Liquid-Liquid Phase Separation to Study the Association of Proteins in Solution

Methods Mol Biol. 2023:2551:253-267. doi: 10.1007/978-1-0716-2597-2_17.

Abstract

Liquid-liquid phase separation (LLPS) is a reversible biological process that contributes to the formation of critical concentration of proteins, forming membraneless compartments that are physiologically and pathologically relevant. Several proteins have been shown to demix into liquid droplets under in vitro crowding conditions. These studies are mainly conducted in isolation using purified recombinant proteins. Recently, we used LLPS to study the association between two proteins that are co-aggregated in Alzheimer's disease brain, tau, and EFhd2. Here, we describe how we used LLPS to determine the molecular components that contribute to the transition of these two proteins from liquid droplets to solid-like structures.

Keywords: Aggregates; EFhd2; Liquid-liquid phase separation; Protein dynamics; Tau.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease* / metabolism
  • Brain / metabolism
  • Humans
  • Recombinant Proteins / metabolism
  • tau Proteins / metabolism

Substances

  • Recombinant Proteins
  • tau Proteins