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bioRxiv · 10.1101/563700

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

van Lindt, J.·Bratek-Skicki, A.·Pakravan, D.·Van Den Bosch, L.·Maes, D.·Tompa, P.
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Abstract

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.

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van Lindt, J., Bratek-Skicki, A., Pakravan, D., Van Den Bosch, L., Maes, D., Tompa, P.. 2019-02-28. A generic approach for studying the kinetics of liquid-liquid phase separation under near-native conditions. https://doi.org/10.1101/563700

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