Electrophysiological Characterization of Monoolein-FattyAcid Bilayers

Understanding the evolution of protocells, primitive compartments that distinguish self from nonself, is crucial for exploring the origin of life. Fatty acids and monoglycerides have been proposed as key components of protocell membranes due to their ability to self-assemble into bilayers and vesicles capable of nutrient exchange. In this study, we investigate the electrophysiological properties of planar bilayers composed of monoglyceride and fatty acid mixtures, using a droplet interface bilayer system. Three fatty acids with varying hydrocarbon chain lengthsoleic acid (C18), palmitoleic acid (C16), and myristoleic acid (C14)in combination with monoolein (C18) are examined to evaluate the influence of chain length and composition on bilayer stability, thickness, and ion permeability. The results show that pure monoolein bilayers exhibit enhanced ion permeability compared to phospholipid bilayers, which are characteristic of modern cellular membranes. Furthermore, the incorporation of fatty acids into monoolein bilayers destabilizes the membrane structure and further increases ion permeability. We consider that this increased permeability is likely driven by three molecular characteristics. First, the wedge-like shape of monoolein may disrupt bilayer packing and induce transient pore formation. Second, the rapid flip-flop of fatty acids between bilayer leaflets likely facilitates ion transport. Third, the chain-length mismatch between monoolein and myristoleic acid further destabilizes the bilayer, promoting the formation of structural defects. These findings suggest that compositional motifs in monoglyceride-fatty acid bilayers may provide an alternative ion transport mechanism, such as the flip-flop of amphiphilic molecules, in early protocell membranes before the evolution of protein-based transporters.