Showing papers on "Membrane published in 2008"

Membrane lipids: where they are and how they behave.

Abstract: Throughout the biological world, a 30 A hydrophobic film typically delimits the environments that serve as the margin between life and death for individual cells. Biochemical and biophysical findings have provided a detailed model of the composition and structure of membranes, which includes levels of dynamic organization both across the lipid bilayer (lipid asymmetry) and in the lateral dimension (lipid domains) of membranes. How do cells apply anabolic and catabolic enzymes, translocases and transporters, plus the intrinsic physical phase behaviour of lipids and their interactions with membrane proteins, to create the unique compositions and multiple functionalities of their individual membranes?

Impermeable atomic membranes from graphene sheets.

Abstract: We demonstrate that a monolayer graphene membrane is impermeable to standard gases including helium. By applying a pressure difference across the membrane, we measure both the elastic constants and the mass of a single layer of graphene. This pressurized graphene membrane is the world's thinnest balloon and provides a unique separation barrier between 2 distinct regions that is only one atom thick.

Membrane recognition by phospholipid-binding domains.

Abstract: Many different globular domains bind to the surfaces of cellular membranes, or to specific phospholipid components in these membranes, and this binding is often tightly regulated. Examples include pleckstrin homology and C2 domains, which are among the largest domain families in the human proteome. Crystal structures, binding studies and analyses of subcellular localization have provided much insight into how members of this diverse group of domains bind to membranes, what features they recognize and how binding is controlled. A full appreciation of these processes is crucial for understanding how protein localization and membrane topography and trafficking are regulated in cells.

Parallel cylindrical water nanochannels in Nafion fuel-cell membranes.

Abstract: The structure of the Nafion ionomer used in proton-exchange membranes of H(2)/O(2) fuel cells has long been contentious. Using a recently introduced algorithm, we have quantitatively simulated previously published small-angle scattering data of hydrated Nafion. The characteristic 'ionomer peak' arises from long parallel but otherwise randomly packed water channels surrounded by partially hydrophilic side branches, forming inverted-micelle cylinders. At 20 vol% water, the water channels have diameters of between 1.8 and 3.5 nm, with an average of 2.4 nm. Nafion crystallites (approximately 10 vol%), which form physical crosslinks that are crucial for the mechanical properties of Nafion films, are elongated and parallel to the water channels, with cross-sections of approximately (5 nm)(2). Simulations for various other models of Nafion, including Gierke's cluster and the polymer-bundle model, do not match the scattering data. The new model can explain important features of Nafion, including fast diffusion of water and protons through Nafion and its persistence at low temperatures.

Surface-structure-regulated cell-membrane penetration by monolayer-protected nanoparticles

Abstract: Nanoscale objects are typically internalized by cells into membrane-bounded endosomes and fail to access the cytosolic cell machinery. Whereas some biomacromolecules may penetrate or fuse with cell membranes without overt membrane disruption, no synthetic material of comparable size has shown this property yet. Cationic nano-objects pass through cell membranes by generating transient holes, a process associated with cytotoxicity. Studies aimed at generating cell-penetrating nanomaterials have focused on the effect of size, shape and composition. Here, we compare membrane penetration by two nanoparticle 'isomers' with similar composition (same hydrophobic content), one coated with subnanometre striations of alternating anionic and hydrophobic groups, and the other coated with the same moieties but in a random distribution. We show that the former particles penetrate the plasma membrane without bilayer disruption, whereas the latter are mostly trapped in endosomes. Our results offer a paradigm for analysing the fundamental problem of cell-membrane-penetrating bio- and macro-molecules.

Direct Imaging of Lattice Atoms and Topological Defects in Graphene Membranes

Abstract: We present a transmission electron microscopy investigation of graphene membranes, crystalline foils with a thickness of only 1 atom. By using aberration-correction in combination with a monochromator, 1-A resolution is achieved at an acceleration voltage of only 80 kV. The low voltage is crucial for the stability of these membranes. As a result, every individual carbon atom in the field of view is detected and resolved. We observe a highly crystalline lattice along with occasional point defects. The formation and annealing of Stone-Wales defects is observed in situ. Multiple five- and seven-membered rings appear exclusively in combinations that avoid dislocations and disclinations, in contrast to previous observations on highly curved (tube- or fullerene-like) graphene surfaces.

