Effect of ion concentration,solution and membrane permittivity on electric energy storage and capacitance

Bio-membranes as capacitors store electric energy, but their permittivity is low whereas the permittivity of surrounding solution is high. To evaluate the effective capacitance of the membrane/solution system and determine the electric energy stored within the membrane and in the solution, we estimated their electric variables using Poisson-Nernst-Planck simulations. We calculated membrane and solution capacitances from stored electric energy. The effective capacitance was calculated by fitting a six-capacitance model to charges (fixed and ion) and associated potentials, because it cannot be considered as a result of membrane and solution capacitance in series. The electric energy stored within the membrane (typically much smaller than that in the solution), depends on the membrane permittivity, but also on the external electric field, surface charge density, water permittivity and ion concentration. The effect on capacitances is more specific. Solution capacitance rises with greater solution permittivity or ion concentration, but the membrane capacitance (much smaller than solution capacitance) is only influenced by its permittivity. Interestingly, the effective capacitance is independent of membrane or solution permittivity, but rises as the ion concentration increases and surface charge becomes positive. Experimental estimates of membrane capacitance are thus not necessarily a reliable index of its surface area.     

Disulphide trapping of an in vivo energy-dependent conformation of Escherichia coli TonB protein

In Escherichia coli, the TonB system transduces the protonmotive force (pmf) of the cytoplasmic membrane to support a variety of transport events across the outer membrane. Cytoplasmic membrane proteins ExbB and ExbD appear to harvest pmf and transduce it to TonB. Experimental evidence suggests that TonB shuttles to the outer membrane, apparently to deliver conformationally stored potential energy to outer membrane transporters. In the most recent model, discharged TonB is then recycled to the cytoplasmic membrane to be re-energized by the energy coupling proteins, ExbB/D. It has been suggested that the carboxy-terminal 75 amino acids of active TonB could be represented by the rigid, strand-exchanged, dimeric crystal structure of the corresponding fragment. In contrast, recent genetic studies of alanine substitutions have suggested instead that in vivo the carboxy-terminus of intact TonB is dynamic and flexible. The biochemical studies presented here confirm and extend those results by demonstrating that individual cys substitution at aromatic residues in one monomeric subunit can form spontaneous dimers in vivo with the identical residue in the other monomeric subunit. Two energized TonBs appear to form a single cluster of 8-10 aromatic amino acids, including those found at opposite ends of the crystal structure. The aromatic cluster requires both the amino-terminal energy coupling domain of TonB, and ExbB/D (and cross-talk analogues TolQ/R) for in vivo formation. The large aromatic cluster is detected in cytoplasmic membrane-, but not outer membrane-associated TonB. Consistent with those observations, the aromatic cluster can form in the first half of the energy transduction cycle, before release of conformationally stored potential energy to ligand-loaded outer membrane transporters. The model that emerges is one in which, after input of pmf mediated through ExbB/D and the TonB transmembrane domain, the TonB carboxy-terminus can form a meta-stable high-energy conformation that is not represented by the crystal structure of the carboxy-terminus.     

Pore-forming toxins induce multiple cellular responses promoting survival

Pore-forming toxins (PFTs) are secreted proteins that contribute to the virulence of a great variety of bacterial pathogens. They inflict one of the more disastrous damages a target cell can be exposed to: disruption of plasma membrane integrity. Since this is an ancient form of attack, which bears similarities to mechanical membrane damage, cells have evolved response pathways to these perturbations. Here, it is reported that PFTs trigger very diverse yet specific response pathways. Many are triggered by the decrease in cytoplasmic potassium, which thus emerges as a central regulator. Upon plasma membrane damage, cells activate signalling pathways aimed at restoring plasma membrane integrity and ion homeostasis. Interestingly these pathways do not require protein synthesis. Cells also trigger signalling cascades that allow them to enter a quiescent-like state, where minimal energy is consumed while waiting for plasma membrane damage to be repaired. More specifically, protein synthesis is arrested, cytosolic constituents are recycled by autophagy and energy is stored in lipid droplets.     

Red cell membrane crenation: a macromodel of the echinocyte I

A physical macromodel of the erythrocyte membrane was constructed to investigate the effect of intrinsic membrane precurvature on a unique non-isotropic, composite material that dissipated shear energy but stored bending energy. The intrinsic precurvature of the material could be modified from strong positive precurvature through neutral to strong negative precurvature. A spherical shell constructed of the negatively precurved variant of this non-isotropic material spontaneously assumed the characteristic shape of the echinocyte I.     

