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Competitive protein recruitment in artificial cells.

Thijs W van Veldhuisen1, Madelief A M Verwiel1, Sebastian Novosedlik1

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Artificial cells engineered with varying protein-binding affinities enable directed communication. This system facilitates controlled protein exchange between distinct cell populations, mimicking intercellular signaling.

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Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Cellular Communication

Background:

  • Living cells dynamically adjust responses to environmental signals via sensitivity modulation.
  • Artificial cells offer models for studying intercellular communication, but differentiated populations are less explored.

Purpose of the Study:

  • To demonstrate affinity-regulated protein exchange in coacervate-based artificial cells.
  • To engineer a communication system for directed protein transport between artificial cell populations.

Main Methods:

  • Utilized coacervate-based artificial cells loaded with different 14-3-3 protein isoforms with varying protein-protein interaction (PPI) affinities.
  • Engineered client protein affinity switching via phosphorylation to control binding and release.
  • Established competitive binding dynamics for directed molecular exchange.

Main Results:

  • Directed client peptide uptake by coacervates with higher 14-3-3 binding affinity was achieved.
  • Phosphorylation-induced affinity changes enabled the release of weaker binding partners.
  • A communication system was constructed, enabling protein transport from strongly to weakly recruiting artificial cells.

Conclusions:

  • Affinity engineering and competitive binding are effective strategies for directed protein uptake and exchange in artificial cells.
  • Demonstrated a novel communication system in artificial cells using protein-binding affinity modulation.
  • Highlights the potential of engineered artificial cells for complex biological process modeling.