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Area of Science:

  • Physical Chemistry
  • Biophysics
  • Spectroscopy

Background:

  • Liquid-liquid phase separation (LLPS) is a crucial process in biological organization, involving the reversible compartmentalization of protein solutions.
  • Understanding the thermodynamic driving forces behind LLPS is essential for manipulating this process for biological and medical applications.
  • Water's role in LLPS has been traditionally viewed as passive, but its active participation is increasingly recognized.

Purpose of the Study:

  • To develop a general thermodynamic model quantifying the driving forces of LLPS based on water's behavior.
  • To elucidate the distinct roles of different water populations (cavity-wrap and bound water) in driving LLPS.
  • To introduce a spectroscopic method for mapping and tuning LLPS phenomena.

Main Methods:

  • Utilized terahertz (THz) spectroscopy to measure characteristic spectral features associated with water hydration.
  • Analyzed the amplitudes of THz features corresponding to two distinct water populations: 'cavity-wrap' and 'bound' water.
  • Developed a THz-phase diagram to correlate spectroscopic changes with thermodynamic parameters (temperature, concentration).

Main Results:

  • Demonstrated that LLPS is thermodynamically driven by changes in hydration entropy and enthalpy.
  • Showed that 'cavity-wrap' water released during LLPS increases entropy, while 'bound' water retained during LLPS is enthalpically favorable.
  • Successfully mapped spectroscopic and thermodynamic changes using a novel THz-phase diagram.

Conclusions:

  • LLPS is actively driven by the distinct hydration behaviors of hydrophobic and hydrophilic patches.
  • The developed THz-spectroscopic model provides a precise understanding of these hydration driving forces.
  • This approach offers a rational strategy for tuning LLPS by adjusting experimental parameters.