Osmolytes Can Destabilize Proteins in Cells by Modulating Electrostatics and Quinary Interactions

Although numerous in vitro studies have shown that osmolytes are capable of stabilizing proteins, their effect on protein folding in vivo has been less understood. In this work, we investigated the effect of osmolytes, including glycerol, sorbitol, betaine, and taurine, on the folding of a protein GB3 variant in E. coli cells using NMR spectroscopy. 400 mM osmolytes were added to E. coli cells; only glycerol stabilizes the folded protein, whereas betaine and taurine considerably destabilize the protein through modulating folding and unfolding rates. Further investigation indicates that betaine and taurine can enhance the quinary interaction between the protein and cellular environment and manifestly weaken the electrostatic attraction in protein salt bridges. The combination of the two factors causes destabilization of the protein in E. coli cells. These factors counteract the preferential exclusion mechanism that is adopted by osmolytes to stabilize proteins.

Automated summary

This study found that glycerol stabilizes protein folding, while betaine and taurine destabilize protein folding by affecting folding rates and instability. Betaine and taurine enhance quinary interactions between the protein and cellular environment, weakening electrostatic attraction in protein salt bridges, leading to protein destabilization in E. coli cells. These factors counteract the preferential exclusion mechanism used by osmolytes to stabilize proteins.