Feedback-induced phase separation of hollow condensates to create biomimetic membraneless compartments

Abstract

Intracellular macromolecules have the ability to form membraneless compartments, such as vacuoles and hollow condensates, through liquid–liquid phase separation (LLPS) in order to adapt to changes in their environment. The development of artificial non-homogeneous compartments, such as multiphase hollow or multicavity condensates, has gained significant attention due to their potential to uncover the mechanisms underlying the formation of artificial condensates and biomolecular condensates. However, the complexity of design and construction has hindered progress, particularly in creating dynamic non-homogeneous compartments. In this study, we present a dynamic membraneless compartment using peptide–oligonucleotide conjugates derived from short elastin-like polypeptides (sELP–ONs), which undergo pH-mediated phase transition. Below pH 8.8, the microcompartment exists as microdroplets that transform into non-homogeneous hollow condensates above pH 8.8. Notably, these hollow condensates retain liquid properties and high molecular ordering, and effectively sequester guest molecules with a hollow condensed layer. Furthermore, our sELP–ON microcompartments exhibit a feedback-induced phase transition in response to environmental pH fluctuations generated by complex enzymatic reactions mimicking cellular metabolism, providing a novel dynamic model for creating biomimetic membraneless compartments.