Langmuir-blodgett films of a functionalized molecule with cross-sectional mismatch between head and tail

A functionalized surfactant has been investigated as floating monolayers by synchrotron x-ray diffraction and as bilayers transferred to solid supports by the Langmuir-Blodgett technique through atomic force microscopy. The transfer process is accompanied by an increase of the unit cell area (about 17 percent) and by an increase of the average domain diameter of nanometer-scale domains (about three times). The unit cell area of the floating monolayer corresponds to close packing of the head groups and a noncharacteristic packing of the tifted alkyl chains. The larger unit cell area of the bilayer film is consistent with a particular ordered packing of the alkyl chains, leaving free space for the head groups.     

Identification of synaptophysin as a hexameric channel protein of the synaptic vesicle membrane

The quaternary structure and functional properties of synaptophysin, a major integral membrane protein of small presynaptic vesicles, were investigated. Cross-linking and sedimentation studies indicate that synaptophysin is a hexameric homo-oligomer, which in electron micrographs exhibits structural features common to channel-forming proteins. On reconstitution into planar lipid bilayers, purified synaptophysin displays voltage-sensitive channel activity with an average conductance of about 150 picosiemens. Because specific channels and fusion pores have been implicated in vesicular uptake and release of secretory compounds, synaptophysin may have a role in these processes.     

The ninth component of complement and the pore-forming protein (perforin 1) from cytotoxic T cells: structural, immunological, and functional similarities

The ninth component of complement (C9) and the pore-forming protein (PFP or perforin) from cytotoxic T lymphocytes polymerize to tubular lesions having an internal diameter of 100 A and 160 A, respectively, when bound to lipid bilayers. Polymerized C9, assembled by slow spontaneous or rapid Zn2+-induced polymerization, and polyperforin, which is assembled only in the presence of Ca2+, constitute large aqueous pores that are stable, nonselective for solutes, and insensitive to changes of membrane potential. Monospecific polyclonal antibodies to purified C9 and PFP show cross-reactivity, suggesting structural homology between the two molecules. The structural and functional homologies between these two killer molecules imply an active role for pore formation during cell lysis.     

Calcium channels in planar lipid bilayers: insights into mechanisms of ion permeation and gating

Electrophysiological recordings were used to analyze single calcium channels in planar lipid bilayers after membranes from bovine cardiac sarcolemmal vesicles had been incorporated into the bilayer. In these cell-free conditions, channels in the bilayer showed unitary barium or calcium conductances, gating kinetics, and pharmacological responses that were similar to dihydropyridine-sensitive calcium channels in intact cells. The open channel current varied in a nonlinear manner with voltage under asymmetric (that is, physiological) ionic conditions. However, with identical solutions on both sides of the bilayer, the current-voltage relation was linear. In matched experiments, calcium channels from skeletal muscle T-tubules differed significantly from cardiac calcium channels in their conductance properties and gating kinetics.     

Spherical bilayer vesicles of fullerene-based surfactants in water: a laser light scattering study

The low solubility of fullerenes in aqueous solution limits their applications in biology. By appropriate substitution, the fullerenes can be transformed into stabilized anions that are water soluble and can form large aggregated structures. A laser light scattering study of the association behavior of the potassium salt of pentaphenyl fullerene (Ph5C60K) in water revealed that the hydrocarbon anions Ph5C60- associate into bilayers, forming stable spherical vesicles with an average hydrodynamic radius and a radius of gyration of about 17 nanometers at a very low critical aggregation concentration of less than 10(-7) moles per liter. The average aggregation number of associated particles in these large spherical vesicles is about 1.2 x 10(4).     

Spontaneous vesicle formation in aqueous mixtures of single-tailed surfactants

Spontaneous, single-walled, equilibrium vesicles can be prepared from aqueous mixtures of simple, commercially available, single-tailed cationic and anionic surfactants. Vesicle size, surface charge, or permeability can be readily adjusted by varying the ratio of anionic to cationic surfactant. Vesicle formation apparently results from the production of anion-cation surfactant pairs that then act as double-tailed zwitterionic surfactants. These vesicles are quite stable in comparison to conventional vesicles prepared by mechanical disruption of insoluble liquid crystalline dispersions.     

Lipid domains in fluid membranes: a quick-freeze differential scanning calorimetry study

The application of rapid-freezing techniques to differential scanning calorimetry (DSC) provides a new approach for understanding the organization of lipids in biomembranes. Use of quick-freeze DSC on membranes of mixed lipid composition supports the existence of nonrandom distributions of lipids (domains) in fluid bilayers. In addition to allowing investigations on the organization of lipids in fluid bilayers, the quick-freeze technique now allows calorimetric studies to be carried out on mammalian membranes which, because of their high cholesterol content, have not been previously amenable to study by DSC.     

