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Liquid-liquid phase separation (LLPS) of proteins into concentrated microdroplets (also called coacervation) is a phenomenon that is increasingly recognized to occur in many biological processes, both inside and outside the cell. While it has been established that LLPS can be described as a spinodal decomposition leading to demixing of an initially homogenous protein solution, little is known about the assembly pathways by which soluble proteins aggregate into dense microdroplets. Using a recently developed technique enabling the observation of matter suspended in liquid by transmission electron microscopy (TEM), we observed how a model intrinsically disordered protein (IDP) phase-separates in liquid environment. Our observations reveal for the first time dynamic mechanisms by which soluble proteins self-organize into condensed microdroplets with nano-scale and milli-second space and time resolution, respectively. With this method, the nucleation and initial growth steps of LLPS could be captured, opening the door for a deeper understanding of biomacromolecular complexes exhibiting LLPS ability.
This article was published in the following journal.
Name: Journal of the American Chemical Society
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Miniaturized methods of liquid-liquid extraction.
A method of separation of two or more substances by repeated distribution between two immiscible liquid phases that move past each other in opposite directions. It is a form of liquid-liquid chromatography. (Stedman, 25th ed)
Fractionation of a vaporized sample as a consequence of partition between a mobile gaseous phase and a stationary phase held in a column. Two types are gas-solid chromatography, where the fixed phase is a solid, and gas-liquid, in which the stationary phase is a nonvolatile liquid supported on an inert solid matrix.
The removal of a soluble component from a liquid mixture by contact with a second liquid, immiscible with the carrier liquid, in which the component is preferentially soluble. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)
Chromatographic techniques in which the mobile phase is a liquid.
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