Tensile loading characteristics of hydrogen stored carbon nanotubes in PEM fuel cell operating conditions using molecular dynamics simulation

Mechanical characteristics of hydrogen stored single walled carbon nanotube (SWCNT) in proton exchange membrane fuel cell (PEMFC) operating conditions are analysed in this work using molecular dynamics simulation method. The investigation of mechanical characteristics of hydrogen stored SWCNT is critical in determining the lifetime and stability of SWCNT-based membranes used in PEMFC. The study provides a comprehensive analysis on the effects of geometry, vacancy defects and PEMFC operating temperature on the mechanical properties of hydrogen stored SWCNT. The findings show that the mechanical strength of the hydrogen stored SWCNT can be enhanced by deploying a bigger armchair SWCNT. Furthermore, increase in operating temperature of PEMFC reduces the mechanical resistance of hydrogen stored SWCNT, which however can be overcome by suitably introducing vacancy defects in the SWCNT geometry. This has provided potential way of increasing the hydrogen storage capacity of SWCNT which is very useful for onboard application of PEMFC. It is anticipated that the findings obtained from this paper will have a paramount importance in the field of hydrogen energy fuel cell technology and further compliment the potential applications of SWCNTs as promising candidates for applications in fuel cells and energy storage devices.     

Adenilate pool and thylakoid ATP-synthase content in pea leaves under clinorotation

It is known that plant resistance to stress factors is connected with energy metabolism. The energy stored in the process of photophosphorylation in the form of ATP is used then to support respiration, transpiration, organic compound synthesis, growth and development as well as to restore cell structure after its damage under extremal environmental factors. Transformation of light energy into chemical energy of ATP is catalyzed by thylakoid membrane enzymatic complex of ATPsynthase-CF1CF0. Its activity and amount in the thylakoid membrane depends on plant growth conditions. The aim of this work was investigation of clinorotation effect on light-induced dynamics of adenyl nucleotides (AMP, ADP and ATP) and estimation of CF1CF0 content in thylakoids of pea leaves grown under slow clinorotation and vertical control.     

Heat production non-myelinated nerves.

Experiments with the C fibres of the rabbit vagus nerve have established that heat is evolved during the depolarizing phase of the action potential and is absorbed during the repolarizing phase. Subsequent studies using the pike olfactory nerve indicate that the heat production begins at a high rate very early in the depolarizing phase and is completed in advance of the peak of the spike. This would be expected if the heat arised from the energy released by the discharge of the membrane capacitance which varies as the square of the membrane potential; but estimates of the stored energy fall short of the observed heat production by a factor of two or three times. The prominent cooling phase suggests that a substantial part of the heat may arise from an entropy change. Such an entropy change would be expected to result from the change in the electrical stress in the dielectric of the membrane capacitance, and may thus be manifestation of reversible changes in the molecular architecture of the insulating matrix of the membrane.     

Photoacoustic photocalorimetry and spectroscopy of Halobacterium halobium purple membranes.

Enthalpy changes associated with intermediates of the photocycle of bacteriorhodopsin (bR) in light-adapted Halobacterium halobium purple membranes, and decay times of these intermediates, are obtained from photoacoustic measurements on purple membrane fragments. Our results, mainly derived from modulation frequency spectra, show changes in the amount of energy stored in the intermediates and in their decay times as a function of pH and/or salt concentration. Especially affected are the slowest step (endothermic) and a spectroscopically unidentified intermediate (both at pH 7). This effect is interpreted in terms of cation binding to the protein, conformational changes of which are thought to be connected with the endothermic process. Wavelength spectra are used to obtain heat dissipation spectra, which allow identification of wavelength regions with varying photoactivity, and estimation of the amounts of enthalpy stored in the photointermediates. Because of bleaching and accumulation of intermediates, however, and because of the small fraction of light energy stored during photocycle, quantitative information cannot be obtained. From photoacoustic wavelength spectra of purple membrane fragments equilibrated at 63% relative humidity, rise and decay times of the bR570 and M412 intermediates are calculated.     

Aggregation and disaggregation of red blood cells

The aggregation of red blood cells may be analyzed as an interaction of an adhesive surface energy and the elastic stored energy which results from deformation of the cell. The adhesive surface energy is the work required to separate a unit adhered area and is the resultant of adhesive forces due to the bridging molecules that bind the cells together and the electrostatic repulsion due to surface charge. The elastic strain energy in the case of the red blood is associated with the membrane elasticity only since the interior of the cell is liquid. The membrane elasticity is due both to bending stiffness and shear. The area expansion is small and may be neglected. These assumptions allow realistic computation of red cell shapes in rouleaux. The disaggregation of rouleaux requires an external force which must overcome the adhesive energy and also supply additional elastic energy of deformation. Depending on the geometry, the initial effect of elastic energy may tend to aid disaggregation. In a shear flow, the stresses on a suspended rouleau alternately tend to compress and to disaggregate the cells if they are free to rotate. This introduces a time dependence so that viscous effects due to the viscosity of the cell membrane, the cell cytoplasm and the external fluid may play a role in determining whether disaggregation proceeds to completion or not.     

