The cell envelope in Gram-negative bacteria comprises two distinct membranes with a cell wall between them. There has been a growing interest in understanding the mechanical adaptation of this cell ...envelope to the osmotic pressure (or turgor pressure), which is generated by the difference in the concentration of solutes between the cytoplasm and the external environment. However, it remains unexplored how the cell wall, the inner membrane (IM), and the outer membrane (OM) effectively protect the cell from this pressure by bearing the resulting surface tension, thus preventing the formation of inner membrane bulges, abnormal cell morphology, spheroplasts and cell lysis. In this study, we have used molecular dynamics (MD) simulations combined with experiments to resolve how and to what extent models of the IM, OM, and cell wall respond to changes in surface tension. We calculated the area compressibility modulus of all three components in simulations from tension-area isotherms. Experiments on monolayers mimicking individual leaflets of the IM and OM were also used to characterize their compressibility. While the membranes become softer as they expand, the cell wall exhibits significant strain stiffening at moderate to high tensions. We integrate these results into a model of the cell envelope in which the OM and cell wall share the tension at low turgor pressure (0.3 atm) but the tension in the cell wall dominates at high values (>1 atm).
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•The cell envelope of Gram-negative bacteria is composed of three layers: inner and outer membranes and the cell wall.•These layers work together to resist turgor pressure coming from within.•Simulations and experiments on each layer reveals their mechanical properties, which are combined in a model of their collective behavior.•The cell wall is predicted to exist in a highly strained state in the living cell.•Both the cell wall and outer membrane make significant contributions to resisting turgor pressure.
Mechanical properties of a lipid bilayer are parameters determined mainly for giant unilamellar vesicles (GUVs). It is not clear if values obtained on the GUV model can be directly translated to ...submicron large unilamellar vesicles (LUVs). This ambiguity is a major obstacle in exploring the effect of lipid bilayer mechanics on membrane associated processes and effectiveness of liposome-based targeted drug delivery systems. In presented work extrusion, which is a common method to prepare LUVs, was used to study liposomes preparation and stability upon exposure to mechanical stress. The effect of parameters of the extrusion process (temperature, membrane pore size, extrusion force and volumetric flux) on the properties of liposome suspension (average liposome size, polydispersity index and lipid recovery ratio) was determined for model liposomes composed of DPPC lipid. The state of the DPPC lipid bilayer depends on temperature, therefore, the effect of lipid bilayer mechanics on the extrusion process can be quantitated without altering membrane composition. The extrusion process was carried out with the automated extruder delivering quantitative data on the extrusion force and volumetric flux. Obtained results have been interpreted in terms of mechanical properties of the lipid bilayer. Determined mechanical properties of the lipid bilayer and its dependence on temperature are in good agreement with the literature results determined for GUVs. This shows that mechanical properties of the lipid bilayer does not depend on the liposome size in the range from 100 nm to hundreds of microns.
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•Using the quantitative extrusion method, the bending rigidity coefficient of the lipid bilayer can be measured.•Using the quantitative extrusion method, the viscosity of liposome suspension in confine space can be determined.•Liposome recalibration process depends on the lipid bilayer mechanics.
Small unilamellar vesicles (SUVs), ubiquitous in organisms, play key and active roles in various biological processes. Although the physical properties of the constituent lipid molecules (i.e., the ...acyl chain length and saturation) are known to affect the mechanical properties of SUVs and consequently regulate their biological behaviors and functions, the underlying mechanism remains elusive. Here, we combined theoretical modeling and experimental investigation to probe the mechanical behaviors of SUVs with different lipid compositions. The membrane bending rigidity of SUVs increased with increasing chain length and saturation, resulting in differences in the vesicle rigidity and deformable capacity. Furthermore, we tested the tumor delivery capacity of liposomes with low, intermediate, and high rigidity as typical models for SUVs. Interestingly, liposomes with intermediate rigidity exhibited better tumor extracellular matrix diffusion and multicellular spheroid (MCS) penetration and retention than that of their stiffer or softer counterparts, contributing to improved tumor suppression. Stiff SUVs had superior cellular internalization capacity but intermediate tumor delivery efficacy. Stimulated emission depletion microscopy directly showed that the optimal formulation was able to transform to a rod-like shape in MCSs, which stimulated fast transport in tumor tissues. In contrast, stiff liposomes hardly deformed, whereas soft liposomes changed their shape irregularly, which slowed their MCS penetration. Our findings introduce special perspectives from which to map the detailed mechanical properties of SUVs with different compositions, provide clues for understanding the biological functions of SUVs, and suggest that liposome mechanics may be a design parameter for enhancing drug delivery.
