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Excipients Do Regulate Phase Separation in Lysozyme and Thus Also Its Hydration.
Partha Pyne1, Rajib Kumar Mitra1
1Department of Chemical, Biological and Macromolecular Sciences, S N Bose National Centre for Basic Sciences, Block JD, Sector III, Salt Lake, Kolkata 700106, India.
The Journal of Physical Chemistry Letters
|January 20, 2022
Summary
Protein hydration changes regulate liquid-liquid phase separation (LLPS). This study used THz-ATR-FTIR spectroscopy to analyze lysozyme hydration during LLPS, revealing how excipients like sucrose and BSA influence this crucial process.
Area of Science:
- Biophysics
- Protein Chemistry
- Spectroscopy
Background:
- Liquid-liquid phase separation (LLPS) is a critical process in cellular organization.
- The role of protein hydration changes in LLPS remains underexplored.
- Understanding LLPS regulation is key to comprehending cellular function and dysfunction.
Purpose of the Study:
- To experimentally investigate alterations in lysozyme hydration during LLPS.
- To explore the influence of various excipients on lysozyme LLPS and its hydration.
- To elucidate the role of hydrogen bonding in regulating protein LLPS.
Main Methods:
- Attenuated total reflection (ATR)-FTIR spectroscopy in the terahertz (THz) frequency region (1.5–21 THz).
- Experimental analysis of lysozyme hydration.
- Assessment of excipient effects (l-arginine, sucrose, BSA, ubiquitin) on LLPS and hydration.
Main Results:
- Significant changes in lysozyme hydration occur during LLPS.
- Sucrose was found to stabilize LLPS, while BSA inhibited it.
- Excipient effects varied, with l-arginine showing a subtle influence and ubiquitin's effect being concentration-dependent.
Conclusions:
- Alterations in protein hydration, specifically H-bond dynamics, are key regulators of LLPS.
- Excipients can modulate LLPS by affecting protein hydration profiles.
- THz-ATR-FTIR spectroscopy is a valuable tool for studying protein hydration in LLPS.
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