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bioRxiv · 10.1101/2025.06.19.660548

Amphiphilic protein surfactants reduce the interfacial tension of biomolecular condensates

Favetta, B.·Wang, H.·Shi, Z.·Schuster, B. S.
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Abstract

Biomolecular condensates are protein-dense regions in cells that often arise from liquid-liquid phase separation. Interfacial tension is a key determinant of biomolecular condensate behavior, influencing condensate size and interactions with intracellular structures. Certain proteins and RNAs are known to selectively localize to the interface of condensates, where they can regulate condensate function in cells. Previously, we designed amphiphilic proteins that preferentially adsorb to the surface of condensates. These proteins contain one phase-separating domain (RGG) and one non-phase-separating domain (MBP or GST). Here, we demonstrate through direct quantification that these amphiphilic proteins act as surfactants, reducing the interfacial tension of RGG-RGG condensates from [~]260 {micro}N/m to [~]100 {micro}N/m in a concentration-dependent manner. Notably, the GST-based surfactant protein exhibits a 10-fold greater efficacy in lowering interfacial tension compared to the MBP-based surfactant. We show that this increased efficacy is due to its higher surface density, driven by GSTs ability to oligomerize. We also show that these surfactant proteins slow droplet fusion and reduce average droplet size, as would be expected of a typical surfactant. Our findings quantitatively show how surfactant proteins can play a critical role in regulating the behavior of biomolecular condensates by modulating their interfacial tension. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=75 SRC="FIGDIR/small/660548v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@b0d206org.highwire.dtl.DTLVardef@acb567org.highwire.dtl.DTLVardef@b99e5dorg.highwire.dtl.DTLVardef@4bf12b_HPS_FORMAT_FIGEXP M_FIG C_FIG

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Favetta, B., Wang, H., Shi, Z., Schuster, B. S.. 2025-06-21. Amphiphilic protein surfactants reduce the interfacial tension of biomolecular condensates. https://doi.org/10.1101/2025.06.19.660548

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