获得高稳定囊泡结构的重要方法之一是用聚合反应固定囊泡结构。作为可聚合囊泡制备的前期基础工作,研究了一种可聚合的囊泡体系：1-丙烯基－2,2,二甲基－十二烷基溴化胺（ADDB）和ADDB与十二烷基磺酸钠（SDS）的等摩尔比混合体系。该囊泡体系即使在高浓度盐水中也能够自发地形成均相的囊泡溶液。在聚合之前,采用动态激光光散射（DLS）、冷冻蚀刻透射电镜（FF-TEM）技术研究了可聚合囊泡的盐效应。DLS测试发现没有盐存在时,囊泡大小为83 nm,盐的浓度增加到250 mmol/L时,囊泡尺寸增大到250 nm。然而继续增大盐浓度到1000 mmol/L, 囊泡尺寸减小到180nm. FF-TEM结果发现盐浓度小于150 mM时, 单个囊泡为70 nm左右,然而明显存在囊泡的絮凝与融合;当盐浓度增加到400 mM时,单个囊泡尺寸减小到20 nm. 因此DLS 观测到囊泡尺寸增大的原因是由于囊泡的絮凝与融合;而尺寸减小的原因是由于在高盐浓度下,盐屏蔽了带电颗粒之间的静电相互作用,在熵增的驱使下,大囊泡变成小囊泡。     

巨型囊泡的侧向相分离与出芽

用电形成法制备含三组分二油酰磷脂酰胆碱(DOPC)/二棕榈酰磷脂酰胆碱(DPPC)/胆固醇(Chol)的巨型囊泡, 以TR-DPPE为荧光染色剂, 在荧光显微镜下直接观察膜的侧向相分离与微区相凸起出芽的耦合. 发现囊泡膜内的相分离具有诱导期, 相分离速度很快, 形成的微相区在整个囊泡球面上均衡分布. 各微相区的出芽不是同时进行, 为逐个随机发生. 每次出芽的时间小于0.5 s. 分相与出芽的耦合使球面上的不同微区之间不会相互融合成更大的微区.     

纳米雄黄与脂质体仿生膜的相互作用研究

本工作以卵磷脂与胆固醇组成的磷脂小单层脂质体（small unilamelarvesicles,suv）作为仿生膜的简单模型,采用表面等离子共振技术（SPR）、荧光偏振、拉曼（Raman）光谱、核磁共振（NMR）及原子力显微镜（AFM）研究纳米雄黄与SUV仿生膜的相互作用,证实了磷脂是纳米雄黄作用的关键靶分子．随纳米雄黄结合,SUV仿生膜的相对粘度聃值增大,膜的流动性减小．Raman光谱数据计算表明,作用后膜的纵向有序性参数s。。及横向有序性参数Slat值增大,说明纳米雄黄的结合使磷脂膜的脂酰基链全反式构型比例上升,膜的流动性减小．由Raman光谱和引PNMR结果推测,磷脂极性头部是纳米雄黄与磷脂的主要结合位点。AFM实时观测,纳米雄黄通过在膜表面打“孔”或“洞”的方式,损坏磷脂膜．     

支撑磷脂双层膜的研究进展

支撑磷脂双层膜(supported phospholipid bilayers,SPBs)是细胞膜研究中普及的模型,是固定生物活性物质的理想材料,不仅可以保持生物分子的活性,还能有效抑制其他生物分子的非特异性吸附,在跨膜蛋白、仿生膜、水处理、生物医学和生物传感器等研究领域具有广泛的应用前景。本文介绍了支撑磷脂双层膜的表征方法和制备方法,包括Langmuir Blodgett(LB)膜提拉法、囊泡融合法和LB膜提拉法与囊泡融合联合法;详细阐述了囊泡融合法制备SPBs的机理;综述了囊泡融合法制备SPBs的影响因素,包括囊泡浓度、缓冲溶液、温度、囊泡和基底表面电荷等因素;列举了支撑磷脂膜的应用,并展望了支撑磷脂双层膜的研究趋势。     

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.     

