Double Membrane System Research Articles

Long-Range Optomechanical Interactions in SiN Membrane Arrays

Optomechanical systems using a membrane-in-the-middle configuration can exhibit a long-range type of interaction similar to how atoms show collective motion in an optical potential. Photons bounce back and forth inside a high-finesse Fabry-Pérot cavity and mediate the interaction between multiple membranes over a significant distance compared to the wavelength. Recently, it has been demonstrated that off-resonant coupling between light and the intermembrane cavity can lead to coherent mechanical noise cancellation. On-resonance coupling of light with both the Fabry-Pérot and intermembrane cavities, predicted to enhance the single-photon optomechanical coupling, have to date not been experimentally demonstrated, however. In our experiment, a double-membrane system inside a Fabry-Pérot cavity resonantly enhances the cavity field, resulting in a stronger optomechanical coupling strength from the increased radiation pressure. The resonance condition is first identified by analyzing the slope of the dispersion relation. Then, the optomechanical coupling is determined at various chip positions over one wavelength range. The optimum coupling conditions are obtained and enhancement is demonstrated for double-membrane arrays with three different reflectivites, reaching nearly fourfold enhancement for the collective motion of R=65% double membranes. The cavity losses at the optimum coupling are also characterized and the potential of reaching the single-photon strong coupling regime is discussed. Published by the American Physical Society 2025

Fission of double-membrane tubes under tension

The division of a cellular compartment culminates with the scission of a highly constricted membrane neck. Scission requires lipid rearrangements, topology changes, and transient formation of nonbilayer intermediate structures driven by curvature stress. Often, a side effect of this stress is pore-formation, which may lead to content leakage and thus breaching of the membrane barrier function. In single-membrane systems, leakage is avoided through the formation of a hemifusion (HF) intermediate, whose structure is still a subject of debate. The consequences of curvature stress have not been explored in double-membrane systems, such as the mitochondrion. Here, we combine experimental and theoretical approaches to study neck constriction and scission driven by tension in biomimetic lipid systems, namely single- and double-membrane nanotubes (sNTs and dNTs), respectively. In sNTs, constriction by high tension gives rise to a metastable HF intermediate (seen as stalk or worm-like micelle), whereas poration is universally slower in a simple neck. In dNTs, high membrane tension causes sequential rupture of each membrane. In contrast, low tension leads to the HF of both membranes, which may lead to a leaky fusion pathway, or may progress to further fusion of the two membranes along a number of transformation pathways. These findings provide a new mechanistic basis for fundamental cellular processes.

Membrane fission via transmembrane contact

Division of intracellular organelles often correlates with additional membrane wrapping, e.g., by the endoplasmic reticulum or the outer mitochondrial membrane. Such wrapping plays a vital role in proteome and lipidome organization. However, how an extra membrane impacts the mechanics of the division has not been investigated. Here we combine fluorescence and cryo-electron microscopy experiments with self-consistent field theory to explore the stress-induced instabilities imposed by membrane wrapping in a simple double-membrane tubular system. We find that, at physiologically relevant conditions, the outer membrane facilitates an alternative pathway for the inner-tube fission through the formation of a transient contact (hemi-fusion) between both membranes. A detailed molecular theory of the fission pathways in the double membrane system reveals the topological complexity of the process, resulting both in leaky and leakless intermediates, with energies and topologies predicting physiological events.

Use of Super-Resolution Live Cell Imaging to Distinguish Endoplasmic Reticulum: Nuclear Envelope Subcellular Localization.

A distinguishing feature of eukaryotes is the presence of a nuclear envelope (NE) and endomembrane system. The NE is a double-membrane system that surrounds chromatin and is continuous with the endoplasmic reticulum (ER). This interface is crucial in various processes such as calcium signaling and ER-associated degradation. The outer nuclear membrane and ER share a multitude of proteins although some are only functional in one domain, whereas the inner nuclear membrane has its own unique proteome. Until recently, it was not possible to distinguish between the inner and outer nuclear membranes as well as perinuclear ER using light microscopy - only electron microscopy was suitable for this. Now, however, using super-resolution live cell imaging, this can be achieved while still observing protein and membrane dynamics in real time. The protocols described here will allow researchers to determine subcellular localization of potential NE/ER proteins in live plant cells, helping to gain new insights into protein functionality.

