Cation-Driven Modulation of Tau Condensates: Insights into Liquid–Liquid Phase Separation and Rheological Properties

The formation of biocondensates through liquid-liquid phase separation (LLPS) has emerged as a vital and ubiquitous phenomenon contributing to the spatiotemporal coordination of cellular processes. Additionally, dysregulation of LLPS is increasingly implicated as a previously overlooked driver of diseases. LLPS typically involves multivalent noncovalent interactions among biomolecules, yet the role of solvent molecules, particularly water, in this process has received increasing attention. Metal ions are essential for life and exist in varying concentrations within cells. Both the concentration and type of metal ions significantly influence the phase separation of biomolecules. Ions with different degrees of hydration can uniquely alter the structure of water, which, in turn, affects LLPS. In this study, we use hyperspectral imaging (HSI) analysis and optical tweezers to investigate the effects of cations with different degrees of hydration on solvent properties within Tau condensates, an intrinsically disordered protein involved in Alzheimer's disease. We first demonstrate that the environment within Tau droplets is more structured than the diluted phase. Then, we show that highly hydrated cations enhance phase separation, increase the proportion of restricted water within Tau droplets, and slow down their relaxation dynamics, suggesting a correlation between water structuring and rheological properties. By connecting solvent properties with the stability and dynamics of phase-separated droplets, this research provides insights into the molecular mechanisms governing LLPS and how environmental factors, such as metal ions and water structure, influence this process.