Carbon isotope composition of carbonaceous matter from the precambrian of the witwatersrand system

Polymerized hydrocarbons occurring in the gold-uranium conglomerates of the Witwatersrand System (South Africa) show deltaC(13) values between -22.4 and -32.8 per mille, their isotopic composition thus falling into the range of sedimentary organic carbon. Accordingly, organic derivation of the material seems very probable. This conclusion is consistent with a model of the existence of organic evolution and biologic activity in times certainly older than 2.15 x 10(9) years.     

Living with clathrin: its role in intracellular membrane traffic

Clathrin polymerization at the cytoplasmic side of the plasma membrane forms coated pits and vesicles that mediate uptake of cell surface receptors. Clathrin-coated vesicles have also been implicated in protein export but definition of their precise role has been controversial. Recent advances in characterization of the clathrin subunits and additional coated vesicle components have identified molecular interactions involved in clathrin polymerization and coated vesicle formation, and have provided new approaches to investigating its function. These studies suggest that clathrin's role, in both inward and outward membrane traffic, is to facilitate receptor transport by a concentration and sorting process that initiates targeting to specific intracellular compartments.     

Helical structures of ESCRT-III are disassembled by VPS4

During intracellular membrane trafficking and remodeling, protein complexes known as the ESCRTs (endosomal sorting complexes required for transport) interact with membranes and are required for budding processes directed away from the cytosol, including the budding of intralumenal vesicles to form multivesicular bodies; for the budding of some enveloped viruses; and for daughter cell scission in cytokinesis. We found that the ESCRT-III proteins CHMP2A and CHMP3 (charged multivesicular body proteins 2A and 3) could assemble in vitro into helical tubular structures that expose their membrane interaction sites on the outside of the tubule, whereas the AAA-type adenosine triphosphatase VPS4 could bind on the inside of the tubule and disassemble the tubes upon adenosine triphosphate hydrolysis. CHMP2A and CHMP3 copolymerized in solution, and their membrane targeting was cooperatively enhanced on planar lipid bilayers. Such helical CHMP structures could thus assemble within the neck of an inwardly budding vesicle, catalyzing late steps in budding under the control of VPS4.     

Self-assembly of flat nanodiscs in salt-free catanionic surfactant solutions

Discs of finite size are a very rare form of stable surfactant self-assembly. It is shown that mixing of two oppositely charged single-chain surfactants can produce rigid nanodiscs as well as swollen lamellar liquid crystals with frozen bilayers. The crucial requirement for obtaining nanodisc self-assembly is the use of H+ and OH- as counterions. These counterions then form water and lower the conductivity to 10 microsiemens per centimeter. In the case of cationic component excess, a dilute solution of nanodiscs is in thermodynamic equilibrium with a lamellar phase. The diameter of the cationic nanodiscs is continuously adjustable from a few micrometers to 30 nanometers, with the positive charge located mainly around the edges.     

Surfactant-mediated two-dimensional crystallization of colloidal crystals

Colloidal particles can form unexpected two-dimensional ordered colloidal crystals when they interact with surfactants of the opposite charge. Coulomb interactions lead to self-limited adsorption of the particles on the surface of vesicles formed by the surfactants. The adsorbed particles form ordered but fluid rafts on the vesicle surfaces, and these ultimately form robust two-dimensional crystals. This use of attractive Coulomb interaction between colloidal particles and surfactant structures offers a potential new route to self-assembly of ordered colloidal structures.     

Preferred microtubules for vesicle transport in lobster axons

The hypothesis that transported vesicles are preferentially associated with a subclass of microtubules has been tested in lobster axons. A cold block was used to collect moving vesicles in these axons; this treatment caused the vesicles to accumulate in files along some of the microtubules. Quantitative analysis of the number of vesicles associated with microtubule segments indicated that lobster axons have two distinct populations of microtubules--transport microtubules that are the preferred substrates for vesicle transport and architectural microtubules that contribute to axonal structure.     

