Early Stage Discovery and Validation of Pharmacological Chaperones for the Correction of Protein Misfolding Diseases

Methods Mol Biol. 2019;1873:279-292. doi: 10.1007/978-1-4939-8820-4_18.


Pharmacological chaperones are small molecular weight molecules that bind specifically to protein targets and stabilize unstable and misfolded conformations. In particular, there is an increasing interest in the application of this type of compounds for the correction of genetic conformational disorders, which are caused by mutations leading to protein instability. The discovery of compounds with pharmacological chaperone ability is customarily initiated by a high-throughput screening of chemical libraries searching for stabilizing binders. However, there is no established consensus for the subsequent steps. Therefore, here, we introduce an example of a successful protocol directed to the discovery of pharmacological chaperones with potential for the therapeutic correction of phenylketonuria, a defect caused by mutations in the enzyme phenylalanine hydroxylase.

Keywords: Drug discovery and development; Functional assays; Orthogonal binding assays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques
  • Cell Line
  • Drug Discovery* / methods
  • Enzyme Activation / drug effects
  • Gene Expression
  • Genes, Reporter
  • High-Throughput Screening Assays
  • Humans
  • Ligands
  • Molecular Chaperones / chemistry*
  • Molecular Chaperones / metabolism
  • Phenylalanine Hydroxylase / antagonists & inhibitors
  • Phenylalanine Hydroxylase / chemistry
  • Protein Folding* / drug effects
  • Proteostasis Deficiencies / drug therapy
  • Small Molecule Libraries
  • Surface Plasmon Resonance / methods


  • Ligands
  • Molecular Chaperones
  • Small Molecule Libraries
  • Phenylalanine Hydroxylase