Functional screening of pharmacological chaperones via restoration of enzyme activity upon denaturation

Biochemistry. 2012 Oct 2;51(39):7651-3. doi: 10.1021/bi301223f. Epub 2012 Sep 19.

Abstract

Pharmacological chaperones (PCs) are small molecules that stabilize and promote protein folding. Enzyme inhibition is widely used for PC selection; however, it does not accurately reflect chaperone activity. We introduce a functional assay for characterization of PCs based on their capacity to restore enzyme activity that is abolished upon chemical denaturation. Dose-dependent activity curves were performed as a function of urea to assess the chaperone potency of various ligands to β-glucocerebrosidase as a model system. Restoration of enzyme activity upon denaturation allows direct screening of PCs for treatment of genetic disorders associated with protein deficiency, such as Gaucher disease.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Drug Evaluation, Preclinical / methods*
  • Enzyme Activation / drug effects*
  • Gaucher Disease / drug therapy
  • Gaucher Disease / enzymology
  • Glucosylceramidase / chemistry
  • Glucosylceramidase / metabolism*
  • Humans
  • Ligands
  • Protein Denaturation / drug effects
  • Protein Folding / drug effects*
  • Protein Stability / drug effects*
  • Small Molecule Libraries / chemistry
  • Small Molecule Libraries / pharmacology*

Substances

  • Ligands
  • Small Molecule Libraries
  • Glucosylceramidase