The proliferation of life on earth is based on the ability of single cells to divide into two daughter cells. During cell division, the plasma membrane undergoes a series of morphological transformations which ultimately lead to membrane fission. Here, we show that analogous remodeling processes can be induced by low densities of proteins bound to the membranes of cell-sized lipid vesicles. Using His-tagged fluorescent proteins, we are able to precisely control the spontaneous curvature of the vesicle membranes. By fine-tuning this curvature, we obtain dumbbell-shaped vesicles with closed membrane necks as well as neck fission and complete vesicle division. Our results demonstrate that the spontaneous curvature generates constriction forces around the membrane necks and that these forces can easily cover the force range found in vivo. Our approach involves only one species of membrane-bound proteins at low densities, thereby providing a simple and extendible module for bottom-up synthetic biology.

Simple sugars such as glucose and sucrose are ubiquitous in all organisms. One remarkable property of these small solutes is their ability to protect biomembranes against dehydration damage. This property, which reflects the underlying sugar–lipid interactions, has been intensely studied for lipid bilayers interacting with a single sugar at low hydration. Here, we use giant vesicles to investigate fully hydrated lipid membranes in contact with two sugars, glucose and sucrose. The vesicles were osmotically balanced, with the same total sugar concentration in the interior and exterior aqueous solutions. However, the two solutions differed in their composition: the interior solution contained only sucrose whereas the exterior one contained primarily glucose. This sugar asymmetry generated a striking variety of multispherical or “multi-balloon” vesicle shapes. Each multisphere involved only a single membrane that formed several spherical segments, which were connected by narrow, hourglass-shaped membrane necks. These morphologies revealed that the sugar–lipid interactions generated a significant spontaneous curvature with a magnitude of about 1 μm⁻¹. Such a spontaneous curvature can be generated both by depletion and by adsorption layers of the sugar molecules arising from effectively repulsive and attractive sugar–lipid interactions. All multispherical shapes are stable over a wide range of parameters, with a substantial overlap between the different stability regimes, reflecting the rugged free energy landscape in shape space. One challenge for future studies is to identify pathways within this landscape that allow us to open and close the membrane necks of these shapes in a controlled and reliable manner. We will then be able to apply these multispheres as metamorphic chambers for chemical reactions and nanoparticle growth.

Giant unilamellar vesicles (GUVs) provide a direct connection between the nano- and the microregime. On the one hand, these vesicles represent biomimetic compartments with linear dimensions of many micrometers. On the other hand, the vesicle walls are provided by single molecular bilayers that have a thickness of a few nanometers and respond sensitively to molecular interactions with small solutes, biopolymers, and nanoparticles. These nanoscopic responses are amplified by the GUVs and can then be studied on much larger scales. Therefore, GUVs are increasingly used as a versatile research tool for basic membrane science, bioengineering, and synthetic biology. Conventional GUVs have one major drawback, however: they have only a limited capability to cope with external perturbations such as osmotic inflation, adhesion, or micropipette aspiration that tend to rupture the membranes. In contrast, cell membranes tolerate the same kinds of mechanical perturbations without rupture because the latter membranes are coupled to reservoirs of membrane area. Here, we introduce GUVs with membrane nanotubes as model systems that include such area reservoirs. To demonstrate the increased robustness of these tubulated vesicles, we use micropipette aspiration and changes in the osmotic conditions applied to phospholipid membranes doped with the glycolipid GM1. A quantitative comparison between theory and experiment reveals that the response of the GUVs is governed by the membranes’ spontaneous tension, a curvature-elastic material parameter that describes the bilayer asymmetry on the nanoscale. Because of their increased robustness, GUVs with nanotubes represent improved research tools for membrane science, in general, with potential applications as storage and delivery systems and as cell-like microcompartments in bioengineering, pharmacology, and synthetic biology.

A microfluidic method to measure membrane permeability by trapping giant vesicles and performing quick solution exchange together with confocal microscopy.