title: Functional screening of pharmacological chaperones via restoration of enzyme activity upon denaturation.
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.
authors.affiliation: Department of Chemistry and Chemical Biology, McMaster University, Hamilton, ON, Canada L8S 4M1.
authors.forename: Meera
authors.initials: M
authors.lastname: Shanmuganathan
authors.forename: Philip
authors.initials: P
authors.lastname: Britz-McKibbin
mesh_terms: D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004789:Enzyme Activation / Q000187:drug effects*; D005776:Gaucher Disease / Q000188:drug therapy / Q000201:enzymology; D005962:Glucosylceramidase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding / Q000187:drug effects*; D055550:Protein Stability / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D008024:Ligands; D054852:Small Molecule Libraries; D005962:Glucosylceramidase