Target Discovery Using Thermal Proteome Profiling
Sindhuja Sridharan, Ina Günthner, Isabelle Becher, Mikhail M. Savitski, Marcus Bantscheff · 2019
The drug discovery process involves the search for prospective leads from small-molecule libraries by using either target-based assays or cell-based, phenotypic assays. Ligand binding alters both biochemical and biophysical properties of a target protein, which may also affect its interactions with other proteins and metabolites in different cellular components. Thermal proteome profiling (TPP) measures the heat-induced denaturation of proteins and identifies drug-bound targets based on altered thermal stability. The chapter discusses the different experimental formats of TPP and the data analysis strategies that have been developed for different applications and experimental schemes. The combination of cellular thermal shift assay (CETSA) with quantitative proteomics transcended a target engagement tool to a TPP approach that enables the comprehensive discovery of drug protein interactions in live cell systems. The primary goal of CETSA has been to measure drug-target interactions inside cells.