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A light-switching pyrene probe to detect phase-separated biomolecules
Masaharu Hazawa1,2,3, Shogo Amemori1,2,4, Yoshio Nishiyama2
1Institute for Frontier Science Initiative, Kanazawa University, Kanazawa, Ishikawa, Japan.
Iscience
|August 13, 2021
Summary
A new probe, Pyr-A, allows scientists to study liquid-liquid phase separation (LLPS) by measuring changes in viscosity and hydrophobicity. This tool helps understand how LLPS regulates biological processes in protein droplets and centrosomes.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Biomolecular liquid-liquid phase separation (LLPS) is crucial for regulating biological processes.
- Limited tools exist to probe the physicochemical properties of phase-separated condensates.
- Understanding LLPS is key to deciphering cellular organization and function.
Purpose of the Study:
- To introduce a novel light-switching dipyrene probe (Pyr-A) for studying LLPS.
- To characterize the biophysical properties of phase-separated condensates.
- To enhance the understanding of LLPS in biological systems.
Main Methods:
- Development of a light-switching dipyrene probe (Pyr-A).
- Utilizing fluorescent microscopic imaging and ratiometric measurements.
- Studying *in vitro* protein droplets and *in vivo* centrosomes.
Main Results:
- Pyr-A exhibits distinct fluorescence spectra in hydrophobic/viscous versus aqueous environments.
- Increased size of protein droplets correlated with increased hydrophobicity and viscosity.
- Centrosome maturation involved increased hydrophobicity and viscosity.
Conclusions:
- Pyr-A is an effective tool for characterizing LLPS.
- The probe provides insights into the biophysical changes during LLPS.
- This work advances the understanding of phase separation in biological functions.
