Trichosanthin can spontaneously penetrate phospholipid monolayer under acid condition

Tianhuafen is a kind of traditional Chinese medicine for abortion, which has long been used in China. The basic protein trichosanthin (TCS) is responsible for such activity. As a ribosome inactivating protein (RIP), trichosanthin removes A4304 in the 28s rRNA via the N-glycosidase activity and inactivates the ribosome. However, it remains unclear how TCS can enter the cytoplasm. In the experiment, monolayers at air/water interface have been used to study the interaction between TCS and phospholipid membrane. The results show that TCS can penetrate negatively charged phospholipid monolayer under acid condition, and its membrane penetrating ability is obviously pH-dependent. A hypothesis of TCS penetrating biological membrane under acid condition is proposed based on the obtained result.     

Interaction of SynaptotagminⅠ with Phospholipid Membrane: A Monolayer Study

IntroductionSynaptotagmin is a family of vesicletransmembrane proteins present in synapticvesicles and large secretary granules of neuronsand endocrine cells[1 ] .It is a major constituent ofsynaptic vesicle membranes,comprising7% 8%of the total vesicle protein,characterized by ashort intravesiclar N-terminus,a singletransmembrane region,and a long plasmicdomain.　The best-charaterized form of synaptotagmin,syt ,is found abundantly in rostrol brain.Syt was first described in 1 981 [2 ] ,and i…     

蛋氨酸在长链硫醇修饰电极上的电化学行为研究

研究了蛋氨酸在ＨＳ（ＣＨ）１１ＮＨ２,ＨＳ（ＣＨ２）１１ＣＯＯＨ,ＨＳ（ＣＨ２）１１ＯＨ,ＨＳ（ＣＨ２）１１ＣＨ３４活跃步同末端官能团长链硫醇修饰电极上的电化学行为。结果表明ＤＬ－Ｍｅｔ不仅可以渗透自组单分子层膜,而且ＤＬ－Ｍｅｔ在不同末端官能团的ＳＡＭｓ上的渗秀速率不同。     

Phospholipids and the origin of cationic gating charges in voltage sensors

Cells communicate with their external environment through physical and chemical processes that take place in the cell-surrounding membrane. The membrane serves as a barrier as well as a special environment in which membrane proteins are able to carry out important processes. Certain membrane proteins have the ability to detect the membrane voltage and regulate ion conduction or enzyme activity. Such voltage-dependent processes rely on the action of protein domains known as voltage sensors, which are embedded inside the cell membrane and contain an excess of positively charged amino acids, which react to an electric field. How does the membrane create an environment suitable for voltage sensors? Here we show under a variety of conditions that the function of a voltage-dependent K+ channel is dependent on the negatively charged phosphodiester of phospholipid molecules. A non-voltage-dependent K+ channel does not exhibit the same dependence. The data lead us to propose that the phospholipid membrane, by providing stabilizing interactions between positively charged voltage-sensor arginine residues and negatively charged lipid phosphodiester groups, provides an appropriate environment for the energetic stability and operation of the voltage-sensing machinery. We suggest that the usage of arginine residues in voltage sensors is an adaptation to the phospholipid composition of cell membranes.     

四乙基溴化铵对离子型表面活性剂胶束化行为的影响

用表面张力方法研究了有机电解质四乙基溴化铵（（Ｅｔ）４ＮＢ）对离子型表面活性剂ＳＤＳ和ＤＴＡＢ胶束化行为的影响,并与无机电解质ＮａＢｒ的结果比较．实验结果表明,（Ｅｔ）４ＮＢ促进ＳＤＳ胶束化的能力强于ＮａＢｒ;但促进ＤＴＡＢ胶束化的能力与ＮａＢｒ基本一样．这说明：（Ｅｔ）４Ｎ＋与ＳＤＳ胶束间除了相反电荷静电吸引外,其乙基和表面活性剂碳氢链间还具有弱疏水相互作用,两者结合使（Ｅｔ）４Ｎ＋嵌入ＳＤＳ胶束离子头基中;这种疏水作用强度不及静电力,在静电排斥下,（Ｅｔ）４Ｎ＋乙基和ＤＴＡ＋碳氢链间疏水相互作用未能发生．     

