Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation

Biomolecular condensation is an important mechanism of cellular compartmentalization without membranes. Formation of liquid-like condensates of biomolecules involves protein-protein interactions working in tandem with protein-water interactions. The balance of these interactions in condensate-formin...

Full description

Saved in:
Bibliographic Details
Main Authors: Artur Czajkowski, Abhirami Udayabanu, Manthan Raj, Likhitha Ch. P. Pulibandla, Marija Tursunović, Marcus Jahnel, Ellen M. Adams
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-03-01
Series:Frontiers in Nanotechnology
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fnano.2025.1556384/full
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850232651196137472
author Artur Czajkowski
Artur Czajkowski
Abhirami Udayabanu
Abhirami Udayabanu
Manthan Raj
Manthan Raj
Likhitha Ch. P. Pulibandla
Likhitha Ch. P. Pulibandla
Marija Tursunović
Marcus Jahnel
Marcus Jahnel
Ellen M. Adams
Ellen M. Adams
author_facet Artur Czajkowski
Artur Czajkowski
Abhirami Udayabanu
Abhirami Udayabanu
Manthan Raj
Manthan Raj
Likhitha Ch. P. Pulibandla
Likhitha Ch. P. Pulibandla
Marija Tursunović
Marcus Jahnel
Marcus Jahnel
Ellen M. Adams
Ellen M. Adams
author_sort Artur Czajkowski
collection DOAJ
description Biomolecular condensation is an important mechanism of cellular compartmentalization without membranes. Formation of liquid-like condensates of biomolecules involves protein-protein interactions working in tandem with protein-water interactions. The balance of these interactions in condensate-forming proteins is impacted by multiple factors inside of a living organism. This work investigates the effects of post-translational modifications (PTMs) and salt concentration as two such perturbing factors on the protein Fused in Sarcoma (FUS), an RNA binding protein. The protein was obtained from two expression systems differing by their capability to add PTMs to the protein, bacterial and insect cell. Attenuated total reflection Terahertz spectroscopy is used to probe the solvation behavior in condensates formed from FUS protein with and without PTMs at 100 mM and 2.5 M KCl. The results show that while PTMs impact the phase-separating propensity, they do not alter protein solvation in the condensate. On the other hand, salt concentration was found to alter the stiffness of the water hydrogen bond network. These findings have implications for biomolecular condensates chemistry, showing that condensate molecular organization is perturbed by fluctuations in solvent properties.
format Article
id doaj-art-711167b4bdef48fa8cf0d400fca9c9a2
institution OA Journals
issn 2673-3013
language English
publishDate 2025-03-01
publisher Frontiers Media S.A.
record_format Article
series Frontiers in Nanotechnology
spelling doaj-art-711167b4bdef48fa8cf0d400fca9c9a22025-08-20T02:03:07ZengFrontiers Media S.A.Frontiers in Nanotechnology2673-30132025-03-01710.3389/fnano.2025.15563841556384Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensationArtur Czajkowski0Artur Czajkowski1Abhirami Udayabanu2Abhirami Udayabanu3Manthan Raj4Manthan Raj5Likhitha Ch. P. Pulibandla6Likhitha Ch. P. Pulibandla7Marija Tursunović8Marcus Jahnel9Marcus Jahnel10Ellen M. Adams11Ellen M. Adams12Cluster of Excellence Physics of Life, TU Dresden, Dresden, GermanyInstitute of Resource Ecology, Helmholtz Zentrum Dresden Rossendorf, Dresden, GermanyCluster of Excellence Physics of Life, TU Dresden, Dresden, GermanyInstitute of Resource Ecology, Helmholtz Zentrum Dresden Rossendorf, Dresden, GermanyCluster of Excellence Physics of Life, TU Dresden, Dresden, GermanyInstitute of Resource Ecology, Helmholtz Zentrum Dresden Rossendorf, Dresden, GermanyCluster of Excellence Physics of Life, TU Dresden, Dresden, GermanyInstitute of Resource Ecology, Helmholtz Zentrum Dresden Rossendorf, Dresden, GermanyFaculty of Chemistry, University of Belgrade, Belgrade, SerbiaCluster of Excellence Physics of Life, TU Dresden, Dresden, GermanyBiotechnology Center (BIOTEC), Center for Molecular and Cellular Bioengineering (CMCB), TU Dresden, Dresden, GermanyCluster of Excellence Physics of Life, TU Dresden, Dresden, GermanyInstitute of Resource Ecology, Helmholtz Zentrum Dresden Rossendorf, Dresden, GermanyBiomolecular condensation is an important mechanism of cellular compartmentalization without membranes. Formation of liquid-like condensates of biomolecules involves protein-protein interactions working in tandem with protein-water interactions. The balance of these interactions in condensate-forming proteins is impacted by multiple factors inside of a living organism. This work investigates the effects of post-translational modifications (PTMs) and salt concentration as two such perturbing factors on the protein Fused in Sarcoma (FUS), an RNA binding protein. The protein was obtained from two expression systems differing by their capability to add PTMs to the protein, bacterial and insect cell. Attenuated total reflection Terahertz spectroscopy is used to probe the solvation behavior in condensates formed from FUS protein with and without PTMs at 100 mM and 2.5 M KCl. The results show that while PTMs impact the phase-separating propensity, they do not alter protein solvation in the condensate. On the other hand, salt concentration was found to alter the stiffness of the water hydrogen bond network. These findings have implications for biomolecular condensates chemistry, showing that condensate molecular organization is perturbed by fluctuations in solvent properties.https://www.frontiersin.org/articles/10.3389/fnano.2025.1556384/fullbiomolecular condensatesprotein solvationTHz spectroscopyreentrant phase separationpost-translational modifications
spellingShingle Artur Czajkowski
Artur Czajkowski
Abhirami Udayabanu
Abhirami Udayabanu
Manthan Raj
Manthan Raj
Likhitha Ch. P. Pulibandla
Likhitha Ch. P. Pulibandla
Marija Tursunović
Marcus Jahnel
Marcus Jahnel
Ellen M. Adams
Ellen M. Adams
Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation
Frontiers in Nanotechnology
biomolecular condensates
protein solvation
THz spectroscopy
reentrant phase separation
post-translational modifications
title Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation
title_full Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation
title_fullStr Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation
title_full_unstemmed Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation
title_short Protein modifications and ionic strength show the difference between protein-mediated and solvent-mediated regulation of biomolecular condensation
title_sort protein modifications and ionic strength show the difference between protein mediated and solvent mediated regulation of biomolecular condensation
topic biomolecular condensates
protein solvation
THz spectroscopy
reentrant phase separation
post-translational modifications
url https://www.frontiersin.org/articles/10.3389/fnano.2025.1556384/full
work_keys_str_mv AT arturczajkowski proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT arturczajkowski proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT abhiramiudayabanu proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT abhiramiudayabanu proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT manthanraj proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT manthanraj proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT likhithachppulibandla proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT likhithachppulibandla proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT marijatursunovic proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT marcusjahnel proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT marcusjahnel proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT ellenmadams proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation
AT ellenmadams proteinmodificationsandionicstrengthshowthedifferencebetweenproteinmediatedandsolventmediatedregulationofbiomolecularcondensation