Lipid coated protein condensates as stable protocells with selective uptake abilities for biomolecules

Abstract

To create cell-like synthetic systems, spatial confinement that is stable against environmental changes and selective uptake of diverse biomolecules into these compartments are key initial conditions. However, fabrication of protocells with these two features has been extremely difficult. Here, we used fully protein-based liquid condensates and a lipid coating on these condensates to construct highly stable protocells with an uptake ability for outside biomolecules. Condensates with an extremely high density of 6His-tagged proteins were coated with Ni(II)-NTA(nitrilotriacetic acid)-modified lipids. High condensate rigidity and specific 6His-Ni-NTA interactions enabled the formation of lipid–protein protocells, which are stable even after centrifugations. In addition, immobile lipid coatings on condensates were permeable to outside biomolecules. When binding modules were fused into condensate-forming proteins, the resulting functionalized condensate-protocells could strongly and selectively uptake various outside proteins through specific protein interactions.