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Related Experiment Video

Updated: Oct 17, 2025

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Assembly of model postsynaptic densities involves interactions auxiliary to stoichiometric binding.

Yi-Hsuan Lin1, Haowei Wu2, Bowen Jia2

  • 1Department of Biochemistry, University of Toronto, Toronto, Ontario, Canada; Molecular Medicine, The Hospital for Sick Children, Toronto, Ontario, Canada.

Biophysical Journal
|October 12, 2021
PubMed
Summary

Biomolecular condensates form through liquid-liquid phase separation (LLPS) driven by protein interactions. This study shows that interactions beyond simple stoichiometric complexes, like weaker or higher-order ones, are crucial for SynGAP/PSD-95 condensate assembly.

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Neuroscience

Background:

  • Functional biomolecular condensates assemble via liquid-liquid phase separation (LLPS) of proteins with modular domains.
  • Understanding LLPS-driving domain-domain interactions is key, questioning if dilute solution interactions predict condensed phase behavior.

Purpose of the Study:

  • To investigate whether interactions driving LLPS are exclusively those forming discrete complexes in dilute solutions.
  • To analyze the role of various interaction types in the assembly of SynGAP/PSD-95 condensates, a model for neuronal postsynaptic densities (PSDs).

Main Methods:

  • Development of a mean-field LLPS theory for two stoichiometrically constrained solute species.
  • Application of the theory to neuronal proteins SynGAP and PSD-95, comparing predictions with experimental data.
  • Testing theoretical models with varying interaction scenarios, including stoichiometric and auxiliary interactions.

Main Results:

  • A theory imposing a fixed 3:2 SynGAP/PSD-95 stoichiometry failed to match experimental phase diagrams.
  • Models incorporating auxiliary SynGAP-PSD-95, SynGAP-SynGAP, and PSD-95-PSD-95 interactions aligned with experimental tie-line patterns.
  • Evidence suggests undocumented weaker or higher-order interactions contribute to SynGAP/PSD-95 condensate formation.

Conclusions:

  • LLPS-driven condensate assembly likely involves a synergy between specific stoichiometric binding and stochastic, multivalent interactions.
  • Future research should focus on identifying the auxiliary interactions governing PSD-like LLPS.
  • The findings challenge the assumption that dilute solution complex formation solely dictates LLPS behavior.