A Unified Description of Salt Effects on the Liquid–Liquid Phase Separation of Proteins

Duan, Chao; Wang, Rui

doi:10.1021/acscentsci.3c01372

Title: A Unified Description of Salt Effects on the Liquid–Liquid Phase Separation of Proteins

Journal Article · 07 February 2024 · ACS Central Science

DOI: https://doi.org/10.1021/acscentsci.3c01372 · OSTI ID:2370368

Duan, Chao ^[1];

^[2]

Univ. of California, Berkeley, CA (United States)
Univ. of California, Berkeley, CA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Protein aggregation via liquid-liquid phase separation (LLPS) is ubiquitous in nature and is intimately connected to many human diseases. Although it is widely known that the addition of salt has crucial impacts on the LLPS of proteins, full understanding of the salt effects remains an outstanding challenge. Here, we develop a molecular theory that systematically incorporates the self-consistent field theory for charged macromolecules into the solution thermodynamics. The electrostatic interaction, hydrophobicity, ion solvation, and translational entropy are included in a unified framework. Our theory fully captures the long-standing puzzles of the nonmonotonic salt concentration dependence and the specific ion effect. We find that proteins show salting-out at low salt concentrations due to ionic screening. The solubility follows the inverse Hofmeister series. In the high salt concentration regime, protein continues salting-out for small ions but turns to salting-in for larger ions, accompanied by the reversal of the Hofmeister series. We reveal that the solubility at high salt concentrations is determined by the competition between the solvation energy and translational entropy of the ion. Furthermore, we derive an analytical criterion for determining the boundary between the salting-in and salting-out regimes, which is in good agreement with experimental results for various proteins and salt ions.

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Research Organization:: Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences & Engineering Division (MSE)

Grant/Contract Number:: AC02-05CH11231

OSTI ID:: 2370368

Journal Information:: ACS Central Science, Journal Name: ACS Central Science Journal Issue: 2 Vol. 10; ISSN 2374-7943

Publisher:: American Chemical Society (ACS)Copyright Statement

Country of Publication:: United States

Language:: English

References (65)

