Role of a highly conserved bacterial protein in outer membrane protein assembly

After transport across the cytoplasmic membrane, bacterial outer membrane proteins are assembled into the outer membrane. Meningococcal Omp85 is a highly conserved protein in Gram-negative bacteria, and its homolog Toc75 is a component of the chloroplast protein-import machinery. Omp85 appeared to be essential for viability, and unassembled forms of various outer membrane proteins accumulated upon Omp85 depletion. Immunofluorescence microscopy revealed decreased surface exposure of outer membrane proteins, which was particularly apparent at the cell-division planes. Thus, Omp85 is likely to play a role in outer membrane protein assembly.     

Electrical output of a receptor membrane

The electrical output of the receptor membrane of the nonmyelinated ending of Pacinian corpuscles is a function of the electrical gradients across the receptor membrane. The generator potential of the receptor membrane in response to equal mechanical stimuli varies linearly with the intensity of polarizing currents passed through the membrane. The production of a generator potential leaves a refractory state in the receptor membrane which is independent of the amount of charge transferred across the membrane but is dependent on a factor related to the strength of the stimulus which produced the response.     

Single cell layered heart: electromechanical properties of the heart of Boltenia ovifera

The heart of Boltenia ovifera (the sea potato) is a tubular structure formed by a single layer of myocardial cells. Electron microscopic studies show that each cell contains a single myofibril located adjacent to the luminal surface of the cell. Electrical and mechanical measurement of a cannulated perfused heart demonstrate that only the luminal membrane is excitabble and elicits contraction on depolarization. Calcium and magnesium exert antagonistic effects on tension, and potassium depolarizes the myocardium and produces contractures when the luminal membrane is exposed to various concentrations of these ions. The extraluminl membrane does not respond electrically or mechanically to calcium magnesium or potassium, and its potential seems to be effectively "clamped" by the luminal membrane. Functionaly, therefore, this heart consists of a single active membrane with the adjacent contractile apparatus.     

Signal for attachment of a phospholipid membrane anchor in decay accelerating factor

Decay accelerating factor (DAF) belongs to a novel group of membrane proteins anchored to the cell surface by a glycophospholipid membrane anchor that is covalently attached to the carboxyl terminus of the protein. The last 37 amino acids of membrane DAF, when fused to the carboxyl terminus of a secreted protein, are sufficient to target the fusion protein to the plasma membrane by means of a glycophospholipid anchor. This approach provides a novel means of targeting proteins to the cell-surface membrane.     

Restricted lateral diffusion of PH-20, a PI-anchored sperm membrane protein

The rate of lateral diffusion of integral membrane proteins is constrained in cells, but the constraining factors for most membrane proteins have not been defined. PH-20, a sperm surface protein involved in sperm-egg adhesion, was shown to be anchored in the plasma membrane by attachment to the lipid phosphatidylinositol and to have a diffusion rate that is highly restricted on testicular sperm, being more than a thousand times slower than lipid diffusion. These results support the hypothesis that lateral mobility of a membrane protein can be regulated exclusively by interactions of its ectodomain.     

Outer membrane active transport: structure of the BtuB:TonB complex

In Gram-negative bacteria, the import of essential micronutrients across the outer membrane requires a transporter, an electrochemical gradient of protons across the inner membrane, and an inner membrane protein complex (ExbB, ExbD, TonB) that couples the proton-motive force to the outer membrane transporter. The inner membrane protein TonB binds directly to a conserved region, called the Ton-box, of the transporter. We solved the structure of the cobalamin transporter BtuB in complex with the C-terminal domain of TonB. In contrast to its conformations in the absence of TonB, the Ton-box forms a beta strand that is recruited to the existing beta sheet of TonB, which is consistent with a mechanical pulling model of transport.     

Structure of membranes: reaction of red blood cell membranes with phospholipase C

Treatment of human red blood cell membranes with phospholipase C releases 68 to 74 percent of the total membrane phosphorus into solution, through hydrolysis of membrane phospholipids to diglycerides and water-soluble phosphorylated amines. In spite of this drastic change, the membrane remains intact in phase microscopy, and the average protein conformation in the membranes, as determined by circular dichroism measurements in the ultraviolet, is unaffected. These results are readily explained by a model of membrane structure that is stabilized by hydrophobic interactions and in which the polar and ionic heads of lipids are on the outer surfaces of the membrane, in contact with the bulk aqueous phase and accessible to the action of phospholipase C.     