Structure of the C2 domain of human factor VIII at 1.5 A resolution

Human factor VIII is a plasma glycoprotein that has a critical role in blood coagulation. Factor VIII circulates as a complex with von Willebrand factor. After cleavage by thrombin, factor VIIIa associates with factor IXa at the surface of activated platelets or endothelial cells. This complex activates factor X (refs 6, 7), which in turn converts prothrombin to thrombin in the presence of factor Va (refs 8, 9). The carboxyl-terminal C2 domain of factor VIII contains sites that are essential for its binding to von Willebrand factor and to negatively charged phospholipid surfaces. Here we report the structure of human factor VIII C2 domain at 1.5 A resolution. The structure reveals a beta-sandwich core, from which two beta-turns and a loop display a group of solvent-exposed hydrophobic residues. Behind the hydrophobic surface lies a ring of positively charged residues. This motif suggests a mechanism for membrane binding involving both hydrophobic and electrostatic interactions. The structure explains, in part, mutations in the C2 region of factor VIII that lead to bleeding disorders in haemophilia A.     

离子型磁性液体中纳米微粒间相互作用研究

用磁偶极子硬球模型对磁性液体中磁性纳米微粒间相互作用进行了研究。分析表明,微粒间存在着静电排斥、静磁吸引和Van der Waals吸引三种相互作用,无外场时静磁作用非常微弱,可以忽略,静电排斥力作用距离远,起抗团聚作用,Van der Waals作用距离短,是引起团聚主要原因。为确保磁性液体稳定性,制备的离子型磁性液体体积分数应小于5%。     

Removal of phospho-head groups of membrane lipids immobilizes voltage sensors of K+ channels

A fundamental question about the gating mechanism of voltage-activated K+ (Kv) channels is how five positively charged voltage-sensing residues in the fourth transmembrane segment are energetically stabilized, because they operate in a low-dielectric cell membrane. The simplest solution would be to pair them with negative charges. However, too few negatively charged channel residues are positioned for such a role. Recent studies suggest that some of the channel's positively charged residues are exposed to cell membrane phospholipids and interact with their head groups. A key question nevertheless remains: is the phospho-head of membrane lipids necessary for the proper function of the voltage sensor itself? Here we show that a given type of Kv channel may interact with several species of phospholipid and that enzymatic removal of their phospho-head creates an insuperable energy barrier for the positively charged voltage sensor to move through the initial gating step(s), thus immobilizing it, and also raises the energy barrier for the downstream step(s).     

用原子力显微镜和单层膜技术研究短杆菌肽D与卵磷脂混合物的界面性质(英文)

短杆菌肽是一种被广泛研究的细胞膜中的多肽．其与磷脂之间的相互作用以及其在磷脂单层膜中的构象还不十分清楚．研究发现短杆菌肽Ｄ在卵磷脂单层膜中有自发形成聚集体的趋势，而且聚集体的形状依赖于短杆菌肽Ｄ在膜中的浓度．当浓度特别低时，聚集体很大；随着浓度的增加，聚集体逐渐变高但面积逐渐变小．特别是当短杆菌肽Ｄ的摩尔百分率达到１０％时，聚集体几乎形成圆锥型     

羟基取代双烷烃链苯磺酸盐水气界面单层膜的分子动力学模拟

采用分子动力学模拟方法考察2-羟基-3,5-烷基苯磺酸钠表面活性剂分子在水气界面形成单层膜的过程,通过体系各组分的密度分布分析单层膜的形态,采用径向分布函数和氢键分布考察含有羟基的极性基在界面处的吸附构型,并根据转动时间相关函数对双疏水链的吸附构型进行分析。结果表明:羟基的存在减弱了极性基之间的静电排斥作用,增加了分子的饱和吸附量;两条烷烃链具有不同的界面吸附构型,烷烃链间的叠加缠绕增强了溶剂对表面活性剂分子的疏水力作用。     

Negatively and positively charged bacterial aerosol concentration and diversity in natural environments

Bioaerosol charge information is of vital importance for their electrostatic collection. Here, electrostatic means and molecular tools were applied to studying bioaerosol charge dynamics. Positively or negatively charged bioaerosols were collected using an electrostatic sampler operated with a field strength of 1.1 kV cm 1 at a flow rate of 3 L min 1 for 40 min. Those with fewer or no charges bypassing the sampler were also collected using a filter at the downstream of the electrostatic sampler in one environment. The experiments were independently conducted three times in three different environments. The collected bacterial aerosols were cultured directly on agar plates at 26°C, and the colony forming units (CFU) were manually counted. In addition, the CFUs were washed off from the agar plates, and further subjected to polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE) for culturable diversity analysis. The results revealed remarkable differences in positively and negatively charged culturable bacterial aerosol concentration and diversity among the studied environments. In the office environment, negatively charged culturable bacterial aerosols appeared to dominate (P = 0.0489), while in outdoor and hotel environments both polarities had similar concentration levels (P = 0.078, P = 0.88, respectively). DGGE patterns for positively charged culturable bacterial aerosols were shown strikingly different from those of negatively charged regardless of the sampling environments. In addition, for each of the environments positively charged culturable bacterial aerosols collected were found to have more band pattern similarity with those positively charged for respective regions of agar plates than those negatively charged, and vice versa. The information developed here is useful for developing efficient electrostatic sampling protocols for bioaerosols.     

