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Coacervate droplets as pH-regionalized protocells.

Chong Wang1,2, Zhonglin Fang2, Linyi Zhang2

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Liquid-liquid phase separation (LLPS) in peptide coacervates creates distinct pH microenvironments within membrane-less organelles (MLOs). This pH modulation is crucial for MLO function and can be harnessed for protocell applications.

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Area of Science:

  • Biochemistry
  • Cell Biology
  • Biophysics

Background:

  • pH regulation in membrane-less organelles (MLOs) is poorly understood.
  • Mechanisms differ significantly from membrane-bound organelles.
  • Directly controlling MLO interfaces is challenging.

Purpose of the Study:

  • Investigate the role of liquid-liquid phase separation (LLPS) in MLO pH regulation.
  • Develop an in vitro model for studying MLO pH dynamics.
  • Explore potential applications of pH-modulated coacervates.

Main Methods:

  • Constructed peptide-based coacervate droplets using microfluidics.
  • Analyzed H+/OH- distributions within coacervates.
  • Observed pH changes upon LLPS disruption.
  • Incorporated enzymes to tune local pH.

Main Results:

  • Charged polymers in coacervates create pH-regionalized microenvironments.
  • LLPS is essential for maintaining pH differences, mimicking nucleolar conditions.
  • pH differences diminish upon LLPS destruction.
  • Enzymes enabled fine-tuning of local pH within coacervates.

Conclusions:

  • LLPS plays a critical role in pH modulation within MLOs.
  • Coacervate droplets serve as a viable in vitro model for MLO pH studies.
  • Coacervates show potential as protocells for molecular biology applications like PCR and IVTT.