Effect of molecular weight distribution on the liquid–liquid phase separation behavior of polydispersed polyethylene solutions at high temperatures

2011 ◽  
Vol 305 (2) ◽  
pp. 152-160 ◽  
Author(s):  
Masashi Haruki ◽  
Kunio Nakanishi ◽  
Shigemitsu Mano ◽  
Shin-ichi Kihara ◽  
Shigeki Takishima
Keyword(s):  
Molecular Weight ◽  
Phase Separation ◽  
Liquid Phase ◽  
Molecular Weight Distribution ◽  
High Temperatures ◽  
Weight Distribution ◽  
Liquid Phase Separation ◽  
Phase Separation Behavior ◽  
Separation Behavior ◽  
Liquid Liquid Phase Separation

scholarly journals Liquid-liquid phase separation and liquid-to-solid transition mediate α-synuclein amyloid fibril containing hydrogel formation

2019 ◽  
Author(s):  
Soumik Ray ◽  
Nitu Singh ◽  
Satyaprakash Pandey ◽  
Rakesh Kumar ◽  
Laxmikant Gadhe ◽  
...  
Keyword(s):  
Phase Separation ◽  
Liquid Phase ◽  
Liquid Droplet ◽  
Liquid Phase Separation ◽  
Amyloid Formation ◽  
Liquid To Solid Transition ◽  
Phase Separation Behavior ◽  
Separation Behavior ◽  
Liquid Liquid Phase Separation

SUMMARYα-Synuclein (α-Syn) aggregation and amyloid formation is directly linked with Parkinson’s disease (PD) pathogenesis. However, the early events involved in this process remain unclear. Here, using in vitro reconstitution and cellular model, we show that liquid-liquid phase separation (LLPS) of α-Syn precedes its aggregation. In particular, in vitro generated α-Syn liquid-like droplets eventually undergo a liquid-to-solid transition and form amyloid-hydrogel containing oligomers and fibrillar species. Factors known to aggravate α-Syn aggregation such as low pH, phosphomimic substitution, and familial PD mutation also promote α-Syn LLPS and its subsequent maturation. We further demonstrate α-Syn liquid droplet formation in cells, under oxidative stress. These cellular α-Syn droplets eventually transform into perinuclear aggresomes, the process regulated by microtubules. The present work provides detailed insights into the phase separation behavior of natively unstructured α-Syn and its conversion to a disease-associated aggregated state, which is highly relevant in PD pathogenesis.

scholarly journals A generic approach for studying the kinetics of liquid-liquid phase separation under near-native conditions

2019 ◽  
Author(s):  
Joris van Lindt ◽  
Anna Bratek-Skicki ◽  
Donya Pakravan ◽  
Ludo Van Den Bosch ◽  
Dominique Maes ◽  
...  
Keyword(s):  
Phase Separation ◽  
Liquid Phase ◽  
Cell Biology ◽  
Low Complexity ◽  
Liquid Phase Separation ◽  
Ph Values ◽  
Phase Separation Behavior ◽  
Kinetics Of ◽  
Separation Behavior ◽  
Liquid Liquid Phase Separation

Understanding the kinetics and underlying physicochemical forces of liquid-liquid phase separation (LLPS) is of paramount importance in cell biology, requiring reproducible methods for the analysis of often severely aggregation-prone proteins. Frequently applied approaches, such as dilution of the protein from an urea-containing solution or cleavage of its fused solubility tag, however, often lead to very different kinetic behaviors. Here we suggest that at extreme pH values even proteins such as the low-complexity domain (LCD) of hnRNPA2, TDP-43, and NUP-98 can be kept in solution, and then their LLPS can be induced by a jump to native pH, resulting in a system that can be easily controlled. This approach represents a generic method for studying LLPS under near native conditions, providing a platform for studying the phase-separation behavior of diverse proteins.

Stereo-chemical contributions to the glass transition and liquid–liquid phase separation in high molecular weight poly(N-vinyl carbazole)

RSC Advances ◽  
2016 ◽  
Vol 6 (35) ◽  
pp. 29326-29333 ◽  
Author(s):  
Abdul G. Al Lafi ◽  
James N. Hay
Keyword(s):  
Molecular Weight ◽  
Glass Transition ◽  
Phase Separation ◽  
Liquid Phase ◽  
High Molecular Weight ◽  
Structural Properties ◽  
Thermal History ◽  
Liquid Phase Separation ◽  
High Molecular Weight Poly ◽  
Liquid Liquid Phase Separation

Thermal history and purification effects on the structural properties of PVK were investigated. Liquid–liquid phase separation is suggested to occur by separation of isotactic rich segments from a matrix which is predominantly atactic.

scholarly journals Effects of molecular weight and temperature on liquid–liquid phase separation in particles containing organic species and inorganic salts

2015 ◽  
Vol 15 (3) ◽  
pp. 1351-1365 ◽  
Author(s):  
Y. You ◽  
A. K. Bertram
Keyword(s):  
Molecular Weight ◽  
Phase Separation ◽  
Liquid Phase ◽  
Ammonium Sulfate ◽  
Atmospheric Particles ◽  
Liquid Phase Separation ◽  
Organic Species ◽  
Diffusion Rates ◽  
Measured Liquid ◽  
Liquid Liquid Phase Separation