Superwetting nanowire membranes for selective absorption

Abstract: The construction of nanoporous membranes is of great technological importance for various applications, including catalyst supports, filters for biomolecule purification, environmental remediation and seawater desalination. A major challenge is the scalable fabrication of membranes with the desirable combination of good thermal stability, high selectivity and excellent recyclability. Here we present a self-assembly method for constructing thermally stable, free-standing nanowire membranes that exhibit controlled wetting behaviour ranging from superhydrophilic to superhydrophobic. These membranes can selectively absorb oils up to 20 times the material's weight in preference to water, through a combination of superhydrophobicity and capillary action. Moreover, the nanowires that form the membrane structure can be re-suspended in solutions and subsequently re-form the original paper-like morphology over many cycles. Our results suggest an innovative material that should find practical applications in the removal of organics, particularly in the field of oil spill cleanup.

Membrane phosphatidylserine regulates surface charge and protein localization

Abstract: Electrostatic interactions with negatively charged membranes contribute to the subcellular targeting of proteins with polybasic clusters or cationic domains. Although the anionic phospholipid phosphatidylserine is comparatively abundant, its contribution to the surface charge of individual cellular membranes is unknown, partly because of the lack of reagents to analyze its distribution in intact cells. We developed a biosensor to study the subcellular distribution of phosphatidylserine and found that it binds the cytosolic leaflets of the plasma membrane, as well as endosomes and lysosomes. The negative charge associated with the presence of phosphatidylserine directed proteins with moderately positive charge to the endocytic pathway. More strongly cationic proteins, normally associated with the plasma membrane, relocalized to endocytic compartments when the plasma membrane surface charge decreased on calcium influx.

Mechanics of membrane fusion.

Abstract: Diverse membrane fusion reactions in biology involve close contact between two lipid bilayers, followed by the local distortion of the individual bilayers and reformation into a single, merged membrane. We consider the structures and energies of the fusion intermediates identified in experimental and theoretical work on protein-free lipid bilayers. On the basis of this analysis, we then discuss the conserved fusion-through-hemifusion pathway of merger between biological membranes and propose that the entire progression, from the close juxtaposition of membrane bilayers to the expansion of a fusion pore, is controlled by protein-generated membrane stresses.

Designing Carbon Nanotube Membranes for Efficient Water Desalination

Abstract: The transport of water and ions through membranes formed from carbon nanotubes ranging in diameter from 6 to 11 A is studied using molecular dynamics simulations under hydrostatic pressure and equilibrium conditions. Membranes incorporating carbon nanotubes are found to be promising candidates for water desalination using reverse osmosis, and the size and uniformity of tubes that is required to achieve a desired salt rejection is determined. By calculating the potential of mean force for ion and water translocation, we show that ions face a large energy barrier and will not pass through the narrower tubes studied ((5,5) and (6,6) “armchair” type tubes) but can pass through the wider (7,7) and (8,8) nanotubes. Water, however, faces no such impediment due to the formation of stable hydrogen bonds and crosses all of the tubes studied at very large rates. By measuring this conduction rate under a hydrostatic pressure difference, we show that membranes incorporating carbon nanotubes can, in principle, achieve ...

Mechanisms of membrane fusion: disparate players and common principles

Abstract: Membrane fusion can occur between cells, between different intracellular compartments, between intracellular compartments and the plasma membrane and between lipid-bound structures such as viral particles and cellular membranes In order for membranes to fuse they must first be brought together The more highly curved a membrane is, the more fusogenic it becomes We discuss how proteins, including SNAREs, synaptotagmins and viral fusion proteins, might mediate close membrane apposition and induction of membrane curvature to drive diverse fusion processes We also highlight common principles that can be derived from the analysis of the role of these proteins

Ion exclusion by sub-2-nm carbon nanotube pores

Abstract: Biological pores regulate the cellular traffic of a large variety of solutes, often with high selectivity and fast flow rates. These pores share several common structural features: the inner surface of the pore is frequently lined with hydrophobic residues, and the selectivity filter regions often contain charged functional groups. Hydrophobic, narrow-diameter carbon nanotubes can provide a simplified model of membrane channels by reproducing these critical features in a simpler and more robust platform. Previous studies demonstrated that carbon nanotube pores can support a water flux comparable to natural aquaporin channels. Here, we investigate ion transport through these pores using a sub-2-nm, aligned carbon nanotube membrane nanofluidic platform. To mimic the charged groups at the selectivity region, we introduce negatively charged groups at the opening of the carbon nanotubes by plasma treatment. Pressure-driven filtration experiments, coupled with capillary electrophoresis analysis of the permeate and feed, are used to quantify ion exclusion in these membranes as a function of solution ionic strength, pH, and ion valence. We show that carbon nanotube membranes exhibit significant ion exclusion that can be as high as 98% under certain conditions. Our results strongly support a Donnan-type rejection mechanism, dominated by electrostatic interactions between fixed membrane charges and mobile ions, whereas steric and hydrodynamic effects appear to be less important.