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。     

pH对磷脂膜中Slc11a1跨膜区二级结构和取向的影响

采用圆二色光谱、 荧光光谱、 红外衰减全反射光谱和差示扫描量热分析等方法对不同pH条件下膜蛋白Slc11a1(溶质转运蛋白家族11成员1)的第二、 第三和第四跨膜区(TMD2~TMD4)在磷脂膜[二肉豆蔻酰磷脂酰胆碱(DMPC)和二肉豆蔻酰磷脂酰甘油(DMPG)的摩尔比为2∶1]中的二级结构和取向进行了研究. 结果表明, TMD3的二级结构及在磷脂膜内的位置与pH密切相关, 在pH=7时TMD3主要为β股结构, 在膜中埋入较浅; 而在pH=5.5时TMD3形成部分α螺旋结构, 并较深地埋入膜中. 对TMD3进行E139A突变后的结果证明, TMD3的这些性质与位于中间的谷氨酸的质子化性质密切相关. 实验结果还表明, TMD2和TMD4在不同pH条件下都形成α螺旋结构并分别以26°和35°的倾斜角插入磷脂膜内, 它们在磷脂膜内的位置基本不受pH影响.     

静电聚结技术在油水分离中的研究及应用进展

静电聚结技术具有快速、清洁、高效的特性,通常不需要添加化学药剂,不产生附加污染物,同重力沉降等方法相比,对于小粒径水滴或油水界面稳定的油水混合液适应性更强。本文简要介绍了静电聚结技术的聚结原理,总结了传统电脱水技术、管式静电预聚结技术和容器内置式静电聚结技术(VIEC)的研究和应用进展,对比分析了三类静电聚结技术的适用领域及技术特点,介绍了静电聚结与湍流/剪切流耦合技术的研究现状,最后对静电聚结技术的发展方向进行归纳和展望。     

The Role of Molecular Electrostatic Potentials in the Formation of a Halogen Bond in Furan⋅⋅⋅XY and Thiophene⋅⋅⋅XY Complexes

The halogen bonding of furan???XY and thiophene???XY (X=Cl, Br; Y=F, Cl, Br), involving σ‐ and π‐type interactions, was studied by using MP2 calculations and quantum theory of “atoms in molecules” (QTAIM) studies. The negative electrostatic potentials of furan and thiophene, as well as the most positive electrostatic potential (V_S,max) on the surface of the interacting X atom determined the geometries of the complexes. Linear relationships were found between interaction energy and V_S,max of the X atom, indicating that electrostatic interactions play an important role in these halogen‐bonding interactions. The halogen‐bonding interactions in furan???XY and thiophene???XY are weak, “closed‐shell” noncovalent interactions. The linear relationship of topological properties, energy properties, and the integration of interatomic surfaces versus V_S,max of atom X demonstrate the importance of the positive σ hole, as reflected by the computed V_S,max of atom X, in determining the topological properties of the halogen bonds.     

Topography of approximate molecular electrostatic potentials

A new method is described for the approximation of the molecular electrostatic potential (MESP). This method is used for the study of the topography of small molecules. The critical points of the approximate and the exact MESP are compared. It is found that most of the critical points of the exact MESP are retained, but in regions where the exact MESP changes slowly near critical points the number of critical points of the approximate MESP can be reduced.     

Steric vs. electrostatic effects in the nucleophilic addition to a hindered cyclohexanone

The relative importance of steric vs. electrostatic effects in the nucleophilic addition to (4R,6S)-4-(tert-butyldimethylsiloxy)-2,2,6-trimethylcyclohexanone (1), a well-known chiral building block, is investigated.     

Theoretical studies of the mechanism of the action of the neurohypophyseal hormones. I. Molecular electrostatic potential (MEP) and molecular electrostatic field (MEF) maps of some vasopressin analogues

Summary Continuing our theoretical studies of the oxytocin and vasopressin analogues, we have analysed the molecular electrostatic potential (MEP) and the norm of the molecular electrostatic field (MEF) of [1--mercaptopropionic acid]-arginine-vasopressin ([Mpa¹]-AVP), [1-(-mercapto-,-cyclopentamethylene)propionic acid]-arginine-vasopressin ([Cpp]-AVP), and [1-thiosalicylic acid]-arginine-vasopressin ([Ths]-AVP) whose low-energy conformations were calculated in our previous work. These compounds are known from experiment to exhibit different biological activity. The scalar fields mentioned determine the energy of interaction with either charged (MEP) or polar (MEF) species, the energy being in the second case either optimal or Boltzmann-averaged over all the possible orientations of the dipole moment versus the electrostatic field. The electrostatic interactions slowly vanish with distance and can therefore be considered to be the factor determining the molecular shape at greater distances, which can help in both predicting the interactions with the receptor at the stage of remote recognition and in finding the preferred directions of solvation by a polar solvent. In the analysis of the fields three techniques have been used: (i) the construction of maps in certain planes; (ii) the construction of maps on spheres centered in the charge center of the molecule under study and of poles chosen according to the main axes of the quadrupole moment; and (iii) the construction of surfaces corresponding to a given value of potential. The results obtained show that the shapes of both MEP and MEF are similar in the case of [Mpa¹]-AVP and [Cpp¹-AVP (biologically active), while some differences emerge when comparing these compounds with [Ths¹]-AVP (inactive). It has also been found that both MEP and MEF depend even more strongly on conformation.     