Electric-field-induced capillary attraction between like-charged particles at liquid interfaces

Nanometre- and micrometre-sized charged particles at aqueous interfaces are typically stabilized by a repulsive Coulomb interaction. If one of the phases forming the interface is a nonpolar substance (such as air or oil) that cannot sustain a charge, the particles will exhibit long-ranged dipolar repulsion; if the interface area is confined, mutual repulsion between the particles can induce ordering and even crystallization. However, particle ordering has also been observed in the absence of area confinement, suggesting that like-charged particles at interfaces can also experience attractive interactions. Interface deformations are known to cause capillary forces that attract neighbouring particles to each other, but a satisfying explanation for the origin of such distortions remains outstanding. Here we present quantitative measurements of attractive interactions between colloidal particles at an oil-water interface and show that the attraction can be explained by capillary forces that arise from a distortion of the interface shape that is due to electrostatic stresses caused by the particles' dipolar field. This explanation, which is consistent with all reports on interfacial particle ordering so far, also suggests that the attractive interactions might be controllable: by tuning the polarity of one of the interfacial fluids, it should be possible to adjust the electrostatic stresses of the system and hence the interparticle attractions.     

多巴胺及抗坏血酸在CTAB/CS复合物膜修饰电极上的电化学行为及其测定

制备了十六烷基三甲基溴化铵(CTAB)/壳聚糖(CS)复合物膜修饰电极,研究了多巴胺(DA)和抗坏血酸(AA)在该修饰电极以及裸电极上的电化学行为。循环伏安法研究表明,CTAB与带负电荷AA之间的静电作用使AA的氧化过电位降低,DA和AA两者的峰电位差达360 mV,使得DA和AA重叠氧化峰得以分离。以此建立了同时测定DA和AA的电化学方法。微分脉冲伏安法研究结果表明,DA和AA氧化峰电流和其相应浓度分别在1×10~(-5)～2.8×10~(-3) mol/L和5×10~(-6)～6×10~(-4) mol/L的范围内呈良好的线性关系。应用该方法对DA和AA进行了选择性测定研究,取得良好结果。     

Electrostatic interactions in the assembly of haemoglobin

Haemoglobin is the prototype of an allosteric protein in which cooperative behaviour depends on interaction between unlike subunits. Here we present haematological and biochemical evidence that electrostatic interactions are an important determinant of haemoglobin assembly. Individuals heterozygous for positively charged beta-globin variants have a significantly lower proportion of abnormal haemoglobin than those with negatively charged variants. Moreover, these differences become more pronounced when alpha-thalassaemia is also present. Kinetic experiments using isolated chains indicate that the rate of assembly of the heterotetramer is influenced by alterations in surface charge. A simple electrostatic model is proposed in an attempt to explain these haematological and experimental findings.     

Crystal structures of the membrane-binding C2 domain of human coagulation factor V

Rapid and controlled clot formation is achieved through sequential activation of circulating serine proteinase precursors on phosphatidylserine-rich procoagulant membranes of activated platelets and endothelial cells. The homologous complexes Xase and prothrombinase, each consisting of an active proteinase and a non-enzymatic cofactor, perform critical steps within this coagulation cascade. The activated cofactors VIIIa and Va, highly specific for their cognate proteinases, are each derived from precursors with the same A1-A2-B-A3-C1-C2 architecture. Membrane binding is mediated by the C2 domains of both cofactors. Here we report two crystal structures of the C2 domain of human factor Va. The conserved beta-barrel framework provides a scaffold for three protruding loops, one of which adopts markedly different conformations in the two crystal forms. We propose a mechanism of calcium-independent, stereospecific binding of factors Va and VIIIa to phospholipid membranes, on the basis of (1) immersion of hydrophobic residues at the apices of these loops in the apolar membrane core; (2) specific interactions with phosphatidylserine head groups in the groove enclosed by these loops; and (3) favourable electrostatic contacts of basic side chains with negatively charged membrane phosphate groups.     