Dynamic multiple-target tracing to probe spatiotemporal cartography of cell membranes.

Abstract: Although the highly dynamic and mosaic organization of the plasma membrane is well-recognized, depicting a resolved, global view of this organization remains challenging. We present an analytical single-particle tracking (SPT) method and tool, multiple-target tracing (MTT), that takes advantage of the high spatial resolution provided by single-fluorophore sensitivity. MTT can be used to generate dynamic maps at high densities of tracked particles, thereby providing global representation of molecular dynamics in cell membranes. Deflation by subtracting detected peaks allows detection of lower-intensity peaks. We exhaustively detected particles using MTT, with performance reaching theoretical limits, and then reconnected trajectories integrating the statistical information from past trajectories. We demonstrate the potential of this method by applying it to the epidermal growth factor receptor (EGFR) labeled with quantum dots (Qdots), in the plasma membrane of live cells. We anticipate the use of MTT to explore molecular dynamics and interactions at the cell membrane.

Crowding Effects on Diffusion in Solutions and Cells

Abstract: We review the effects of molecular crowding on solute diffusion in solution and in cellular aqueous compartments and membranes. Anomalous diffusion, in which mean squared displacement does not increase linearly with time, is predicted in simulations of solute diffusion in media crowded with fixed or mobile obstacles, or when solute diffusion is restricted or accelerated by a variety of geometric or active transport processes. Experimental measurements of solute diffusion in solutions and cellular aqueous compartments, however, generally show Brownian diffusion. In cell membranes, there are examples of both Brownian and anomalous diffusion, with the latter likely produced by lipid-protein and protein-protein interactions. We conclude that the notion of universally anomalous diffusion in cells as a consequence of molecular crowding is not correct and that slowing of diffusion in cells is less marked than has been generally assumed.

Template-directed Synthesis of a Genetic Polymer in a Model Protocell

Abstract: The phospholipids that form the membranes of modern cells present a formidable barrier to polar and charged molecules, necessitating complex channels and pumps to permit the exchange of molecules with the external environment. This presents a problem when trying to imagine what a primitive cell would have looked like earlier in the evolution of life. With no transport machinery, how could simple cells take in complex nutrients? A possible answer comes in the form of a model 'protocell' produced by a team at Harvard. Fatty acids and their derivatives are attractive candidates as components of early protocell membranes as they are simple amphiphiles that form bilayer membrane vesicles. A proto-cell equipped with such a membrane is found to be able to take in charged molecules such as nucleotides, while retaining longer genetic polymers made from them inside. The membranes of modern cells are made of phospholipids, which are formidable barriers to the uptake of polar and charged molecules, a challenge to our understanding of the origins of cellular life. Membranes made of simple amphiphiles, such as fatty acids, allow the passage of charged molecules (such as nucleotides), while retaining longer genetic polymers made from them inside such protocells. Primitive cells could thus have acquired complex nutrients from their environment in the absence of any transport machinery. Contemporary phospholipid-based cell membranes are formidable barriers to the uptake of polar and charged molecules ranging from metal ions to complex nutrients. Modern cells therefore require sophisticated protein channels and pumps to mediate the exchange of molecules with their environment. The strong barrier function of membranes has made it difficult to understand the origin of cellular life and has been thought to preclude a heterotrophic lifestyle for primitive cells. Although nucleotides can cross dimyristoyl phosphatidylcholine membranes through defects formed at the gel-to-liquid transition temperature1,2, phospholipid membranes lack the dynamic properties required for membrane growth. Fatty acids and their corresponding alcohols and glycerol monoesters are attractive candidates for the components of protocell membranes because they are simple amphiphiles that form bilayer membrane vesicles3,4,5 that retain encapsulated oligonucleotides3,6 and are capable of growth and division7,8,9. Here we show that such membranes allow the passage of charged molecules such as nucleotides, so that activated nucleotides added to the outside of a model protocell spontaneously cross the membrane and take part in efficient template copying in the protocell interior. The permeability properties of prebiotically plausible membranes suggest that primitive protocells could have acquired complex nutrients from their environment in the absence of any macromolecular transport machinery; that is, they could have been obligate heterotrophs.

