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

• DOI: 10.1038/s42003-020-01596-8
• Publication Date: 2021-01-19T11:02:54Z
• Authors: Joris Van Lindt; Anna Bratek-Skicki; Phuong N. Nguyen; Donya Pakravan; Luis F. Durán-Arm...; Agnes Tantos; Rita Pancsa; Ludo Van Den Bosch; Dominique Maes; Peter Tompa
• Keywords: DNA-Binding Proteins; 0301 basic medicine; Kinetics; 0303 health sciences; 03 medical and health sciences; Protein Domains; QH301-705.5; Liquid-Liquid Extraction; Thermodynamics; Biology (General); Article; Biophysical Phenomena; 3. Good health

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