Abstract. Atmospheric particles containing organic species and inorganic salts may undergo liquid–liquid phase separation when the relative humidity varies between high and low values. To better understand the parameters that affect liquid–liquid phase separation in atmospheric particles, we studied the effects of molecular weight and temperature on liquid–liquid phase separation in particles containing one organic species mixed with either ammonium sulfate or ammonium bisulfate. In the molecular-weight-dependent studies, we measured liquid–liquid phase separation relative humidity (SRH) in particles containing ammonium sulfate and organic species with large molecular weights (up to 1153 Da). These results were combined with recent studies of liquid–liquid phase separation in the literature to assess if molecular weight is a useful parameter for predicting SRH. The combined results, which include results from 33 different particle types, illustrate that SRH does not depend strongly on molecular weight (i.e., a clear relationship between molecular weight and SRH was not observed). In the temperature-dependent studies, we measured liquid–liquid phase separation in particles containing ammonium sulfate mixed with 20 different organic species at 244 ± 1 K, 263 ± 1 K, and 278 ± 1 K; a few particles were also studied at 290 ± 1 K. These new results were combined with previous measurements of the same particle types at 290 ± 1 K. The combined SRH data illustrate that for the organic–ammonium sulfate particles studied, the SRH does not depend strongly on temperature. At most the SRH varied by 9.7% as the temperature varied from 290 to 244 K. The high SRH values (> 65%) in these experiments may explain the lack of temperature dependence. Since water is a plasticizer, high relative humidities can lead to high water contents, low viscosities, and high diffusion rates in the particles. For these cases, unless the temperature is very low, liquid–liquid phase separation is not expected to be kinetically inhibited. The occurrence of liquid–liquid phase separation and SRH did depend strongly on temperature over the range of 290–244 K for particles containing α,4-dihydroxy-3-methoxybenzeneacetic acid mixed with ammonium bisulfate. For this particle type, a combination of low temperatures and low water content likely favored kinetic inhabitation of the liquid–liquid phase separation by slow diffusion rates in highly viscous particles. The combined results suggest that liquid–liquid phase separation is likely a common occurrence in atmospheric particles at temperatures from 244–290 K, although particles that do not undergo liquid–liquid phase separation are also likely common.

scholarly journals Previously uncharacterized interactions between the folded and intrinsically disordered domains impart asymmetric effects on UBQLN2 phase separation

2021 ◽  
Author(s):  
Tongyin Zheng ◽  
Carlos A. Castañeda
Keyword(s):  
Quality Control ◽  
Phase Separation ◽  
Liquid Phase ◽  
Protein Quality ◽  
Intrinsically Disordered ◽  
Intrinsically Disordered Regions ◽  
Uba Domain ◽  
Phase Separation Behavior ◽  
Separation Behavior ◽  
Liquid Liquid Phase Separation

AbstractShuttle protein UBQLN2 functions in protein quality control (PQC) by binding to proteasomal receptors and ubiquitinated substrates via its N-terminal ubiquitin-like (UBL) and C-terminal ubiquitin-associated (UBA) domains, respectively. Between these two folded domains are intrinsically disordered STI1-I and STI1-II regions, connected by disordered linkers. The STI1 regions bind other components, such as HSP70, that are important to the PQC functions of UBQLN2. We recently determined that the STI1-II region enables UBQLN2 to undergo liquid-liquid phase separation (LLPS) to form liquid dropletsin vitroand biomolecular condensates in cells. However, how the interplay between the folded (UBL/UBA) domains and the intrinsically-disordered regions mediates phase separation is largely unknown. Using engineered domain deletion constructs, we found that removing the UBA domain inhibits UBQLN2 LLPS while removing the UBL domain enhances LLPS, suggesting that UBA and UBL domains contribute asymmetrically in modulating UBQLN2 LLPS. To explain these differential effects, we interrogated the interactions that involve the UBA and UBL domains across the entire UBQLN2 molecule using NMR spectroscopy. To our surprise, aside from well-studied canonical UBL:UBA interactions, there also exist moderate and weak interactions between the UBL and STI1-I/STI1-II domains, and between the UBA domain and the linker connecting the two STI1 regions, respectively. Our findings are essential for the understanding of both the molecular driving forces of UBQLN2 LLPS and the effects of ligand binding to UBL, UBA, or STI1 domains on the phase behavior and physiological functions of UBQLN2.Impact of Work StatementZheng and Castañeda show that interplay between the folded domains and intrinsically disordered regions regulates liquid-liquid phase separation behavior of UBQLN2, a protein quality control (PQC) shuttle protein. Despite their similar size, the folded UBL and UBA domains inhibit and promote phase separation, respectively, due to their previously uncharacterized, asymmetric interactions with the middle intrinsically-disordered region. These results strongly suggest that PQC components, including proteasomal receptors, are likely to modulate UBQLN2 phase separation behavior in cells.

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