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Ponceau S as a Targeted Modulator for Protein Liquid-Liquid Phase Separation.

Can Liu1, Qingpeng Zeng1, Xiuqin Ju1

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Ponceau S selectively induces and monitors protein liquid-liquid phase separation (LLPS) by binding hydrophobic regions. This controllable method offers new possibilities for biomolecular engineering and drug delivery applications.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Biophysics

Background:

  • Liquid-liquid phase separation (LLPS) is crucial for cellular organization and forming biomolecular condensates.
  • Existing methods for inducing protein LLPS often lack precise control over time and space.

Purpose of the Study:

  • To introduce Ponceau S as a novel, selective modulator for inducing and monitoring protein LLPS.
  • To investigate the mechanism by which Ponceau S influences protein phase separation.
  • To explore the tunability of Ponceau S-induced LLPS.

Main Methods:

  • Utilized Ponceau S to induce and observe LLPS in bovine serum albumin and lysozyme.
  • Analyzed Ponceau S fluorescence changes within protein-rich phases to infer molecular motion.
  • Adjusted salt concentrations and 1,6-hexanediol levels to modulate droplet dynamics.

Main Results:

  • Ponceau S effectively promoted protein complex formation into liquid droplets by interacting with hydrophobic regions.
  • Increased Ponceau S fluorescence within droplets indicated restricted molecular motion, confirming phase separation.
  • Phase separation dynamics, including droplet fusion and dissolution, were finely tunable via salt and 1,6-hexanediol concentrations.

Conclusions:

  • Ponceau S acts as a selective small-molecule modulator for protein LLPS.
  • Ponceau S enables precise regulation and monitoring of protein phase separation.
  • This approach offers potential for advancements in biomolecular engineering, drug delivery, and synthetic biology.