Energy stored and dissipated in skeletal muscle basement membranes during sinusoidal oscillations.

We subjected single skeletal muscle cells from frog semitendinosus to sinusoidal oscillations that simulated the strain experienced as the cells near the end of passive extension and begin active contraction in slow swimming. Other cells from which the basement membrane was removed by enzymatic and mechanical procedures were tested identically. Effectiveness of the basement membrane removal technique was evaluated by electron microscopy, by an electrophoretic and lectin-binding assay for depletion of cell surface glycoproteins, and by confirmation by means of electrophoretic and immunologic analyses that major intracellular, cytoskeletal proteins were not disrupted. Measurements of maximum stress, maximum strain, and phase lag between these maxima enabled the complex modulus (dynamic stiffness) and loss tangent (relative viscous losses to elastic energy storage) to be calculated for each mechanically tested preparation. We also calculated the amounts of energy stored and dissipated in each preparation. These calculations indicate that cells with intact basement membranes have complex moduli significantly greater than those of cells without basement membranes, and that cells with basement membrane store significantly more elastic energy than basement membrane depleted cells. However, when subjected to identical sinusoidal strains, energy dissipation in cells with intact basement membranes is over three times greater than dissipation in cells without basement membrane. The relative magnitudes of energy losses to energy storage, called the specific loss, is nearly three times greater for intact cells than for basement membrane depleted cells. Basement membranes may thereby serve as a brake for slowing passive extension of muscle before contraction begins.     

ATP synthesis by F-type ATP synthase is obligatorily dependent on the transmembrane voltage.

ATP synthase is the universal enzyme that manufactures cellular ATP using the energy stored in a transmembrane ion gradient. This energy gradient has two components: the concentration difference (DeltapH or DeltapNa(+)) and the electrical potential difference DeltaPsi, which are thermodynamically equivalent. However, they are not kinetically equivalent, as the mitochondrial and bacterial ATP synthases require a transmembrane potential, DeltaPsi, but the chloroplast enzyme has appeared to operate on DeltapH alone. Here we show that, contrary to the accepted wisdom, the 'acid bath' procedure used to study the chloroplast enzyme develops not only a DeltapH but also a membrane potential, and that this potential is essential for ATP synthesis. Thus, for the chloroplast and other ATP synthases, the membrane potential is the fundamental driving force for their normal operation. We discuss the biochemical reasons for this phenomenon and a model that is consistent with these new experimental facts.     

The role of electrostatics in proton-conducting membrane protein complexes

Electrostatic interactions play a key role in the coupling of electron and proton transfer in membrane protein complexes during the conversion of the energy stored in sunlight or reduced substrates into biochemical energy via a transmembrane electrochemical proton potential. Principles of charge stabilization within membrane proteins are reviewed and discussed for photosynthetic reaction centers, cytochrome c oxidases, and diheme-containing quinol:fumarate reductases. The impact of X-ray structure-based electrostatic calculations on the functional interpretation of these structural coordinates, on providing new explanations for experimental observations, and for the design of more focused additional experiments is illustrated by a number of key examples.     

Energy-storage capacity of the mitochondrial proton-motive force

Resting state respiration of rat-liver mitochondria in the presence of oligomycin was rapidly blocked with cyanide and the dissipation of the membrane potential was followed with a tetraphenylphosphonium-sensitive electrode. From the rate of this dissipation and the electric capacitance of the mitochondrial membrane the energy stored in form of the membrane potential was calculated as about 7 microJ/mg protein. In the absence of oligomycin, dissipation of the membrane potential was slower, as it was partly compensated by proton ejection by mitochondrial ATPase hydrolyzing endogenous ATP. This allowed to calculate the total energy storage capacity of the proton-motive force. It amounted to the equivalence of 3.3 nmol ATP/mg protein or about 130 microJ/mg protein. The stoichiometry of proton-pumping ATPase utilizing endogenous ATP was estimated as three protons per molecule ATP.     

Vesicle formation in the Golgi apparatus

In this paper we examine the mechanics of vesicle budding from the Golgi apparatus. We propose a model for this process based on the notion that molecular surfactants can release the elastic energy stored in the lipid bilayer. The same physical process may drive other vesiculation processes, including coated vesicle formation and budding of enveloped viruses from the plasma membrane.     