Interactions of macromolecules with salt ions: An electrostatic theory for the Hofmeister effect Zhou, Huan‐Xiang Proteins: Structure, Function, and Bioinformatics, Vol. 61, Issue 1 https://doi.org/10.1002/prot.20500	journal	July 2005
How Hofmeister ion interactions affect protein stability Baldwin, R. L. Biophysical Journal, Vol. 71, Issue 4, p. 2056-2063 https://doi.org/10.1016/S0006-3495(96)79404-3	journal	October 1996
Liquid-Liquid Phase Separation of a Monoclonal Antibody and Nonmonotonic Influence of Hofmeister Anions Mason, Bruce D.; Zhang-van Enk, Jian; Zhang, Le Biophysical Journal, Vol. 99, Issue 11 https://doi.org/10.1016/j.bpj.2010.10.040	journal	December 2010
Considerations and Challenges in Studying Liquid-Liquid Phase Separation and Biomolecular Condensates Alberti, Simon; Gladfelter, Amy; Mittag, Tanja Cell, Vol. 176, Issue 3 https://doi.org/10.1016/j.cell.2018.12.035	journal	January 2019
Reversed Hofmeister series—The rule rather than the exception Schwierz, Nadine; Horinek, Dominik; Sivan, Uri Current Opinion in Colloid & Interface Science, Vol. 23 https://doi.org/10.1016/j.cocis.2016.04.003	journal	June 2016
Lower consolute boundaries of the nonionic surfactant C8E5 in aqueous alkali halide solutions: An approach to reproduce the effects of alkali halides on the cloud-point temperature Weckström, Kristian; Papageorgiou, Anastassios C. Journal of Colloid and Interface Science, Vol. 310, Issue 1 https://doi.org/10.1016/j.jcis.2007.01.045	journal	June 2007
Stimuli-responsive polymer as gate dielectric for organic transistor sensors Rullyani, Cut; Singh, Mriganka; Li, Sheng-Han Organic Electronics, Vol. 85 https://doi.org/10.1016/j.orgel.2020.105818	journal	October 2020
Liquid–liquid phase separation in cellular signaling systems Chong, P. Andrew; Forman-Kay, Julie D. Current Opinion in Structural Biology, Vol. 41 https://doi.org/10.1016/j.sbi.2016.08.001	journal	December 2016
Sequence Context and Complex Hofmeister Salt Interactions Dictate Phase Separation Propensity of Resilin-like Polypeptides Otis, James Brandt; Sharpe, Simon Biomacromolecules, Vol. 23, Issue 12 https://doi.org/10.1021/acs.biomac.2c01027	journal	November 2022
Electrostatic Interactions in Protein Structure, Folding, Binding, and Condensation Zhou, Huan-Xiang; Pang, Xiaodong Chemical Reviews, Vol. 118, Issue 4 https://doi.org/10.1021/acs.chemrev.7b00305	journal	January 2018
Beyond the Hofmeister Series: Ion-Specific Effects on Proteins and Their Biological Functions Okur, Halil I.; Hladílková, Jana; Rembert, Kelvin B. The Journal of Physical Chemistry B, Vol. 121, Issue 9 https://doi.org/10.1021/acs.jpcb.6b10797	journal	February 2017
50th Anniversary Perspective : A Perspective on Polyelectrolyte Solutions Muthukumar, M. Macromolecules, Vol. 50, Issue 24 https://doi.org/10.1021/acs.macromol.7b01929	journal	December 2017
Ureido-Derivatized Polymers Based on Both Poly(allylurea) and Poly( l -citrulline) Exhibit UCST-Type Phase Transition Behavior under Physiologically Relevant Conditions Shimada, Naohiko; Ino, Haruka; Maie, Kenji Biomacromolecules, Vol. 12, Issue 10 https://doi.org/10.1021/bm2010752	journal	October 2011
Specific Ion Effects at the Air/Water Interface Jungwirth, Pavel; Tobias, Douglas J. Chemical Reviews, Vol. 106, Issue 4 https://doi.org/10.1021/cr0403741	journal	April 2006
Hofmeister Phenomena: An Update on Ion Specificity in Biology Lo Nostro, Pierandrea; Ninham, Barry W. Chemical Reviews, Vol. 112, Issue 4 https://doi.org/10.1021/cr200271j	journal	January 2012
Classification of Intrinsically Disordered Regions and Proteins van der Lee, Robin; Buljan, Marija; Lang, Benjamin Chemical Reviews, Vol. 114, Issue 13 https://doi.org/10.1021/cr400525m	journal	December 2013
