Rescue of misfolded proteins and stabilization by small molecules

Increasing stability of functional proteins by binding small compounds and ions has long been used to extend shelf-life of protein formulations in the pharmacological and biotechnological industry. Likewise, the therapeutic application of small molecules for in vivo recovery and maintenance of structure and function of proteins is steadily increasing. Compounds that can rescue misfolded proteins by stimulating their correct folding and/or the stabilization of native-like conformations in vivo are referred to as pharmacological chaperones. Here we present thermal-shift and isothermal methods for the high-throughput screening of stabilizing pharmacological chaperones for soluble and membrane proteins. The effect of selected hit compounds on the kinetics of protein synthesis is further evaluated by an in vitro transcription-translation rapid translation system. These procedures can be integrated in an interdisciplinary and translational approach for the search of personalized pharmacological chaperones in genetic misfolding diseases.