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Single-molecule techniques to visualize and to characterize liquid-liquid phase separation and phase transition
Jinyao Ji1, Wenjuan Wang2, Chunlai Chen1
1School of Life Sciences, Beijing Advanced Innovation Center for Structural Biology, Beijing Frontier Research Center of Biological Structure, Tsinghua University, Beijing 100084, China.
Acta Biochimica Et Biophysica Sinica
|March 6, 2023
Summary
Single-molecule techniques offer powerful insights into liquid-liquid phase separation (LLPS) and the transition of biomolecular condensates from liquid-like to solid-like states, crucial for understanding cellular processes and diseases.
Area of Science:
- Biophysics
- Cell Biology
- Biochemistry
Background:
- Membraneless structures form via liquid-liquid phase separation (LLPS) in cells.
- Liquid-like condensates can transition to solid-like aggregations, linked to neurodegenerative diseases.
- Traditional ensemble methods distinguish condensates by morphology and dynamics, but lack molecular-level detail.
Purpose of the Study:
- To review single-molecule techniques for studying LLPS.
- To highlight their utility in manipulating LLPS and probing nanoscale mechanical properties.
- To demonstrate their power in monitoring molecular-level dynamic and thermodynamic properties.
Main Methods:
- Single-molecule force spectroscopy (e.g., AFM, optical tweezers).
- Single-molecule fluorescence spectroscopy (e.g., FRET, FCS).
- Single-particle tracking.
Main Results:
- Single-molecule techniques provide high sensitivity and mechanistic insights into LLPS.
- These methods enable manipulation of LLPS and characterization of nanoscale mechanical properties.
- Dynamic and thermodynamic properties at the molecular level can be precisely monitored.
Conclusions:
- Single-molecule techniques are essential tools for characterizing LLPS.
- They are uniquely suited to study the liquid-to-solid phase transition under physiological conditions.
- These methods advance our understanding of cellular organization and disease mechanisms.
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