Evaluation of Cooling-Induced Liquid-Liquid Phase Separation of Ureido Polymers as a Cold-Shock Stress Granules Model

Macromol Biosci. 2021 Mar;21(3):e2000345. doi: 10.1002/mabi.202000345. Epub 2021 Jan 15.

Abstract

Many intracellular reactions occur in membrane-less organelles that form due to liquid-liquid phase separation (LLPS). Cold-shock stress granules, which are membrane-less organelles, are formed in response to a significant decrease in temperature and recruit biomolecules for regulation of their activities. The authors have reported that synthetic ureido copolymers exhibit cooling-induced LLPS under physiologically relevant conditions. In this study, influences of the cooling-induced LLPS of ureido polymers on enzymatic activity is investigated to evaluate whether the ureido polymers can mimic cold-shock stress granules. The enzyme β-galactosidase (β-Gal) is efficiently entrapped into phase-separated coacervates of ureido polymers upon cooling. The activity of β-Gal is significantly suppressed by the entrapment. The enzymatic activity is recovered after heating, which dissolves the coacervate. Thus, the LLPS formed by ureido polymers are a suitable model for cold-shock stress granules.

Keywords: cold-shock stress granules; liquid-liquid phase separation; membrane-less organelles; upper critical solution temperature; ureido copolymers.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cattle
  • Cold Temperature*
  • Cold-Shock Response*
  • Cytoplasmic Granules / metabolism*
  • Fluorescein-5-isothiocyanate / metabolism
  • Kinetics
  • Liquid-Liquid Extraction*
  • Polymers / chemistry*
  • Urea / chemistry*
  • beta-Galactosidase / metabolism

Substances

  • Polymers
  • Urea
  • beta-Galactosidase
  • Fluorescein-5-isothiocyanate