A Method for Conditional Regulation of Protein Stability in Native or Near-Native Form

Cell Chem Biol. 2020 Dec 17;27(12):1573-1581.e3. doi: 10.1016/j.chembiol.2020.09.004. Epub 2020 Oct 1.


Here, we report a method to regulate cellular protein levels by introducing a ubiquitin variant between a destabilizing domain (DD) and the regulated protein. When produced in the absence of a stabilizing ligand the DD dominates and the entire fusion protein is processively degraded by the proteasome. In the presence of the stabilizing ligand the fusion protein is metabolically stable and becomes a substrate for abundant ubiquitin-specific proteases, liberating a native, or a near-native protein-of-interest. This technique is thus particularly useful for the study of proteins whose free N terminus is required for proper function. In addition, removal of the DD in the presence of stabilizing ligand leads to higher expression levels of regulated protein when cells experience transient exposure to a stabilizing ligand, such as in a living animal receiving a single dose of a pharmacological agent as the stabilizing ligand.

Keywords: degradation; destabilized domain; deubiquitinase; protein stability; ubiquitin.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Peptide Hydrolases / metabolism
  • Protein Domains
  • Protein Engineering / methods*
  • Protein Stability
  • Recombinant Fusion Proteins / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Ubiquitin / chemistry
  • Ubiquitin / genetics
  • Ubiquitin / metabolism*


  • Recombinant Fusion Proteins
  • Ubiquitin
  • Peptide Hydrolases