The Role of the Gravitational Field in Generating Electric Potentials in a Double-Membrane System for Concentration Polarization Conditions.

Electric potentials referred to as the gravielectric effect (∆ΨS) are generated in a double-membrane system containing identical polymer membranes set in horizontal planes and separating non-homogenous electrolyte solutions. The gravielectric effect depends on the concentration and composition of the solutions and is formed due to the gravitational field breaking the symmetry of membrane complexes/concentration boundary layers formed under concentration polarization conditions. As a part of the Kedem-Katchalsky formalism, a model of ion transport was developed, containing the transport parameters of membranes and solutions and taking into account hydrodynamic (convective) instabilities. The transition from non-convective to convective or vice versa can be controlled by a dimensionless concentration polarization factor or concentration Rayleigh number. Using the original measuring set, the time dependence of the membrane potentials was investigated. For steady states, the ∆ΨS was calculated and then the concentration characteristics of this effect were determined for aqueous solutions of NaCl and ethanol. The results obtained from the calculations based on the mathematical model of the gravitational effect are consistent with the experimental results within a 7% error range. It has been shown that a positive or negative gravielectric effect appeared when a density of the solution in the inter-membrane compartment was higher or lower than the density in the outer compartments. The values of the ∆ΨS were in a range from 0 to 27 mV. It was found that, the lower the concentration of solutions in the outer compartments of the two-membrane system (C0), for the same values of Cm/C0, the higher the ∆ΨS, which indicates control properties of the double-membrane system. The considered two-membrane electrochemical system is a source of electromotive force and functions as an electrochemical gravireceptor.

Application of different treatment systems for boron removal from industrial wastewater with extremely high boron content

The aim of the study is purification and environmental management of boron containing industrial wastewater wastewaters comparing the efficiency of chemical precipitation and membrane filtration systems on the removal of boron at high concentrations from wastewater coming from boric acid production process and ground water. In order to eliminate adverse effects and find optimum treatment system, chemical precipitation, electrocoagulation and reverse osmosis processes have been studied. Different chemical precipitants such as poly aluminum chloride (PACl), lime, calcium chloride (CaCl2) and barium chloride (BaCl2) and hydrogen peroxide as oxidant were used. When solution pH was adjusted at 10, boron removal increased to 99.5 %. With application of electrocoagulation, 9 % of SO42-, 80 % of Si and 45 % of B were removed from spring water. At membrane-system, single and double pass BWRO/SWRO membrane system operations were carried out in order. Boron-removal increased 98 % by increasing pH up to 10, and DP-SWRO membranes system at 10-pH was found to be the best system. Obtaining a sufficient result for boron removal from spring water, a hybrid membrane process which contains an electrocoagulation process as a pre-treatment method and a double pass RO membrane system has been developed as an attractive technology.

Equivalence of Charge Imbalance and External Electric Fields during Free Energy Calculations of Membrane Electroporation.

Electric fields across lipid membranes play important roles in physiology, medicine, and biotechnology, rationalizing the wide interest in modeling transmembrane potentials in molecular dynamics simulations. Transmembrane potentials have been implemented with external electric fields or by imposing charge imbalance between the two water compartments of a stacked double-membrane system. We compare the two methods in the context of membrane electroporation, which involves a large change of membrane structure and capacitance. We show that, given that Ewald electrostatics are defined with tinfoil boundary conditions, the two methods lead to (i) identical potentials of mean force (PMFs) of pore formation and expansion at various potentials, demonstrating that the two methods impose equivalent driving forces for large-scale transitions at membranes, and (ii) to identical polarization of water within thin water wires or open pores, suggesting that the two methods furthermore impose equivalent local electric fields. Without tinfoil boundary conditions, effects from external fields on pore formation are spuriously suppressed or even removed. Together, our study shows that both methods, external fields and charge imbalance, are well suitable for studying large-scale transitions of lipid membranes that involve changes of membrane capacitance. However, using charge imbalance is technically more challenging for maintaining a constant transmembrane potential since it requires updating of the charge imbalance as the membrane capacitance changes.