Self-Assembly of Large and Small Molecules into Hierarchically Ordered Sacs and Membranes

Abstract: We report here the self-assembly of macroscopic sacs and membranes at the interface between two aqueous solutions, one containing a megadalton polymer and the other, small self-assembling molecules bearing opposite charge. The resulting structures have a highly ordered architecture in which nanofiber bundles align and reorient by nearly 90° as the membrane grows. The formation of a diffusion barrier upon contact between the two liquids prevents their chaotic mixing. We hypothesize that growth of the membrane is then driven by a dynamic synergy between osmotic pressure of ions and static self-assembly. These robust, self-sealing macroscopic structures offer opportunities in many areas, including the formation of privileged environments for cells, immune barriers, new biological assays, and self-assembly of ordered thick membranes for diverse applications.

Wide varieties of cationic nanoparticles induce defects in supported lipid bilayers.

Abstract: Nanoparticles with widely varying physical properties and origins (spherical versus irregular, synthetic versus biological, organic versus inorganic, flexible versus rigid, small versus large) have been previously noted to translocate across the cell plasma membrane. We have employed atomic force microscopy to determine if the physical disruption of lipid membranes, formation of holes and/or thinned regions, is a common mechanism of interaction between these nanoparticles and lipids. It was found that a wide variety of nanoparticles, including a cell penetrating peptide (MSI-78), a protein (TAT), polycationic polymers (PAMAM dendrimers, pentanol-core PAMAM dendrons, polyethyleneimine, and diethylaminoethyl-dextran), and two inorganic particles (Au-NH2, SiO2-NH2), can induce disruption, including the formation of holes, membrane thinning, and/or membrane erosion, in supported lipid bilayers.

Spatial regulators for bacterial cell division self-organize into surface waves in vitro.

Abstract: In the bacterium Escherichia coli, the Min proteins oscillate between the cell poles to select the cell center as division site. This dynamic pattern has been proposed to arise by self-organization of these proteins, and several models have suggested a reaction-diffusion type mechanism. Here, we found that the Min proteins spontaneously formed planar surface waves on a flat membrane in vitro. The formation and maintenance of these patterns, which extended for hundreds of micrometers, required adenosine 5'-triphosphate (ATP), and they persisted for hours. We present a reaction-diffusion model of the MinD and MinE dynamics that accounts for our experimental observations and also captures the in vivo oscillations.

The molecular face of lipid rafts in model membranes.

Abstract: Cell membranes contain a large number of different lipid species. Such a multicomponent mixture exhibits a complex phase behavior with regions of structural and compositional heterogeneity. Especially domains formed in ternary mixtures, composed of saturated and unsaturated lipids together with cholesterol, have received a lot of attention as they may resemble raft formation in real cells. Here we apply a simulation model to assess the molecular nature of these domains at the nanoscale, information that has thus far eluded experimental determination. We are able to show the spontaneous separation of a saturated phosphatidylcholine (PC)/unsaturated PC/cholesterol mixture into a liquid-ordered and a liquid-disordered phase with structural and dynamic properties closely matching experimental data. The near-atomic resolution of the simulations reveals remarkable features of both domains and the boundary domain interface. Furthermore, we predict the existence of a small surface tension between the monolayer leaflets that drives registration of the domains. At the level of molecular detail, raft-like lipid mixtures show a surprising face with possible implications for many cell membrane processes.

Structure and function of bordered pits: new discoveries and impacts on whole-plant hydraulic function

Abstract: Bordered pits are cavities in the lignified cell walls of xylem conduits (vessels and tracheids) that are essential components in the water-transport system of higher plants. The pit membrane, which lies in the center of each pit, allows water to pass between xylem conduits but limits the spread of embolism and vascular pathogens in the xylem. Averaged across a wide range of species, pits account for > 50% of total xylem hydraulic resistance, indicating that they are an important factor in the overall hydraulic efficiency of plants. The structure of pits varies dramatically across species, with large differences evident in the porosity and thickness of pit membranes. Because greater porosity reduces hydraulic resistance but increases vulnerability to embolism, differences in pit structure are expected to correlate with trade-offs between efficiency and safety of water transport. However, trade-offs in hydraulic function are influenced both by pit-level differences in structure (e.g. average porosity of pit membranes) and by tissue-level changes in conduit allometry (average length, diameter) and the total surface area of pit membranes that connects vessels. In this review we address the impact of variation in pit structure on water transport in plants from the level of individual pits to the whole plant.