含有修饰SpA的重组生物膜的表征

通过ＬＢ膜技术将硬脂酸酯修饰的ＳｐＡ嵌入到气液界面的磷脂膜中，并沉积于疏水石英表面形成重组双层生物膜。此重组膜可作为免疫球蛋白的能诱导膜。结果表明，修饰ＳｐＡ表面硬脂酸数目的多少是影响该蛋白构象变化的主要原因。当共嵌入到磷脂单分子膜中时，膜中蛋白质密度增加，且呈现良好的成膜特性，重组膜中ＳｐＡ的活性下降与理解旨酸的修饰及ＳｐＡ在膜中嵌入作用有关。本文还讨论了膜中结合ＩｇＧ的状态。     

Electrochemical studies of Meldola bluemodified bilayer lipid membranes

Conclusion The voltammetric characteristics of MB transport across BLM are concurrent with the first type of dyes in ref. [5]. There is no peak appearing in the range of potential window and the transferring can be regarded as an irreversible one. The transport is tightly related to the structure of the membrane. Oxidized cholesterol and lipid negatively charged inhibit the transport of MB. The pH of the aqueous solution also influences the transport. When the pH of the aqueous solution is close to the isoelectric point of the lipid, the voltammetric response is obvious. When the pH is lower than that of the isoelectric point, the transport will be inhibited. These results can be explained by considering the appearance of net positive charges at the surface of the membrane at lower pH. The above investigations can provide important information for further studying the catalysis of MB to some certain bio-active substances (such as NADH) at the surface of biomembranes.     

Helix deformation is coupled to vectorial proton transport in the photocycle of bacteriorhodopsin

A wide variety of mechanisms are used to generate a proton-motive potential across cell membranes, a function lying at the heart of bioenergetics. Bacteriorhodopsin, the simplest known proton pump, provides a paradigm for understanding this process. Here we report, at 2.1 A resolution, the structural changes in bacteriorhodopsin immediately preceding the primary proton transfer event in its photocycle. The early structural rearrangements propagate from the protein's core towards the extracellular surface, disrupting the network of hydrogen-bonded water molecules that stabilizes helix C in the ground state. Concomitantly, a bend of this helix enables the negatively charged primary proton acceptor, Asp 85, to approach closer to the positively charged primary proton donor, the Schiff base. The primary proton transfer event would then neutralize these two groups, cancelling their electrostatic attraction and facilitating a relaxation of helix C to a less strained geometry. Reprotonation of the Schiff base by Asp 85 would thereby be impeded, ensuring vectorial proton transport. Structural rearrangements also occur near the protein's surface, aiding proton release to the extracellular medium.     

氧化甾醇-磷脂脂质囊泡的制备及特性研究

 氧化甾醇3,19–二羟基-胆甾烷-24-酮（DHCO）和3β,5α,6β-胆甾烷三醇（Triol）替代胆甾醇与大豆磷脂形成脂质囊泡的性质研究。采用乙醇注入法制备脂质囊泡,通过测定脂溶性及水溶性荧光探针在脂质囊泡中的荧光强度变化,考察囊泡膜流动性及通透性;通过测定脂质囊泡中游离DHCO及结合DHCO的浓度考察DHCO与磷脂的结合率;考察DHCO/磷脂比例、超声条件对脂质囊泡粒径大小和分布的影响。结果表明,DHCO、Triol与磷脂形成的脂质囊泡与胆甾烷（CHOL）-磷脂脂质囊泡的膜流动性无明显差异,但膜通透性稍有增大。DHCO与磷脂的结合率为8258%。DHCO、Triol与大豆磷脂经简单工艺即可形成脂质囊泡。可通过调节DHCO/磷脂比例、超声条件获得具理想粒径和外观的脂质囊泡。氧化甾醇数量庞大,其作为脂质囊泡的新型“流动性缓冲剂”有巨大的发展潜力。     

分子膜驱提高原油采收率机理研究

分子膜驱是一种新型的纳米膜驱提高原油采收率技术。采用多种实验手段,通过大量实验研究表明,分子膜驱的驱油机理有别于传统的化学驱（聚合物驱、表面活性剂驱、碱驱及其复合驱等油方法）。它是以水溶液为传递介质,膜剂分子依靠静电相互作用为成膜动力,膜剂有效分子沉积在呈负电性的岩石表面,形成纳米级超薄膜,改变了储层表面性质和与原油的相互作用状态,使得注入流体在冲刷孔隙过程中,原油易于剥落和流动而被驱替液驱替出来,达到提高采收率的目的。分子膜剂作用机理主要表现在吸附作用、润湿性改变、扩散作用、毛细管自吸作用等几个方面。