NucEnvDB: A Database of Nuclear Envelope Proteins and Their Interactions.

The nuclear envelope (NE) is a double-membrane system surrounding the nucleus of eukaryotic cells. A large number of proteins are localized in the NE, performing a wide variety of functions, from the bidirectional exchange of molecules between the cytoplasm and the nucleus to chromatin tethering, genome organization, regulation of signaling cascades, and many others. Despite its importance, several aspects of the NE, including its protein-protein interactions, remain understudied. In this work, we present NucEnvDB, a publicly available database of NE proteins and their interactions. Each database entry contains useful annotation including a description of its position in the NE, its interactions with other proteins, and cross-references to major biological repositories. In addition, the database provides users with a number of visualization and analysis tools, including the ability to construct and visualize protein-protein interaction networks and perform functional enrichment analysis for clusters of NE proteins and their interaction partners. The capabilities of NucEnvDB and its analysis tools are showcased by two informative case studies, exploring protein-protein interactions in Hutchinson-Gilford progeria and during SARS-CoV-2 infection at the level of the nuclear envelope.

Modelling of an autonomous Nav1.5 channel system as a part of in silico pharmacology study.

A homology model of Nav1.5, based mainly on the crystal structures of Nav1.2/1.5 was built, optimized and successfully inserted into the membrane bilayer. We applied steered and free MD simulation protocols for the visualization of the mechanism of Nav1.5 activation. We constrained dihedrals of S4 trigger to introduce a structural tension with further rearrangement and movement of secondary structure elements. From these, we observed an intracellular gate opening and movement of the Lys1419 residue caused by a gradual displacement of the distal S6 α-helix with the extended S4 3-10 helix of voltage-sensing domains (VSD). A construction containing the Lys1419 residue in P-loop also changed its position due to the extension of this helix and subsequent induction of the pore-forming helixes motion. From this point, a double membrane system was generated, implying a free of ligand Nav1.5 protein and on the opposite side its copy containing a docked bupivacaine molecule inside the pore channel. The system can be used for the design of selective inhibitors against the Nav1.7 channel, instead of mixed effect on both channels.

Unique Properties of Tightly Docked Membranes Prior to their Fusion

Membrane merger through fusion is essential in myriad of processes including neurotransmission and viral infection. Although there are various classes of fusion proteins, they are believed to catalyze membrane fusion through a common pathway. Despite the importance of understanding of this process, the exact mechanisms underlying steps immediately before the first bilayer disruption remain uncharacterized. Here, we characterized mechanism leading to a change in lipid arrangement in membranes arrested in a state that precedes hemifusion. Such tightly adhering membranes with a distance of <1 nm are known from cryo-electron microscopy as fusion intermediates of trafficking organelles, mitochondria, or during cell entry by enveloped viruses. With the use of an in vitro reconstitution system consisting of lipid vesicles with SNARE proteins, we demonstrate formation of a metastable, divalent cation-independent, and protein-free membrane-membrane interface characterized by increased thickness. Furthermore, we confirm that structural rearrangements in the membranes are arising due to membrane intrinsic properties by performing all-atom molecular dynamics simulations of double-membrane systems at varying distances. In turn, we would like to propose that increase in membrane thickness arises by dehydration-driven lipid headgroup tilting leading to lateral area shrinkage that results in increased bilayer thickness visible in experiments.