F1Fo-ATP synthase (ATP synthase) is a central membrane protein that synthetizes most of the ATP in the cell through a rotational movement driven by a proton gradient across the hosting membrane. In ...mitochondria, ATP synthases can form dimers through specific interactions between some subunits of the protein. The dimeric form of ATP synthase provides the protein with a spontaneous curvature that sustain their arrangement at the rim of the high-curvature edges of mitochondrial membrane (cristae). Also, a direct interaction with cardiolipin, a lipid present in the inner mitochondrial membrane, induces the dimerization of ATP synthase molecules along cristae. The deletion of those biochemical interactions abolishes the protein dimerization producing an altered mitochondrial function and morphology. Mechanically, membrane bending is one of the key deformation modes by which mitochondrial membranes can be shaped. In particular, bending rigidity and spontaneous curvature are important physical factors for membrane remodelling. Here, we discuss a complementary mechanism whereby the rotatory movement of the ATP synthase might modify the mechanical properties of lipid bilayers and contribute to the formation and regulation of the membrane invaginations.
•Large uncertainties exist for values of the bending modulus of membranes.•The effect of sugar is considered, including new X-ray data.•Effect of tilt on data analysis is evaluated for different ...experimental methods.•Estimates of values of the tilt modulus are discussed.
Values of the bending modulus KC are reviewed, and possible causes for the considerable differences are discussed. One possible cause is the use of glucose and sucrose in the classical micromechanical manipulation and shape analysis methods. New data, using the more recent low angle X-ray method, are presented that do not support an effect of glucose or sucrose on KC. Another possible cause is using an incomplete theory to interpret the data. Adding a tilt term to the theory clearly does not affect the value obtained from the shape analysis method. It is shown that a tilt term, using a value of the modulus Kθ indicated by simulations, theory, and estimated from order parameters obtained from NMR and from the wide angle X-ray method, should also not affect the value obtained using the micromechanical manipulation method, although it does require a small correction when determining the value of the area compressibility modulus KA. It is still being studied whether including a tilt term will significantly affect the values of KC obtained using low angle X-ray data. It remains unclear what causes the differences in the experimental values of KC for simple lipid bilayers.
Piezo1 is an ion channel that gates open when mechanical force is applied to a cell membrane, thus allowing cells to detect and respond to mechanical stimulation. Molecular structures of Piezo1 ...reveal a large ion channel with an unusually curved shape. This study analyzes how such a curved ion channel interacts energetically with the cell membrane. Through membrane mechanical calculations, we show that Piezo1 deforms the membrane shape outside the perimeter of the channel into a curved 'membrane footprint'. This membrane footprint amplifies the sensitivity of Piezo1 to changes in membrane tension, rendering it exquisitely responsive. We assert that the shape of the Piezo channel is an elegant example of molecular form evolved to optimize a specific function, in this case tension sensitivity. Furthermore, the predicted influence of the membrane footprint on Piezo gating is consistent with the demonstrated importance of membrane-cytoskeletal attachments to Piezo gating.
Membrane fusion is a ubiquitous process associated with a multitude of biological events. Although it has long been appreciated that membrane mechanics plays an important role in membrane fusion, the ...molecular interplay between mechanics and fusion has remained elusive. For example, although different lipids modulate membrane mechanics differently, depending on their composition, molar ratio, and complex interactions, differing lipid compositions may lead to similar mechanical properties. This raises the question of whether (i) the specific lipid composition or (ii) the average mesoscale mechanics of membranes acts as the determining factor for cellular function. Furthermore, little is known about the potential consequences of fusion on membrane disruption. Here, we use a combination of confocal microscopy, time-resolved imaging, and electroporation to shed light onto the underlying mechanical properties of membranes that regulate membrane fusion. Fusion efficiency follows a nearly universal behavior that depends on membrane fluidity parameters, such as membrane viscosity and bending rigidity, rather than on specific lipid composition. This helps explaining why the charged and fluid membranes of the inner leaflet of the plasma membrane are more fusogenic than their outer counterparts. Importantly, we show that physiological levels of cholesterol, a key component of biological membranes, has a mild effect on fusion but significantly enhances membrane mechanical stability against pore formation, suggesting that its high cellular levels buffer the membrane against disruption. The ability of membranes to efficiently fuse while preserving their integrity may have given evolutionary advantages to cells by enabling their function while preserving membrane stability.
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