微生物源脂肽的研究进展

脂肽可以由多种微生物产生,其化学结构多样,具有降低表面和界面处的张力的能力。脂肽作为表面活性剂的功能已经众所周知,并得到了广泛的应用。但它们在抗菌、抗病毒、抗肿瘤、溶血栓和杀虫等方面的功能还有待深入研究,具有良好的应用潜力和推广前景。主要回顾和总结了脂肽的生产、生物合成、提纯和应用,为同领域相关研究提供参考。     

Experimental models of primitive cellular compartments: encapsulation, growth, and division

The clay montmorillonite is known to catalyze the polymerization of RNA from activated ribonucleotides. Here we report that montmorillonite accelerates the spontaneous conversion of fatty acid micelles into vesicles. Clay particles often become encapsulated in these vesicles, thus providing a pathway for the prebiotic encapsulation of catalytically active surfaces within membrane vesicles. In addition, RNA adsorbed to clay can be encapsulated within vesicles. Once formed, such vesicles can grow by incorporating fatty acid supplied as micelles and can divide without dilution of their contents by extrusion through small pores. These processes mediate vesicle replication through cycles of growth and division. The formation, growth, and division of the earliest cells may have occurred in response to similar interactions with mineral particles and inputs of material and energy.     

Osmotic swelling of phospholipid vesicles causes them to fuse with a planar phospholipid bilayer membrane

Fusion of phospholipid vesicles with planar bilayer membranes occurs if the vesicles that contact the planar membrane swell osmotically after the replacement in their medium of an impermeant solute by a permeant one. This finding directly demonstrates that osmotic swelling is a driving force for vesicle-planar membrane fusion. The method used to induce vesicle swelling and fusion may have relevance for biological systems.     

Calcium uptake by isolated sarcoplasmic reticulum treated with dithiothreitol

Isolated vesicles of the sarcoplasmic reticulum are known to take up calcium when provided with magnesium adenosine triphosphate as an energy source. Preparations of high activity are obtained by keeping the vesicles in 5 millimolar dithiothreitol (a reagent that reduces disulfide groups), and these preparations retain activity for a week or longer. The highly active preparations lend themselves to a spectrophotometric method for following calcium uptake, and continuous uptake kinetics are readily obtained. Calcium uptake appears to follow Michaelis-Menten kinetics (K(m) = 8x10(-6); V(mas), = 7x10(-7) mole per second per milligram of protein). These preparations are also useful for studying the effects of inhibitors of uptake, such as quinine. When extrapolated to the intact muscle, the results from these isolated vesicles should give a better estimate than has been available of the actual rates of calcium uptake and of the physiological effect of inhibitors of uptake.     

The structures of COPI-coated vesicles reveal alternate coatomer conformations and interactions

Transport between compartments of eukaryotic cells is mediated by coated vesicles. The archetypal protein coats COPI, COPII, and clathrin are conserved from yeast to human. Structural studies of COPII and clathrin coats assembled in vitro without membranes suggest that coat components assemble regular cages with the same set of interactions between components. Detailed three-dimensional structures of coated membrane vesicles have not been obtained. Here, we solved the structures of individual COPI-coated membrane vesicles by cryoelectron tomography and subtomogram averaging of in vitro reconstituted budding reactions. The coat protein complex, coatomer, was observed to adopt alternative conformations to change the number of other coatomers with which it interacts and to form vesicles with variable sizes and shapes. This represents a fundamentally different basis for vesicle coat assembly.     

A G protein directly regulates mammalian cardiac calcium channels

A possible direct effect of guanine nucleotide binding (G) proteins on calcium channels was examined in membrane patches excised from guinea pig cardiac myocytes and bovine cardiac sarcolemmal vesicles incorporated into planar lipid bilayers. The guanosine triphosphate analog, GTP gamma S, prolonged the survival of excised calcium channels independently of the presence of adenosine 3',5'-monophosphate (cAMP), adenosine triphosphate, cAMP-activated protein kinase, and the protein kinase C activator tetradecanoyl phorbol acetate. A specific G protein, activated Gs, or its alpha subunit, purified from the plasma membranes of human erythrocytes, prolonged the survival of excised channels and stimulated the activity of incorporated channels. Thus, in addition to regulating calcium channels indirectly through activation of cytoplasmic kinases, G proteins can regulate calcium channels directly. Since they also directly regulate a subset of potassium channels, G proteins are now known to directly gate two classes of membrane ion channels.     

Surface-induced x-ray reflection visualization of membrane orientation and fusion into multibilayers

The fusion of lipid membranes at the air-water interface has been detected with the use of x-ray reflection as a high-resolution, surface-sensitive technique. The rate of this fusion for dimyristoylphosphatidylcholine (DMPC) bilayers is the highest at 29 degrees C, which coincides with the chain-melting phase-transition temperature for the top membrane layers. After 6 hours of incubation a stack of at least ten surface-ordered membrane bilayers in equilibrium with the bulk vesicle suspension is formed. Such fusion is thus surface-catalyzed but not restricted to the first surface layer. The process involves partial membrane dehydration near the solution surface which decreases toward the bulk.