Computational Investigation into the Effect of SS-31 on Membrane Ion Distributions, Pore Formation and Ion Leakage in the Presence of Transmembrane Potentials

Mitochondrial dysfunction encompasses a broad array of disorders related to aging and neurodegeneration, and often presents with altered membrane potentials and overproduction of reactive oxygen species (ROS). Szeto-Schiller (SS) peptides comprise a family of synthetic, amphipathic tetrapeptides with demonstrated efficacy against many mitochondrial disorders, and which accumulate strongly in mitochondria and bind to lipid bilayers. However, little is known regarding how SS peptides interact with lipid bilayers in the presence of a transmembrane potential. In this study, we analyzed the interactions of lead compound SS-31 (Elamipretide) with model membranes through molecular dynamics (MD) simulations. We performed equilibrium MD in single-bilayer systems to compare with experimental findings which showed that the peptide's ability to modulate membrane surface electrostatics is an important aspect of SS-31's mechanism of action. We went onto study how SS-31 influences membrane pore formation and ion leakage in the presence of transmembrane potentials. These simulations employed double-membrane systems and several computational techniques including external electric fields, explicit ion imbalances and the GROMACS computational electrophysiology module. We found that in the presence of a high membrane potential, SS-31's presence doubled the likelihood of transmembrane ion leakage compared to membrane-only systems. Most notably, in a seeming compensatory fashion, SS-31's interaction promoted anion leakage but inhibited cation leakage which ultimately preserved the cation gradient and membrane potential after leakage events. We propose a mechanism by which SS-31's mitoprotective properties could hinge on relieving dysfunctional hyperpolarization, and thereby halting damage from excessive ROS production yet still preserving the proton gradient needed for ATP production. This study could help elucidate how SS peptides correct this clinically-important complication of mitochondrial dysfunction and assist in enhancing the efficacy of future compound variants.

Simulation of S-Entropy Production during the Transport of Non-Electrolyte Solutions in the Double-Membrane System.

Using the classical Kedem–Katchalsky’ membrane transport theory, a mathematical model was developed and the original concentration volume flux (Jv), solute flux (Js) characteristics, and S-entropy production by Jv, and by Js in a double-membrane system were simulated. In this system, M1 and Mr membranes separated the l, m, and r compartments containing homogeneous solutions of one non-electrolytic substance. The compartment m consists of the infinitesimal layer of solution and its volume fulfills the condition Vm → 0. The volume of compartments l and r fulfills the condition Vl = Vr → ∞. At the initial moment, the concentrations of the solution in the cell satisfy the condition Cl < Cm < Cr. Based on this model, for fixed values of transport parameters of membranes (i.e., the reflection (σl, σr), hydraulic permeability (Lpl, Lpr), and solute permeability (ωl, ωr) coefficients), the original dependencies Cm = f(Cl − Cr), Jv = f(Cl − Cr), Js = f(Cl − Cr), = f(Cl − Cr), = f(Cl − Cr), Rv = f(Cl − Cr), and Rs = f(Cl − Cr) were calculated. Each of the obtained features was specially arranged as a pair of parabola, hyperbola, or other complex curves.

A Method of Human Eye Parameter Measurement Based on Laser Self-Mixing Interference

The eye is the most important sensory organ of the human body, and the cornea and anterior chamber are very important components In surgeries like vision correction, it is necessary to know the accurate corneal thickness and anterior chamber depth of human eyes. Their accurate measurement cannot only provide a reference for doctors, but also provide a strong guarantee of surgical results. In order to achieve high-precision measurements of human corneal thickness and anterior chamber depth, we develop an equivalent human-eye optical system and a measurement method for the thickness. This method designs an equivalent double-membrane system to simplify the physiological structure of the cornea and anterior chamber of the human eye and irradiate the double-membrane system through laser. The reflected light returns to the laser cavity and the original light self-mixing interference, and a photodetector collects and processes the interference signals to obtain the corneal thickness and anterior chamber depth. The correctness of the theoretical calculations is verified by the experimental system, and the measurement accuracy can reach the micrometer level.

