Dissecting membrane interfacial cellular processes: an in vitro reconstitution approach

Cellular membrane interfacial processes are highly complex phenomena that are challenging to explore in vivo, often limiting their mechanistic understanding. The plethora of participating biological entities and their unprecedented complex interactions might also mask their core guiding principles. In vitro reconstitution experiments with a reductionist bottom-up approach attempt to dissect the biological complexity by recreating biological reactions in their simplest form. This review focuses on the cellular membrane interfacial processes crucial for cellular transport, neurodegeneration, and host-pathogen interactions. We discuss how reconstitution methodologies have played a crucial role in addressing many of the fundamental questions dealing with the mechanisms of clathrin-dependent or independent endocytosis and the Endosomal Sorting Complex Required for Transport (ESCRT) pathway. In addition, we have discussed how reconstitution-based approaches helped explore membrane-mediated alterations in the onset of neurodegenerative diseases and host-pathogen interactions. Importantly, the reconstitution approach, though predominantly applied but not limited to addressing questions on cellular membrane shaping, is gradually gaining attention across multiple areas of cell biology.