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

Onderzoeksoutput: Articlepeer review

28 Citaten (Scopus)

Samenvatting

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.
Originele taal-2English
Artikelnummer77
Aantal pagina's8
TijdschriftCommunications Biology
Volume4
Nummer van het tijdschrift1
DOI's
StatusPublished - 19 jan 2021

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