A generic approach to study the kinetics of liquid-liquid phase separation under near-native conditions.
Commun Biol
; 4(1): 77, 2021 01 19.
Article
de En
| MEDLINE
| ID: mdl-33469149
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.
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Protéines de liaison à l'ADN
/
Extraction liquide-liquide
Langue:
En
Journal:
Commun Biol
Année:
2021
Type de document:
Article
Pays d'affiliation:
Belgique