Dielectric properties of trypsin inhibitor and lysozyme calculated from molecular dynamics simulations Smith, Paul E.; Brunne, Roger M.; Mark, Alan E. The Journal of Physical Chemistry, Vol. 97, Issue 9 https://doi.org/10.1021/j100111a046	journal	March 1993
Dielectric Constants: Ethanol—Diethyl Ether and Urea—Water Solutions between 0 and 50° Wyman, Jeffries Journal of the American Chemical Society, Vol. 55, Issue 10 https://doi.org/10.1021/ja01337a029	journal	October 1933
Specific Ion Effects on the Water Solubility of Macromolecules: PNIPAM and the Hofmeister Series Zhang, Yanjie; Furyk, Steven; Bergbreiter, David E. Journal of the American Chemical Society, Vol. 127, Issue 41 https://doi.org/10.1021/ja0546424	journal	October 2005
Inverse Temperature Transition of a Biomimetic Elastin Model: Reactive Flux Analysis of Folding/Unfolding and Its Coupling to Solvent Dielectric Relaxation ^† Baer, Marcel; Schreiner, Eduard; Kohlmeyer, Axel The Journal of Physical Chemistry B, Vol. 110, Issue 8 https://doi.org/10.1021/jp054805a	journal	March 2006
Effects of Hofmeister Anions on the Phase Transition Temperature of Elastin-like Polypeptides Cho, Younhee; Zhang, Yanjie; Christensen, Trine The Journal of Physical Chemistry B, Vol. 112, Issue 44 https://doi.org/10.1021/jp8062977	journal	November 2008
Surface Tensions, Surface Potentials, and the Hofmeister Series of Electrolyte Solutions dos Santos, Alexandre P.; Diehl, Alexandre; Levin, Yan Langmuir, Vol. 26, Issue 13 https://doi.org/10.1021/la100604k	journal	July 2010
Possible Origin of the Inverse and Direct Hofmeister Series for Lysozyme at Low and High Salt Concentrations Boström, Mathias; Parsons, Drew F.; Salis, Andrea Langmuir, Vol. 27, Issue 15 https://doi.org/10.1021/la202023r	journal	August 2011
Reversed Anionic Hofmeister Series: The Interplay of Surface Charge and Surface Polarity Schwierz, Nadine; Horinek, Dominik; Netz, Roland R. Langmuir, Vol. 26, Issue 10 https://doi.org/10.1021/la904397v	journal	April 2010
Theory of Polymers in Poor Solvent: Phase Equilibrium and Nucleation Behavior Wang, Rui; Wang, Zhen-Gang Macromolecules, Vol. 45, Issue 15 https://doi.org/10.1021/ma301049m	journal	July 2012
Equilibrium cluster formation in concentrated protein solutions and colloids Stradner, Anna; Sedgwick, Helen; Cardinaux, Frédéric Nature, Vol. 432, Issue 7016 https://doi.org/10.1038/nature03109	journal	November 2004
Phase separation drives heterochromatin domain formation Strom, Amy R.; Emelyanov, Alexander V.; Mir, Mustafa Nature, Vol. 547, Issue 7662 https://doi.org/10.1038/nature22989	journal	June 2017
Phase behaviour of disordered proteins underlying low density and high permeability of liquid organelles Wei, Ming-Tzo; Elbaum-Garfinkle, Shana; Holehouse, Alex S. Nature Chemistry, Vol. 9, Issue 11 https://doi.org/10.1038/nchem.2803	journal	June 2017
Electrostatic control of block copolymer morphology Sing, Charles E.; Zwanikken, Jos W.; Olvera de la Cruz, Monica Nature Materials, Vol. 13, Issue 7 https://doi.org/10.1038/nmat4001	journal	June 2014
Polymer physics of intracellular phase transitions Brangwynne, Clifford P.; Tompa, Peter; Pappu, Rohit V. Nature Physics, Vol. 11, Issue 11 https://doi.org/10.1038/nphys3532	journal	November 2015
The amyloid state and its association with protein misfolding diseases Knowles, Tuomas P. J.; Vendruscolo, Michele; Dobson, Christopher M. Nature Reviews Molecular Cell Biology, Vol. 15, Issue 6 https://doi.org/10.1038/nrm3810	journal	May 2014
Signal integration through blending, bolstering and bifurcating of intracellular information Pawson, Catherine T.; Scott, John D. Nature Structural & Molecular Biology, Vol. 17, Issue 6 https://doi.org/10.1038/nsmb.1843	journal	May 2010