Structural Determinants of Lipid Membrane Thickening at Close Distances

Membrane fusion and fission are key processes underlying, e.g., synaptic transmission. For a quantitative characterization of these processes at the molecular level, understanding the physics of lipid bilayer interactions at close distances is essential. We have observed increased membrane thickness for early membrane fusion intermediates in SNARE-mediated membrane fusion, resembling similar effects observed previously for multilamellar-membrane stacks. To reveal the underlying structural changes and molecular causes, we have carried out all-atom molecular dynamics simulations of double-membrane systems at decreasing mutual distances. Indeed, for distances below 1.5nm, thickening is seen also in the simulations, although to a smaller extent (11%). We also observed a decreased area per lipid and increased lipid chain order parameters, strongly reduced translational and rotational entropy for water molecules between the membranes, as well as markedly slowed and tightly correlated lateral diffusion of the inwards-facing lipid molecules. Particularly for close distances, the dipolar lipid head groups become increasingly tilted, which we identified as the main determinant for the observed thickening.

Ultrastructural modeling of small angle scattering from photosynthetic membranes

The last decade has seen a range of studies using non-invasive neutron and X-ray techniques to probe the ultrastructure of a variety of photosynthetic membrane systems. A common denominator in this work is the lack of an explicitly formulated underlying structural model, ultimately leading to ambiguity in the data interpretation. Here we formulate and implement a full mathematical model of the scattering from a stacked double bilayer membrane system taking instrumental resolution and polydispersity into account. We validate our model by direct simulation of scattering patterns from 3D structural models. Most importantly, we demonstrate that the full scattering curves from three structurally typical cyanobacterial thylakoid membrane systems measured in vivo can all be described within this framework. The model provides realistic estimates of key structural parameters in the thylakoid membrane, in particular the overall stacking distance and how this is divided between membranes, lumen and cytoplasmic liquid. Finally, from fitted scattering length densities it becomes clear that the protein content in the inner lumen has to be lower than in the outer cytoplasmic liquid and we extract the first quantitative measure of the luminal protein content in a living cyanobacteria.

The Quest for the Blueprint of the Nuclear Pore Complex.

During my postdoc interview in June of 1998, I asked Günter why he was moving more towards the nucleus in his latest studies. He said, "Well Joe, that's where everything starts." By the end of the interview, I accepted the postdoc. He had a way of making everything sound so cool. Günter's progression was natural, since the endoplasmic reticulum and the nucleus are the only organelles that share the same membrane. The nuclear envelope extends into a double membrane system with nuclear pore complexes embedded in the pore membrane openings. Even while writing this review, I remember Günter stressing; it is the nuclear pore complex. Just saying nuclear pore doesn't encompass the full magnitude of its significance. The nuclear pore complex is one of the largest collection of proteins that fit together for an overall function: transport. This review will cover the Blobel lab contributions in the quest for the blueprint of the nuclear pore complex from isolation of the nuclear envelope and nuclear lamin to the ring structures.

Unraveling Hidden Components of the Chloroplast Envelope Proteome: Opportunities and Limits of Better MS Sensitivity

The chloroplast is a major plant cell organelle that fulfills essential metabolic and biosynthetic functions. Located at the interface between the chloroplast and other cell compartments, the chloroplast envelope system is a strategic barrier controlling the exchange of ions, metabolites and proteins, thus regulating essential metabolic functions (synthesis of hormones precursors, amino acids, pigments, sugars, vitamins, lipids, nucleotides etc.) of the plant cell. However, unraveling the contents of the chloroplast envelope proteome remains a difficult challenge; many proteins constituting this functional double membrane system remain to be identified. Indeed, the envelope contains only 1% of the chloroplast proteins (i.e. 0.4% of the whole cell proteome). In other words, most envelope proteins are so rare at the cell, chloroplast, or even envelope level, that they remained undetectable using targeted MS studies. Cross-contamination of chloroplast subcompartments by each other and by other cell compartments during cell fractionation, impedes accurate localization of many envelope proteins. The aim of the present study was to take advantage of technologically improved MS sensitivity to better define the proteome of the chloroplast envelope (differentiate genuine envelope proteins from contaminants). This MS-based analysis relied on an enrichment factor that was calculated for each protein identified in purified envelope fractions as compared with the value obtained for the same protein in crude cell extracts. Using this approach, a total of 1269 proteins were detected in purified envelope fractions, of which, 462 could be assigned an envelope localization by combining MS-based spectral count analyses with manual annotation using data from the literature and prediction tools. Many of such proteins being previously unknown envelope components, these data constitute a new resource of significant value to the broader plant science community aiming to define principles and molecular mechanisms controlling fundamental aspects of plastid biogenesis and functions.

