The mechanical response of giant liposomes to compression between two parallel plates is investigated in the context of an artificial actin cortex adjacent to the inner leaflet of the bilayer. We found that nonlinear membrane theory neglecting the impact of bending sufficiently describes the mechanical response of liposomes consisting of fluid lipids to compression whereas the formation of an actin cortex or the use of gel-phase lipids generally leads to substantial stiffening of the shell. Giant vesicles are gently adsorbed on glassy surfaces and are compressed with tipless cantilevers using an atomic force microscope. Force-compression curves display a nonlinear response that allows us to determine the membrane tension σ0 and the area compressibility modulus K(A) by computing the contour of the vesicle as a function of the compression depth. The values for KA of fluid membranes correspond well to what is known from micropipet-suction experiments and computed from monitoring membrane undulations. The presence of a thick actin shell adjacent to the inner leaflet of the liposome membrane stiffens the system considerably, as mirrored in a significantly higher apparent area compressibility modulus.

Indentation of giant liposomes with a conical indenter is described by means of a tension-based membrane model. We found that nonlinear membrane theory neglecting the impact of bending sufficiently describes the mechanical response of liposomes to indentation as measured by atomic force microscopy. Giant vesicles are gently adsorbed on glassy surfaces via avidin-biotin linkages and indented centrally using an atomic force microscope equipped with conventional sharp tips mounted on top of an inverted microscope. Force indentation curves display a nonlinear response that allows to extract pre-stress of the bilayer T0 and the area compressibility modulus KA by computing the contour of the vesicle at a given force. The values for KA of fluid membranes correspond well to what is known from micropipet suction experiments and inferred from membrane undulation monitoring. Assembly of actin shells inside the liposome considerably stiffens the vesicles resulting in significantly larger area compressibility modules. The analysis can be easily extended to different indenter geometries with rotational symmetry.

The epsin N-terminal homology domain (ENTH) is a major player in clathrin-mediated endocytosis. To investigate the influence of initial membrane tension on ENTH binding and activity, we established a bilayer system based on adhered giant unilamellar vesicles (GUVs) to be able to control and adjust the membrane tension covering a broad regime. The shape of each individual adhered GUV as well as its adhesion area was monitored by spinning disc confocal laser microscopy. Control of in a range of 0.08-1.02 mN/m was achieved by altering the Mg 2؉ concentration in solution, which changes the surface adhesion energy per unit area of the GUVs. Specific binding of ENTH to phosphatidylinositol 4,5-bisphosphate leads to a substantial increase in adhesion area of the sessile GUV. At low tension (<0.1 mN/m) binding of ENTH can induce tubular structures, whereas at higher membrane tension the ENTH interaction deflates the sessile GUV and thereby increases the adhesion area. The increase in adhesion area is mainly attributed to a decrease in the area compressibility modulus K A . We propose that the insertion of the ENTH helix-0 into the membrane is largely responsible for the observed decrease in K A , which is supported by the observation that the mutant ENTH L6E shows a reduced increase in adhesion area. These results demonstrate that even in the absence of tubule formation, the area compressibility modulus and, as such, the bending rigidity of the membrane is considerably reduced upon ENTH binding. This renders membrane bending and tubule formation energetically less costly.Clathrin-mediated endocytosis is one of the key metabolic pathways for the uptake of macromolecules into eukaryotic cells (1-4). Driven by a chain of remodeling events and an elaborate set of proteins acting in an orchestrated manner, an almost flat patch of plasma membrane is transformed into a closed, cargo-containing vesicle. As plasma membrane shape transformation is associated with significant local bending of the membrane, the process is highly sensitive to lateral membrane tension. Plasma membrane tension originates from two primary sources; that is, hydrostatic pressure across the lipid bilayer and cytoskeleton-membrane adhesion (5). Depending on the cell type, plasma membrane tensions span a range of roughly 0.003-0.45 mN/m (5-8). Even though it had become clear already in the late 1990s that tension plays a role in exoand endocytosis (9, 10), only in recent years has significant evidence been accumulated that membrane tension is of utmost importance for processes that rely on membrane remodeling (11-14). Cells actively maintain and regulate their membrane tension and use it to control exo-and endocytosis (15). In K562 cells it has been reported that endocytosis is completely suppressed under hypoosmotic conditions (16). Generally, high lateral membrane tension suppresses membrane deformation as both stretching the lipid bilayer and opening bonds between the cytoskeleton and the plasma membrane requires a large amount of energy (17).One protein...

The large gap in time scales between membrane fusion occurring in biological systems during neurotransmitter release and fusion observed between model membranes has provoked speculations over a large number of possible factors that might explain this discrepancy. One possible reason is an elevated lateral membrane tension present in the presynaptic membrane. We investigated the tension-dependency of fusion using model membranes equipped with a minimal fusion machinery consisting of syntaxin 1, synaptobrevin and SNAP 25. Two different strategies were realized; one based on supported bilayers and the other one employing sessile giant liposomes. In the first approach, isolated patches of planar bilayers derived from giant unilamellar vesicles containing syntaxin 1 and preassembled SNAP 25 (ΔN-complex) were deposited on a dilatable PDMS sheet. In a second approach, lateral membrane tension was controlled through the adhesion of intact giant unilamellar vesicles on a functionalized surface. In both approaches fusion efficiency increases considerably with lateral tension and we identified a threshold tension of 3.4 mN m−1, at which the number of fusion events is increased substantially.