Cell Volume Controls Protein Stability and Compactness of the Unfolded State

J Phys Chem B. 2018 Dec 13;122(49):11762-11770. doi: 10.1021/acs.jpcb.8b08216. Epub 2018 Oct 26.

Abstract

Macromolecular crowding is widely accepted as one of the factors that can alter protein stability, structure, and function inside cells. Less often considered is that crowding can be dynamic: as cell volume changes, either as a result of external duress or in the course of the cell cycle, water moves in or out through membrane channels, and crowding changes in tune. Both theory and in vitro experiments predict that protein stability will be altered as a result of crowding changes. However, it is unclear how much the structural ensemble is altered as crowding changes in the cell. To test this, we look at the response of a FRET-labeled kinase to osmotically induced volume changes in live cells. We examine both the folded and unfolded states of the kinase by changing the temperature of the media surrounding the cell. Our data reveals that crowding compacts the structure of its unfolded ensemble but stabilizes the folded protein. We propose that the structure of proteins lacking a rigid, well-defined tertiary structure could be highly sensitive to both increases and decreases in cell volume. Our findings present a possible mechanism for disordered proteins to act as sensors and actuators of cell cycle or external stress events that coincide with a change in macromolecular crowding.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Calibration
  • Cell Line, Tumor
  • Cell Size*
  • Fluorescence Resonance Energy Transfer
  • Humans
  • Macromolecular Substances / chemistry
  • Osmotic Pressure
  • Protein Stability*
  • Protein Unfolding*
  • Proteins / chemistry*
  • Temperature

Substances

  • Macromolecular Substances
  • Proteins