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Updated: May 6, 2026
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Author Spotlight: Evaluation of Protein-Condensate Dynamics in Live Human Cells
Published on: January 5, 2024
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Using Solution NMR to Characterize Biomolecular Condensates Under Biphasic Conditions
Mihajlo Novakovic1, Johannes Schmoll1, Leonidas Emmanouilidis1
1Department of Biology, Institute of Biochemistry, ETH Zurich.
Journal of Visualized Experiments : Jove
|May 4, 2026
Summary
Nuclear magnetic resonance (NMR) spectroscopy offers label-free methods to study biomolecular condensates. New NMR techniques, REDIFINE and CONDENSE-MT, characterize condensate dynamics and composition without tags.
Area of Science:
- Biophysics
- Molecular Biology
- Biochemistry
Background:
- Biomolecular condensates, formed via liquid-liquid phase separation (LLPS), are crucial for cellular organization and biochemical regulation.
- Investigating condensate composition, dynamics, and internal structure without external tags presents significant challenges.
Purpose of the Study:
- To develop and validate novel Nuclear Magnetic Resonance (NMR) methodologies for label-free characterization of biomolecular condensates.
- To provide quantitative insights into the physicochemical properties and dynamics of condensates in their native biphasic state.
Main Methods:
- Utilized two complementary NMR approaches: REstricted DIffusion of INvisible speciEs (REDIFINE) for dynamic condensates and CONdensate DEtectioN by SEmi-solid Magnetization Transfer (CONDENSE-MT) for rigid condensates.
- REDIFINE quantifies phase partitioning, droplet size, interface permeability, and molecular exchange rates using diffusion-exchange contrast.
- CONDENSE-MT employs water-detected magnetization transfer to assess partitioning, molecular tumbling, hydration, and bound water in NMR-invisible condensates.
Main Results:
- Demonstrated the capability of REDIFINE to measure exchange rates and droplet properties for dynamic condensates.
- Showcased CONDENSE-MT's effectiveness in characterizing the biophysical properties of rigid condensates.
- Integrated both methods to provide a comprehensive, multidimensional view of condensate structure and dynamics under near-native conditions.
Conclusions:
- These advanced NMR techniques offer powerful, label-free tools for studying biomolecular phase separation.
- The developed methodologies enable a deeper understanding of condensate physicochemical properties and their link to biological functions and diseases.
- Expanded the NMR toolbox for investigating complex cellular organization driven by phase separation.
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