TY - JOUR
T1 - A generic approach to study the kinetics of liquid–liquid phase separation under near-native conditions
AU - Lindt, Joris Van
AU - Bratek-Skicki, Anna
AU - Nguyen, Phuong N.
AU - Pakravan, Donya
AU - Durán-Armenta, Luis F.
AU - Tantos, Agnes
AU - Pancsa, Rita
AU - Bosch, Ludo Van Den
AU - Maes, Dominique
AU - Tompa, Peter
PY - 2021/1/19
Y1 - 2021/1/19
N2 - 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.
AB - 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.
UR - https://doi.org/10.1038/s42003-020-01596-8
UR - http://www.scopus.com/inward/record.url?scp=85099540323&partnerID=8YFLogxK
U2 - 10.1038/s42003-020-01596-8
DO - 10.1038/s42003-020-01596-8
M3 - Article
C2 - 33469149
VL - 4
JO - Communications Biology
JF - Communications Biology
SN - 2399-3642
IS - 1
M1 - 77
ER -