Evolution of uphill electron transfer

The evolution of photosynthetic energy storage is considered. The primary event in primordial inorganic or organic photoreceptors was charge separation at the expense of light quantum energy. The subsequent improvement of energy storage was attained by separately channeling electrons and “holes” to prevent back reactions. The anisotropic arrangement of photoreceptors in the primary membrane caused a coupling of photochemical charge separation to subsequent ion dislocation and was a prerequisite of primary photophosphorylation. The gradual improvement of the molecular organization of photoreceptor units resulted in antenna and reaction center development. The “hole” was primary located on a peculiar photoreceptor form and the electron passed by tunneling through the chain of intermediate carriers (chlorophylls and pheophytins); thus long-lived charge separation was achieved. The use of the electrons and the “holes” stored in reaction centers for the functioning of the photosynthetic electron transfer chain was realized by cyclic and non-cyclic pathways when the coupling of two photochemical events became the more perfect mechanism to use water molecule as an ultimate electron donor. The appearance of primitive cells inevitably required the coupling of the solar energy conversion mechanism to the reproduction mechanism which used stored solar energy.     

Red blood cell membrane water permeability increases with length of ex vivo storage

Water transport across the red blood cell (RBC) membrane is an essential cell function that needs to be preserved during ex vivo storage. Progressive biochemical depletion during storage can result in significant conformational and compositional changes to the membrane. Characterizing the changes to RBC water permeability can help in evaluating the quality of stored blood products and aid in the development of improved methods for the cryopreservation of red blood cells. This study aimed to characterize the water permeability (L_p), osmotically inactive fraction (b), and Arrhenius activation energy (E_a) at defined storage time-points throughout storage and to correlate the observed results with other in vitro RBC quality parameters. RBCs were collected from age- and sex-matched blood donors. A stopped flow spectrophotometer was used to determine L_p and b by monitoring changes in hemoglobin autofluorescence when RBCs were exposed to anisotonic solutions. Experimental values of L_p were characterized at three different temperatures (4, 20 and 37 °C) to determine the E_a. Results showed that L_p, b, and E_a of stored RBCs significantly increase by day 21 of storage. Degradation of the RBC membrane with length of storage was seen as an increase in hemolysis and supernatant potassium, and a decrease in deformability, mean corpuscular hemoglobin concentration and supernatant sodium. RBC osmotic characteristics were shown to change with storage and correlate with changes in RBC membrane quality metrics. Monitoring water parameters is a predictor of membrane damage and loss of membrane integrity in ex vivo stored RBCs.     

The proton flux through the bacterial flagellar motor

Bacterial flagella are driven by a rotary motor that utilizes the free energy stored in the electrochemical proton gradient across the cytoplasmic membrane to do mechanical work. The flux of protons coupled to motor rotation was measured in Streptococcus and found to be directly proportional to motor speed. This supports the hypothesis that the movement of protons through the motor is tightly coupled to the rotation of its flagellar filament. Under this assumption the efficiency of energy conversion is close to unity at the low speeds encountered in tethered cells but only a few percent at the high speeds encountered in swimming cells. This difference appears to be due to dissipation by processes internal to the motor. The efficiency at high speeds exhibits a steep temperature dependence and a sizable deuterium solvent isotope effect.     

The generation of metabolic energy by solute transport

Secondary metabolic-energy-generating systems generate a proton motive force (pmf) or a sodium ion motive force (smf) by a process that involves the action of secondary transporters. The (electro)chemical gradient of the solute(s) is converted into the electrochemical gradient of protons or sodium ions. The most straightforward systems are the excretion systems by which a metabolic end product is excreted out of the cell in symport with protons or sodium ions (energy recycling). Similarly, solutes that were accumulated and stored in the cell under conditions of abundant energy supply may be excreted again in symport with protons when conditions become worse (energy storage). In fermentative bacteria, a proton motive force is generated by fermentation of weak acids, such as malate and citrate. The two components of the pmf, the membrane potential and the pH gradient, are generated in separate steps. The weak acid is taken up by a secondary transporter either in exchange with a fermentation product (precursor/product exchange) or by a uniporter mechanism. In both cases, net negative charge is translocated into the cell, thereby generating a membrane potential. Decarboxylation reactions in the metabolic breakdown of the weak acid consume cytoplasmic protons, thereby generating a pH gradient across the membrane. In this review, several examples of these different types of secondary metabolic energy generation will be discussed.     

Decreased ascorbic acid levels and brown core development in pears (Pyrus communis L. cv. Conference)

Changes in ascorbic acid content are measured in the cortex tissue of Conference pears stored at various compositions of carbon dioxide and oxygen. Enhanced carbon dioxide levels cause ascorbic acid concentrations to decline. Soon after ascorbic acid has declined below a certain value, browning of the core tissue can be observed. Reducing carbon dioxide levels before this value is reached causes ascorbic acid levels to increase again and prevents browning to a great extent. In preliminary experiments with a photoacoustic laser-based detection system, it was shown that pears that show browning produce ethane, which is most likely a result of membrane peroxidation. Storage conditions, ascorbic acid levels and browning in pears are discussed in relation to diffusion characteristics, energy metabolism and energy maintenance levels of the fruit.