Biophysical characterization of membrane protein-small molecule interactions

Membrane proteins are account for up to two thirds of known druggable targets. Traditionally, new drugs against this class of proteins have been discovered through HTS. However, not all GPCRs are amenable to traditional screening methods. Recently, fragment-based drug discovery (FBDD) has emerged as a powerful strategy to generate approved drugs against soluble targets. Now, FBDD can be applied to GPCRs with great potential advantages. In recent years, a number of FBDD techniques have been validated for use with GPCRs. The impressive growth in GPCR structure information leads to broad use of structure-based methods for hit discovery and optimization. However, the dynamic nature of GPCRs, and standard issues associated with low level expression and instability during purification, made biophysical and structural characterization of GPCRs particularly difficult. New advances in protein stabilization by using different protein engineering methods and alternative solubilization strategy have shown the potential to facilitate GPCR structural and biophysical studies. The goal of the work described in this thesis was to develop and implement efficient fragment screening methods to discover ligands of GPCRs with novel biological activities, and new advances in receptor production and stabilization to facilitate structural biology of GPCRs in the early stages of drug discovery.UBL - phd migration 201