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- W3132392321 abstract "Compartmentalization by liquid-liquid phase separation (LLPS) has emerged as a unifying theme in biology and as a mechanism for organizing biomolecules in space and time. LLPS can provide a pathway for cells to rapidly respond to changes in their environment. Under biologically relevant conditions, LLPS likely proceeds through nucleation-and-growth and the rate at which phase separated compartments can form is determined by the nucleation barrier. To characterize the nature of the energetic barrier to nucleation, we have combined equilibrium techniques that define the phase boundaries of a prototypical prion-like domain and the size distribution of clusters below the nucleation barrier with rapid mixing time-resolved small-angle x-ray scattering (SAXS). SAXS is sensitive to structures on the nanoscale and rapid mixing microfluidics allow us to probe the size evolution of the system as it progresses toward two-phase equilibrium. We demonstrate that phase separation of prion-like domains can be described by homogeneous classical nucleation theory on length scales greater than small oligomers. However, this theory breaks down for small oligomers where an additional energy barrier to assembly exists. These results suggest that a conformational change is required for nucleation and present an experimental methodology by which nucleation can be rigorously studied." @default.
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- W3132392321 date "2021-02-01" @default.
- W3132392321 modified "2023-09-27" @default.
- W3132392321 title "Kinetic and Equilibrium Measurement of Nucleation of Prion-Like Domain Phase Separation" @default.
- W3132392321 doi "https://doi.org/10.1016/j.bpj.2020.11.871" @default.
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