Reentrant liquid condensate phase of proteins is stabilized by hydrophobic and non-ionic interactions Krainer, Georg; Welsh, Timothy J.; Joseph, Jerelle A. Nature Communications, Vol. 12, Issue 1 https://doi.org/10.1038/s41467-021-21181-9	journal	February 2021
Generic nature of the condensed states of proteins Fuxreiter, Monika; Vendruscolo, Michele Nature Cell Biology, Vol. 23, Issue 6 https://doi.org/10.1038/s41556-021-00697-8	journal	June 2021
α-Synuclein aggregation nucleates through liquid–liquid phase separation Ray, Soumik; Singh, Nitu; Kumar, Rakesh Nature Chemistry, Vol. 12, Issue 8 https://doi.org/10.1038/s41557-020-0465-9	journal	June 2020
Weakly hydrated anions bind to polymers but not monomers in aqueous solutions Rogers, Bradley A.; Okur, Halil I.; Yan, Chuanyu Nature Chemistry, Vol. 14, Issue 1 https://doi.org/10.1038/s41557-021-00805-z	journal	November 2021
A framework for understanding the functions of biomolecular condensates across scales Lyon, Andrew S.; Peeples, William B.; Rosen, Michael K. Nature Reviews Molecular Cell Biology, Vol. 22, Issue 3 https://doi.org/10.1038/s41580-020-00303-z	journal	November 2020
Biomolecular condensates at the nexus of cellular stress, protein aggregation disease and ageing Alberti, Simon; Hyman, Anthony A. Nature Reviews Molecular Cell Biology, Vol. 22, Issue 3 https://doi.org/10.1038/s41580-020-00326-6	journal	January 2021
The nucleolus as a multiphase liquid condensate Lafontaine, Denis L. J.; Riback, Joshua A.; Bascetin, Rümeyza Nature Reviews Molecular Cell Biology, Vol. 22, Issue 3 https://doi.org/10.1038/s41580-020-0272-6	journal	September 2020
Composition-dependent thermodynamics of intracellular phase separation Riback, Joshua A.; Zhu, Lian; Ferrolino, Mylene C. Nature, Vol. 581, Issue 7807 https://doi.org/10.1038/s41586-020-2256-2	journal	May 2020
Nucleation landscape of biomolecular condensates Shimobayashi, Shunsuke F.; Ronceray, Pierre; Sanders, David W. Nature, Vol. 599, Issue 7885 https://doi.org/10.1038/s41586-021-03905-5	journal	September 2021
Capillary forces generated by biomolecular condensates Gouveia, Bernardo; Kim, Yoonji; Shaevitz, Joshua W. Nature, Vol. 609, Issue 7926 https://doi.org/10.1038/s41586-022-05138-6	journal	September 2022
Models and mechanisms of Hofmeister effects in electrolyte solutions, and colloid and protein systems revisited Salis, Andrea; Ninham, Barry W. Chem. Soc. Rev., Vol. 43, Issue 21 https://doi.org/10.1039/C4CS00144C	journal	January 2014
Tuning the collapse transition of weakly charged polymers by ion-specific screening and adsorption Chudoba, Richard; Heyda, Jan; Dzubiella, Joachim Soft Matter, Vol. 14, Issue 47 https://doi.org/10.1039/C8SM01646A	journal	January 2018
Effects of ion solvation on phase equilibrium and interfacial tension of liquid mixtures Wang, Rui; Wang, Zhen-Gang The Journal of Chemical Physics, Vol. 135, Issue 1 https://doi.org/10.1063/1.3607969	journal	July 2011
Conformation of a single polyelectrolyte in poor solvents Duan, Chao; Li, Weihua; Wang, Rui The Journal of Chemical Physics, Vol. 153, Issue 6 https://doi.org/10.1063/5.0017371	journal	August 2020
The inverse and direct Hofmeister series for lysozyme Zhang, Y.; Cremer, P. S. Proceedings of the National Academy of Sciences, Vol. 106, Issue 36 https://doi.org/10.1073/pnas.0907616106	journal	August 2009
Protein aggregation in salt solutions Kastelic, Miha; Kalyuzhnyi, Yurij V.; Hribar-Lee, Barbara Proceedings of the National Academy of Sciences, Vol. 112, Issue 21 https://doi.org/10.1073/pnas.1507303112	journal	May 2015