壳聚糖作为膜材料的研究进展

壳聚糖既能进行化学改性,以其为材料所制得的膜又具有生物相容性和生物可降解性,是一种良好的膜材料。综述了以壳聚糖为主要原料制备膜及其膜材料的研究进展。     

运行方式对减缓SMBR膜污染的影响研究

膜污染是制约膜生物反应器大规模应用的瓶颈问题 ,研究考察了运行方式对减缓膜污染的影响。试验结果表明 ,反冲洗组件 B相对于对照组件 A膜通量平均增加 2 2 .30 % ,证实反冲洗操作能够使与膜结合力较小的污染物脱落 ,依靠反冲洗可以帮助膜组件拥有较长的寿命。建议在工程运行中根据工况 ,采用自动反冲洗装置。同时详细阐述了曝气冲刷对抗膜污染的作用机理 ,给出自行设计的膜组件结构单元的立体图与平面图     

硼对作物细胞膜功能影响的研究进展

综述了硼对植物细胞膜功能的影响机理，指出硼同根细胞膜对硼的吸收密切相关。敏感基因型作物对硼的大量积累和耐硼毒基因型作物对硼的少量积累，都与细胞膜的透性不同有关，并且还与细胞膜和细胞壁的组成相关。硼酸进入根皮细胞可能有两种不同的方式，通过细胞质膜磷脂双分子层的扩散和通过通道蛋白。硼高效品种和低效品种在硼的吸收、运输和再利用方面的差异是细胞膜的组成、细胞膜对硼的透性、膜通道蛋白及其转运效率差异的反映。     

Selective solubilization of a protein component of the red cell membrane

Approximately 20 percent of the membrane-bound protein of erythrocyte ghosts can be solubilized and obtained free of other membrane components by dialysis against adenosine triphosphate and 2-mercaptoethanol. This protein forms one major band on polyacrylamide gels and a single boundary in free-boundary electrophoresis, and it undergoes polymerization in the presence of divalent cations to form coiled filaments visible by electron microscopy. Antibodies to this membrane protein react specifically with red blood cells or their membrane ghosts but do not react with serum, erythrocyte cytoplasm, or other blood cells. The functional role of this protein is unknown, but it appears to be involved in maintaiining the structure of the red cell membrane. We suggest that this protein be called Spectrin since it is obtained from membrane ghosts.     

A glycophospholipid tail at the carboxyl terminus of the Thy-1 glycoprotein of neurons and thymocytes

Cell surface molecules of eukaryotic cells have been considered to be integrated into the membrane bilayer by a transmembrane protein sequence. The Thy-1 antigen of rodent thymocytes and brain was the first eukaryotic membrane molecule for which biochemical data clearly suggested membrane integration via a nonprotein tail. Direct evidence is now presented showing that a glycophospholipid structure is attached to the carboxyl-terminal cysteine residue and that 31 carboxyl-terminal amino acids predicted from the Thy-1 complementary DNA sequence are not present in the mature glycoprotein. These experimental results raise questions concerning signaling across a cell membrane since antibodies to Thy-1 can stimulate T lymphocytes to release lymphokines and undergo cell division.     

Observation of a membrane fusion intermediate structure

We report the observation of a phase of phospholipid that contains a structure similar to the commonly postulated interbilayer state that is crucial to membrane fusion. The widely accepted model for membrane fusion suggests that there is an intermediate state in which the two contacting monolayers become continuous via an hourglass-shaped structure called a stalk. Many efforts have been made to estimate the free energy for such a state in order to understand the functionality of membrane fusion proteins and to define key parameters in energy estimates. The observation of the stalk structure supports the stalk hypothesis for membrane fusion and enables the measurement of these parameters experimentally.     

Artificial membranes: a new type of cell for measuring diffusional resistance

A novel technique allows determination of membrane diffusivities and eliminates the complications of the fluid resistance to mass transfer on either side of the membrane. Results on the permeability of a dialysis membrane of the type used in artificial kidneys agree with previous data and are obtained in much shorter time. The measured activation energy for diffusion demonstrates that the transport of salt through the membrane occurs by the same mechanism as the diffusion of salt in water, but that only 10 percent of the membrane surface is effective.     

Calorimetric detection of a membrane-lipid phase transition in living cells

The membrane lipids in living Mycoplasma laidlawii exhibit a phase transition characteristic of that from crystal to liquid crystal within the bilayer conformation. The transition occurs at the same temperature in viable organisms, membranes isolated from the organisms, and isolated membrane lipids. The enthalpy of the transition in the membrane is compared with that of an aqueous suspension of isolated membrane lipids. The result is consistent with presence of an extended lipid bilayer in the native membrane.     

