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Three-Phase Coexistence in Binary Charged Lipid Membranes in a Hypotonic Solution.

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We found three coexisting phases in charged lipid membranes (dioleoylphosphatidylserine and dipalmitoylphosphatidylcholine) under osmotic stress. This unexpected three-phase coexistence was confirmed by microscopy and molecular dynamics simulations.

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Area of Science:

  • Membrane biophysics
  • Lipid phase behavior
  • Soft matter physics

Background:

  • Charged lipid membranes typically resist phase separation due to electrostatic repulsion.
  • Osmotic stress can influence lipid membrane organization and domain formation.
  • Understanding lipid mixtures is crucial for cell membrane function and biomaterial design.

Purpose of the Study:

  • To investigate the phase separation behavior of dioleoylphosphatidylserine (DOPS) and dipalmitoylphosphatidylcholine (DPPC) mixtures.
  • To explore the effect of osmotic stress on charged lipid membrane phase separation.
  • To elucidate the mechanisms stabilizing complex phase behavior in binary lipid systems.

Main Methods:

  • Giant unilamellar vesicles (GUVs) were prepared with DOPS/DPPC mixtures.
  • Fluorescence and confocal laser scanning microscopy were employed to visualize phase separation.
  • Coarse-grained molecular dynamics simulations were performed to confirm experimental findings.

Main Results:

  • A three-phase coexistence was observed in DOPS/DPPC binary mixtures under hypotonic conditions.
  • The identified phases were DPPC-rich, dissociated DOPS-rich, and nondissociated DOPS-rich.
  • The coexistence of differently ionized DOPS domains was experimentally verified by particle adsorption and computationally confirmed.

Conclusions:

  • Osmotic stress can induce complex phase separation in charged lipid membranes, leading to a quasi-ternary system.
  • Hydrogen bonding between protonated DOPS and reduced electrostatic interactions stabilize the observed three-phase coexistence.
  • This study reveals novel phase behavior in charged lipid mixtures, relevant to biological membranes and nanotechnology.