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Crowder Chain Length Variability and Excluded Volume Effect on the Phase Separation Behavior of Mucin
Komal Kumari1, Anant Kumar Singh1, Priyankar Mandal1
1Department of Chemistry, Institute of Science, Banaras Hindu University, Varanasi, Uttar Pradesh 221005, India.
Insights
Mucin phase separation in cellular membranes is driven by excluded volume and crowder chain length. Shorter crowders increase mucin mobility within these condensed phases, impacting cancer progression.
Area of Science:
- Biochemistry
- Cell Biology
- Biophysics
Background:
- Phase separation is crucial for cellular functions and influenced by transmembrane proteins.
- Understanding transmembrane protein behavior in phase-separated states is vital.
Purpose of the Study:
- To investigate mucin behavior in cellular phase separation.
- To determine the roles of crowder chain length and excluded volume in this process.
Main Methods:
- Confocal microscopy to observe mucin partitioning.
- Fluorescence recovery after photobleaching to assess protein mobility.
- Theoretical modeling for excluded volume calculations.
Main Results:
- Mucin strongly partitions into the condensed phase, driven by hydrophobic and electrostatic interactions.
- Shorter crowder chains enhance mucin mobility in condensed phases.
- Excluded volume is critical for mucin phase separation under crowded conditions.
Conclusions:
- Mucin exhibits phase separation capabilities in crowded cellular environments.
- Excluded volume and crowder properties significantly modulate mucin's dynamic behavior.
- Findings enhance understanding of mucin's role in cancer progression.
Abstract:
Phase separation within cellular membranes, a critical process underpinning diverse cellular functions, is significantly influenced by transmembrane proteins. Therefore, elucidating the behavior of a transmembrane protein in its phase-separated state is of utmost importance. Our study explores mucin behavior in the cellular milieu, aiming to determine the role of crowder chain length and excluded volume in phase separation. Confocal microscopy images demonstrate the strong partitioning of mucin into the condensed phase influenced by hydrophobic and electrostatic interactions. Fluorescence recovery after photobleaching analysis revealed increased mobility in the presence of shorter chain length crowders, indicating the dynamic behavior of protein within condensed phases. Excluded volume calculation using the theoretical model emphasizes its importance in mucin phase separation under crowded conditions. Our findings underscore the ability of mucin to phase-separate under crowded conditions, highlighting the crucial role of excluded volume and enhancing our understanding of its involvement in cancer progression.
