Dynamic and Diverse Coacervate Architectures by Controlled Demembranization

The dynamics of membranes are integral to regulating biological pathways in living systems, particularly in mediating intra- and extracellular communication between membraneless and membranized nano- and microcompartments. Mimicking these dynamics using biomimetic cell structures deepens our understanding of biologically driven processes, including morphological transformations, communication, and molecular sequestration within distinct environments (e.g., (membraneless) organelles, cytoplasm, cells, and the extracellular matrix). In this context, the demembranization of membranized coacervates represents a promising approach to endow them with additional functionalities and dynamic reconfiguration capabilities in response to external or biological stimuli. This versatility broadens their applicability in synthetic biology, systems biology, and biotechnology. Here, we present a strategy for controlled demembranization of membranized coacervate droplets. The membranized coacervates are created by coating membraneless coacervates with terpolymer-based nanoparticles to form a solid-like membrane. The addition of an anionic polysaccharide then triggers the demembranization process arising from electrostatic competition with the membrane components, resulting in polysaccharide-containing demembranized coacervate droplets. This membranization/demembranization process not only allows for the controlled structural reconfiguration of the coacervate entities but also varies their permeability toward (biological) (macro)molecules and nano- and microscale objects. Additionally, integrating an additional polymersome layer in this process facilitates the creation of bilayer and ″Janus-like″ membranized coacervates, advancing the development of coacervate protocells with hierarchical and asymmetric membrane structures. Our work highlights the control over both membranization and demembranization processes of coacervate protocells, establishing a platform for creating advanced protein-containing synthetic protocells with dynamic and diverse (membrane(less)) architectures.