Translocase of the Outer Mitochondrial Membrane 40 Is Required for Mitochondrial Biogenesis and Embryo Development in Arabidopsis.

In eukaryotes, mitochondrion is an essential organelle which is surrounded by a double membrane system, including the outer membrane, intermembrane space and the inner membrane. The translocase of the outer mitochondrial membrane (TOM) complex has attracted enormous interest for its role in importing the preprotein from the cytoplasm into the mitochondrion. However, little is understood about the potential biological function of the TOM complex in Arabidopsis. The aim of the present study was to investigate how AtTOM40, a gene encoding the core subunit of the TOM complex, works in Arabidopsis. As a result, we found that lack of AtTOM40 disturbed embryo development and its pattern formation after the globular embryo stage, and finally caused albino ovules and seed abortion at the ratio of a quarter in the homozygous tom40 plants. Further investigation demonstrated that AtTOM40 is wildly expressed in different tissues, especially in cotyledons primordium during Arabidopsis embryogenesis. Moreover, we confirmed that the encoded protein AtTOM40 is localized in mitochondrion, and the observation of the ultrastructure revealed that mitochondrion biogenesis was impaired in tom40-1 embryo cells. Quantitative real-time PCR was utilized to determine the expression of genes encoding outer mitochondrial membrane proteins in the homozygous tom40-1 mutant embryos, including the genes known to be involved in import, assembly and transport of mitochondrial proteins, and the results demonstrated that most of the gene expressions were abnormal. Similarly, the expression of genes relevant to embryo development and pattern formation, such as SAM (shoot apical meristem), cotyledon, vascular primordium and hypophysis, was also affected in homozygous tom40-1 mutant embryos. Taken together, we draw the conclusion that the AtTOM40 gene is essential for the normal structure of the mitochondrion, and participates in early embryo development and pattern formation through maintaining the biogenesis of mitochondria. The findings of this study may provide new insight into the biological function of the TOM40 subunit in higher plants.

Mechanical and Electric Control of Photonic Modes in Random Dielectrics.

Random dielectrics defines a class of non-absorbing materials where the index of refraction is randomly arranged in space. Whenever the transport mean free path is sufficiently small, light can be confined in modes with very small volume. Random photonic modes have been investigated for their basic physical insights, such as Anderson localization, and recently several applications have been envisioned in the field of renewable energies, telecommunications, and quantum electrodynamics. An advantage for optoelectronics and quantum source integration offered by random systems is their high density of photonic modes, which span a large range of spectral resonances and spatial distributions, thus increasing the probability to match randomly distributed emitters. Conversely, the main disadvantage is the lack of deterministic engineering of one or more of the many random photonic modes achieved. This issue is solved by demonstrating the capability to electrically and mechanically control the random modes at telecom wavelengths in a 2D double membrane system. Very large and reversible mode tuning (up to 50 nm), both toward shorter or longer wavelength, is obtained for random modes with modal volumes of the order of few tens of (λ/n)3 .

Double-Membrane Sampling System in a Mass Spectrometer for the Study of Exhaled Air

A double-membrane sample-introduction system is described that enables mass spectrometric determination of volatile organic compounds in air, while ensuring their detection limits are as low as a few fractions of ppb of sample composition in real time with a response delay from a few seconds to a few dozens of seconds. It includes two membrane interfaces separated by a volume, the gas pressure in which is maintained through a channel with an adjustable evacuation cross-section. The results of numerical simulation and experimental testing of the system are provided. A technique for selecting and calculating system parameters is described. The possibility of using this system for diagnosing diseases by trace amounts of biomarkers in exhaled air is discussed.