Quantifying collective interactions in biomolecular phase separation

Abstract Biomolecular phase separation is an emerging theme for protein assembly and cellular organisation. The collective forces driving such condensation, however, remain challenging to characterise. Here we show that tracking the dilute phase concentration of only one component suffices to quanti...

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Main Authors: Hannes Ausserwöger, Ella de Csilléry, Daoyuan Qian, Georg Krainer, Timothy J. Welsh, Tomas Sneideris, Titus M. Franzmann, Seema Qamar, Nadia A. Erkamp, Jonathon Nixon-Abell, Mrityunjoy Kar, Peter St George-Hyslop, Anthony A. Hyman, Simon Alberti, Rohit V. Pappu, Tuomas P. J. Knowles
Format: Article
Language:English
Published: Nature Portfolio 2025-08-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-62437-y
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Summary:Abstract Biomolecular phase separation is an emerging theme for protein assembly and cellular organisation. The collective forces driving such condensation, however, remain challenging to characterise. Here we show that tracking the dilute phase concentration of only one component suffices to quantify composition and energetics of multicomponent condensates. Applying this assay to several disease- and stress-related proteins, we find that monovalent ions can either deplete from or enrich within the dense phase in a context-dependent manner. By analysing the effect of the widely used modulator 1,6-hexanediol, we find that the compound inhibits phase separation by acting as a solvation agent that expands polypeptide chains. Extending the strategy to in cellulo data, we even quantify the relative energetic contributions of individual proteins within complex condensates. Together, our approach provides a generic and broadly applicable tool for dissecting the forces governing biomolecular condensation and guiding the rational modulation of condensate behaviour.
ISSN:2041-1723