Influence of polarization functions on molecular electrostatic potentials

We study the effects of d-polarization functions, centered on the heavy atoms, on the SCF molecular electrostatic potentials calculated for some molecules. The positions and energies of the minima found are very sensitive to the inclusion of polarization functions in the basis set used. The variations depend on the heavy atom involved and on the possible anisotropy of its charge distribution. These variations are particularly important for second-row atoms.     

The Poisson-Boltzmann model for tRNA: Assessment of the calculation set-up and ionic concentration cutoff

Using tRNA molecule as an example, we evaluate the applicability of the Poisson-Boltzmann model to highly charged systems such as nucleic acids. Particularly, we describe the effect of explicit crystallographic divalent ions and water molecules, ionic strength of the solvent, and the linear approximation to the Poisson-Boltzmann equation on the electrostatic potential and electrostatic free energy. We calculate and compare typical similarity indices and measures, such as Hodgkin index and root mean square deviation. Finally, we introduce a modification to the nonlinear Poisson-Boltzmann equation, which accounts in a simple way for the finite size of mobile ions, by applying a cutoff in the concentration formula for ionic distribution at regions of high electrostatic potentials. We test the influence of this ionic concentration cutoff on the electrostatic properties of tRNA.     

Nonbonding pairs in cyclic thioethers: Electrostatic modeling and ab initio calculations for complexes of 2,5-dihydrothiophene,thietane, and thiirane with hydrogen fluoride

Electrostatic potential energies V(ϕ) of a non-perturbing, protonic charge at fixed distances r from the S atom in three cyclic thioethers were examined as functions of the angles ϕ made by the r-vector with the C₂ axis (thiirane and 2,5-dihydrothiophene) or the local C₂ axis (thietane). The electrostatic PE V_HF(ϕ) of HF (HF modelled as an extended electric dipole) was also calculated and the results compared with geometries of the thioether⋯HF complexes calculated at the CCSD(T)-F12c/cc-pVTZ-F12 level. The latter reveal angular deviations θ ∼10-20° of the S⋯H F nuclei from collinearity in directions suggesting secondary interactions of F with H atom(s) of the rings. Angles ϕ made by the S⋯H hydrogen bond with the C₂ (or local C₂) axes in the complexes are systematically larger (∼4-9°) than indicated by the V_HF(ϕ) functions. Minima in the simple V(ϕ) versus ϕ functions occur at values smaller (∼5-10°) than those in the V_HF(ϕ) curves.     

Electrostatic potentials and fields from density expansions of deformed atoms in molecules

The exact representation of the molecular density by means of atomic expansions, consisting in spherical harmonics times analytical radial factors, is employed for the calculation of electrostatic potentials, fields, and forces. The resulting procedure is equivalent to an atomic multipolar expansion in the long-range regions, but works with similar efficiency and accuracy in the short-range region, where multipolar expansions are not valid. The performances of this procedure are tested on the calculation of the electrostatic potential contour maps and electrostatic field flux lines of water and nitrobenzene, computed from high-quality molecular electron densities obtained with Slater basis sets.     

CoMFA Analysis of Biodegradability

Abstract

CoMFA (comparative molecular field analysis) has been used to correlate the biodegradability of several classes of compounds. The technique uses an atomic probe to detect the steric and electrostatic fields around a molecule. Good cross-validated correlations were obtained for some series (alcohols, carboxylic acids and linear alkyl benzene sulphonates). Correlations were weaker for esters and benzene sulphonates, and were non-existent for phenols. These results may reflect the unreliability of biodegradation data, but may also be a result of molecular misalignment in the CoMFA procedure. CoMFA appears to be a potentially very useful method for the prediction of biodegradability.