Increasing the Compositional Heterogeneity of Single-Chain Amphiphile Membranes Supported by Coacervate Cores Alters Stability and Properties of the Hybrid Protocells.

Coacervate droplets and lipid vesicles are two classes of self-assembled compartments that have been proposed as protocell models. Hybrid protocells, in which a coacervate core is surrounded by a lipid membrane, can integrate the advantages of both protocell systems while overcoming their limitations. Although hybrid protocell membranes have been produced with a variety of diacyl phospholipids related to modern biology and some single-chain amphiphiles inspired by prebiotic scenarios, little is known about how mixtures of single-chain amphiphiles impact hybrid protocell membrane formation and properties. Given the plausible diversity of amphiphiles in the prebiotic milieu, the resulting membranes would have inherently incorporated multiple lipids of different types, potentially altering the properties and viability of hybrid protocells in their environment. Here, we systematically increased the compositional heterogeneity of hybrid protocell membranes by using different prebiotically relevant single-chain amphiphiles of varying head groups and alkyl chain lengths. These membranes were assembled around model coacervate droplets generated from poly(allylamine hydrochloride) and adenosine diphosphate, and the effect of heterogeneity on membrane properties and stability was evaluated. Compared to protocells with homogeneous membranes, those with heterogeneous amphiphile membranes exhibited higher yields, smaller sizes, and greater subcompartment formation. Also, they showed increased membrane order, retained similar lateral lipid diffusion, and showed population-level variability in permeability to small anionic molecules. Notably, heterogeneous membranes showed enhanced structural stability under acidic conditions, retaining key properties like size and subcompartment heterogeneity, thereby broadening the pH range over which hybrid protocells remain intact. These findings suggest that amphiphile diversity not only would have influenced the structural properties of hybrid protocells but also created diversity within the protocell population and enhanced their robustness, thereby playing a crucial role in protocell evolution on early Earth.