A generic approach to study the kinetics of liquid–liquid phase separation under near-native conditions

Joris Van Lindt, Anna Bratek-Skicki, Phuong N. Nguyen, Donya Pakravan, Luis F. Durán-Armenta, Agnes Tantos, Rita Pancsa, Ludo Van Den Bosch, Dominique Maes, Peter Tompa

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1 Citation (Scopus)

Abstract

Understanding the kinetics, thermodynamics, and molecular mechanisms of liquid–liquid phase separation (LLPS) is of paramount importance in cell biology, requiring reproducible methods for studying often severely aggregation-prone proteins. Frequently applied approaches for inducing LLPS, such as dilution of the protein from an urea-containing solution or cleavage of its fused solubility tag, often lead to very different kinetic behaviors. Here we demonstrate that at carefully selected pH values proteins such as the low-complexity domain of hnRNPA2, TDP-43, and NUP98, or the stress protein ERD14, can be kept in solution and their LLPS can then be induced by a jump to native pH. This approach represents a generic method for studying the full kinetic trajectory of LLPS under near native conditions that can be easily controlled, providing a platform for the characterization of physiologically relevant phase-separation behavior of diverse proteins.
Original languageEnglish
Article number77
Number of pages8
JournalCommunications Biology
Volume4
Issue number1
DOIs
Publication statusPublished - 19 Jan 2021

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