Equilibrium constants and free energies in unfolding of proteins in urea solutions Klotz, I. M. Proceedings of the National Academy of Sciences, Vol. 93, Issue 25 https://doi.org/10.1073/pnas.93.25.14411	journal	December 1996
The self-assembly, aggregation and phase transitions of food protein systems in one, two and three dimensions Mezzenga, Raffaele; Fischer, Peter Reports on Progress in Physics, Vol. 76, Issue 4 https://doi.org/10.1088/0034-4885/76/4/046601	journal	March 2013
Beyond standard Poisson–Boltzmann theory: ion-specific interactions in aqueous solutions Ben-Yaakov, Dan; Andelman, David; Harries, Daniel Journal of Physics: Condensed Matter, Vol. 21, Issue 42 https://doi.org/10.1088/0953-8984/21/42/424106	journal	September 2009
Soft Matter Physics Doi, Masao Oxford University Press https://doi.org/10.1093/acprof:oso/9780199652952.001.0001	book	January 2013
Fluctuation in electrolyte solutions: The self energy Wang, Zhen-Gang Physical Review E, Vol. 81, Issue 2 https://doi.org/10.1103/PhysRevE.81.021501	journal	February 2010
Continuous Self-Energy of Ions at the Dielectric Interface Wang, Rui; Wang, Zhen-Gang Physical Review Letters, Vol. 112, Issue 13 https://doi.org/10.1103/PhysRevLett.112.136101	journal	April 2014
Self-Consistent Description of Vapor-Liquid Interface in Ionic Fluids Agrawal, Nikhil R.; Wang, Rui Physical Review Letters, Vol. 129, Issue 22 https://doi.org/10.1103/PhysRevLett.129.228001	journal	November 2022
Electrostatics-Induced Nucleated Conformational Transition of Protein Aggregation Duan, Chao; Wang, Rui Physical Review Letters, Vol. 130, Issue 15 https://doi.org/10.1103/PhysRevLett.130.158401	journal	April 2023
Like dissolves like: A first-principles theory for predicting liquid miscibility and mixture dielectric constant Zhuang, Bilin; Ramanauskaite, Gabriele; Koa, Zhao Yuan Science Advances, Vol. 7, Issue 7 https://doi.org/10.1126/sciadv.abe7275	journal	February 2021
Germline P Granules Are Liquid Droplets That Localize by Controlled Dissolution/Condensation Brangwynne, C. P.; Eckmann, C. R.; Courson, D. S. Science, Vol. 324, Issue 5935 https://doi.org/10.1126/science.1172046	journal	May 2009
Cytoplasmic protein aggregates interfere with nucleocytoplasmic transport of protein and RNA Woerner, A. C.; Frottin, F.; Hornburg, D. Science, Vol. 351, Issue 6269 https://doi.org/10.1126/science.aad2033	journal	December 2015
Liquid phase condensation in cell physiology and disease Shin, Yongdae; Brangwynne, Clifford P. Science, Vol. 357, Issue 6357 https://doi.org/10.1126/science.aaf4382	journal	September 2017
ATP as a biological hydrotrope Patel, Avinash; Malinovska, Liliana; Saha, Shambaditya Science, Vol. 356, Issue 6339 https://doi.org/10.1126/science.aaf6846	journal	May 2017
Valence and patterning of aromatic residues determine the phase behavior of prion-like domains Martin, Erik W.; Holehouse, Alex S.; Peran, Ivan Science, Vol. 367, Issue 6478 https://doi.org/10.1126/science.aaw8653	journal	February 2020
Protein Misfolding, Amyloid Formation, and Human Disease: A Summary of Progress Over the Last Decade Chiti, Fabrizio; Dobson, Christopher M. Annual Review of Biochemistry, Vol. 86, Issue 1 https://doi.org/10.1146/annurev-biochem-061516-045115	journal	June 2017
Physical Principles Underlying the Complex Biology of Intracellular Phase Transitions Choi, Jeong-Mo; Holehouse, Alex S.; Pappu, Rohit V. Annual Review of Biophysics, Vol. 49, Issue 1 https://doi.org/10.1146/annurev-biophys-121219-081629	journal	May 2020
Chemistry of Hofmeister Anions and Osmolytes Zhang, Yanjie; Cremer, Paul S. Annual Review of Physical Chemistry, Vol. 61, Issue 1 https://doi.org/10.1146/annurev.physchem.59.032607.093635	journal	March 2010

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