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Micro-Raman spectroscopic analysis of liquid-liquid phase separation

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Title
Micro-Raman spectroscopic analysis of liquid-liquid phase separation
Author(s)
Suin Choi; So Yeon Chun; Kyungwon Kwak; Minhaeng Cho
Publication Date
2023-03
Journal
PHYSICAL CHEMISTRY CHEMICAL PHYSICS, v.25, no.13, pp.9051 - 9060
Publisher
ROYAL SOC CHEMISTRY
Abstract
Liquid-liquid phase separation (LLPS) plays a significant role in various biological processes, including the formation of membraneless organelles and pathological protein aggregation. Although many studies have found various factors that modulate the LLPS process or the liquid-to-solid phase transition (LSPT) using microscopy or fluorescence-based methods, the molecular mechanistic details underlying LLPS and protein aggregation within liquid droplets remain uncharacterized. Therefore, structural information on proteins inside liquid droplets is required to understand the mechanistic link to amyloid formation. In the present study, we monitored droplet formation related to protein fibrillation using micro-Raman spectroscopy in combination with differential interference contrast (DIC) microscopy to study the conformational change in proteins and the hydrogen-bonding (H-bonding) structure of water during LLPS. Interestingly, we found that the O-D stretching band for water (HOD in H2O) inside the droplets exhibited a distinct Raman spectrum from that of the bulk water, suggesting that the time-dependent change in the hydration environment in the protein droplets during the process of LLPS can be studied. These results demonstrate that the superior spatial resolution of micro-Raman spectroscopy offers significant advantages in investigating the molecular mechanisms of LLPS and following LSPT processes.
URI
https://pr.ibs.re.kr/handle/8788114/13738
DOI
10.1039/d2cp05115j
ISSN
1463-9076
Appears in Collections:
Center for Molecular Spectroscopy and Dynamics(분자 분광학 및 동력학 연구단) > 1. Journal Papers (저널논문)
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