Membrane Interfacial Potential Governs Surface Condensation and Fibrillation of α-Synuclein in Neurons

Abstract Biomolecular condensates formed via liquid-liquid phase separation (LLPS) are essential for cellular organization. α-Synuclein, an amyloidogenic protein linked to Parkinson’s Disease (PD), undergoes phase separation at high concentrations, but the influence of lipid membranes on this process remains unclear. Here, combining in vitro reconstitution, cell biology, and simulations, we show that membranous interfaces promote α-Synuclein condensation at physiologically relevant sub-critical concentrations (∼10 nM) without crowding agents. Notably, condensation occurs only on membranes with a specific stoichiometry of lipids, underscoring the role of interfacial potential. These condensates serve as nucleation sites for fibril formation, leading to membrane deformation and rupture. A lattice gas model reveals this behavior as a prewetting-like transition, where an attractive membrane induces local phase separation below the bulk saturation concentration. Indeed altering interfacial potential by lipid composition and membrane depolarization not only drastically changes α-Synuclein puncta size and number but also triggers their release from neurons. These findings reveal the crucial role of lipid membrane interfaces in regulating α-Synuclein condensation, aggregation and release, shedding light on a potential mechanism of their cell-to-cell propagation during neurodegeneration. Competing Interest Statement The authors have declared no competing interest. Footnotes The first author's details have been corrected on the title page of the manuscript.