Dynamic principles of concentration buffering through liquid-liquid phase separation.

Abstract

Living systems must maintain robust biochemical function despite fluctuations that span a wide range of timescales. Biomolecular condensates formed by liquid-liquid phase separation (LLPS) have been shown to buffer concentration fluctuations, but the principles governing their dynamic regulation remain unclear. We address this by probing the response of LLPS to oscillatory perturbations that mimic fluctuations across different timescales, establishing the first systematic frequency-domain analysis of concentration buffering by condensates. We find that condensates act as frequency-selective filters: The perturbed dilute phase behaves as a high-pass filter, while the dense phase attenuates both low- and high-frequency perturbations. We establish quantitative links between LLPS parameters, including interaction strength, droplet size, and interphase exchange rates, and the timescale range over which condensates effectively buffer concentration fluctuations. These findings establish fundamental dynamical limits of concentration buffering by LLPS, with implications for how cells may use LLPS to adapt to fluctuating environments and for the design of synthetic condensates with programmable control properties.