The fluid mosaic model of the structure of cell membranes

A fluid mosaic model is presented for the gross organization and structure of the proteins and lipids of biological membranes. The model is consistent with the restrictions imposed by thermodynamics. In this model, the proteins that are integral to the membrane are a heterogeneous set of globular molecules, each arranged in an amphipathic structure, that is, with the ionic and highly polar groups protruding from the membrane into the aqueous phase, and the nonpolar groups largely buried in the hydrophobic interior of the membrane. These globular molecules are partially embedded in a matrix of phospholipid. The bulk of the phospholipid is organized as a discontinuous, fluid bilayer, although a small fraction of the lipid may interact specifically with the membrane proteins. The fluid mosaic structure is therefore formally analogous to a two-dimensional oriented solution of integral proteins (or lipoproteins) in the viscous phospholipid bilayer solvent. Recent experiments with a wide variety of techniqes and several different membrane systems are described, all of which abet consistent with, and add much detail to, the fluid mosaic model. It therefore seems appropriate to suggest possible mechanisms for various membrane functions and membrane-mediated phenomena in the light of the model. As examples, experimentally testable mechanisms are suggested for cell surface changes in malignant transformation, and for cooperative effects exhibited in the interactions of membranes with some specific ligands. Note added in proof: Since this article was written, we have obtained electron microscopic evidence (69) that the concanavalin A binding sites on the membranes of SV40 virus-transformed mouse fibroblasts (3T3 cells) are more clustered than the sites on the membranes of normal cells, as predicted by the hypothesis represented in Fig. 7B. T-here has also appeared a study by Taylor et al. (70) showing the remarkable effects produced on lymphocytes by the addition of antibodies directed to their surface immunoglobulin molecules. The antibodies induce a redistribution and pinocytosis of these surface immunoglobulins, so that within about 30 minutes at 37 degrees C the surface immunoglobulins are completely swept out of the membrane. These effects do not occur, however, if the bivalent antibodies are replaced by their univalent Fab fragments or if the antibody experiments are carried out at 0 degrees C instead of 37 degrees C. These and related results strongly indicate that the bivalent antibodies produce an aggregation of the surface immunoglobulin molecules in the plane of the membrane, which can occur only if the immunoglobulin molecules are free to diffuse in the membrane. This aggregation then appears to trigger off the pinocytosis of the membrane components by some unknown mechanism. Such membrane transformations may be of crucial importance in the induction of an antibody response to an antigen, as well as iv other processes of cell differentiation.     

Adrenaline: mechanism of action on the pacemaker potential in cardiac Purkinje fibers

The pacemaker potential in Purkinje fibers is generated by a slow fall in potassium current which allows the inward background currents to depolarize the membrane. Adrenaline shifts the relation between activation of the potassium current and membrane potential in a depolarizing direction. Consequently, during the pacemaker potential, the potassium current falls more rapidly to lower values and the inward currents then depolarize the membrane more quickly. The shift in the potassium activation curve produced by adrenaline is large compared to that produced by calcium ions. The molecular action of adrenaline may involve either a large change in the surface charge of the membrane or a change in the dependence of the potassium permeability on the local electric field.     

Structure of C8alpha-MACPF reveals mechanism of membrane attack in complement immune defense

Membrane attack is important for mammalian immune defense against invading microorganisms and infected host cells. Proteins of the complement membrane attack complex (MAC) and the protein perforin share a common MACPF domain that is responsible for membrane insertion and pore formation. We determined the crystal structure of the MACPF domain of complement component C8alpha at 2.5 angstrom resolution and show that it is structurally homologous to the bacterial, pore-forming, cholesterol-dependent cytolysins. The structure displays two regions that (in the bacterial cytolysins) refold into transmembrane beta hairpins, forming the lining of a barrel pore. Local hydrophobicity explains why C8alpha is the first complement protein to insert into the membrane. The size of the MACPF domain is consistent with known C9 pore sizes. These data imply that these mammalian and bacterial cytolytic proteins share a common mechanism of membrane insertion.     

农田残膜机械回收膜土分离装置设计与试验

在我国南方农田残膜回收作业过程中,因土壤粘性强导致残膜夹带土壤多,残膜极易缠绕在机具上,影响机具正常工作。针对该问题设计了一种膜土分离装置,该装置主要由挑膜弹齿、滚筒和凸轮压板机构组成,可在挑膜过程中去除膜上表层土壤并在送膜过程中去除膜下土壤,通过试验分析得到影响膜土分离的主要因素为机具前进速度、挑膜转速和滚筒安装倾角。确定以上述三个因素为试验因素,以含土率和缠膜率为响应值,进行三因素三水平响应面试验。利用Desige-Expert软件建立各因素与缠膜率、含土率的回归模型,分析各因素对回收效率的影响,结果表明,各因素对含土率的显著性顺序为：机具前进速度＞挑膜转速＞滚筒安装倾角;对缠膜率的显著性顺序为：滚筒安装倾角＞挑膜转速＞机具前进速度;同时,对试验因素进行优化,得到因素的最佳组合为：机具前进速度为1.12 m·s^-1 ,挑膜转速为92.00 r·min-1,滚筒安装倾角为17.00°,此时的含土率为13.00%,缠膜率为1.70%。通过土槽模拟试验,设置上述最佳工作参数组合,得到含土率为13.45%,缠膜率为1.78%,验证结果与模型优化结果相对误差都小于5%。该膜土分离装置可以解决南方粘性土壤膜土分离困难、残膜率高的问题,为后续膜土分离装置及残膜回收机的设计与优化提供参考。     

Partitioning of lipid-modified monomeric GFPs into membrane microdomains of live cells

Many proteins associated with the plasma membrane are known to partition into submicroscopic sphingolipid- and cholesterol-rich domains called lipid rafts, but the determinants dictating this segregation of proteins in the membrane are poorly understood. We suppressed the tendency of Aequorea fluorescent proteins to dimerize and targeted these variants to the plasma membrane using several different types of lipid anchors. Fluorescence resonance energy transfer measurements in living cells revealed that acyl but not prenyl modifications promote clustering in lipid rafts. Thus the nature of the lipid anchor on a protein is sufficient to determine submicroscopic localization within the plasma membrane.