Open AccessRapid Evaluation of Small Molecule Cellular Target Engagement with a Luminescent Thermal Shift Assay
Author(s) -
Jonathan D. Mortison,
Iván CornellaTaracido,
Gireedhar Venkatchalam,
Anthony W. Partridge,
Nirodhini S Siriwardana,
Simon M. Bushell
Publication year - 2021
Publication title -
acs medicinal chemistry letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.065
H-Index - 66
ISSN - 1948-5875
DOI - 10.1021/acsmedchemlett.1c00276
Subject(s) - small molecule , drug discovery , molecule , chemistry , luminescence , computational biology , materials science , biology , biochemistry , optoelectronics , organic chemistry
Determination of target engagement for candidate drug molecules in the native cellular environment is a significant challenge for drug discovery programs. The cellular thermal shift assay (CETSA) has emerged as a powerful tool for determining compound target engagement through measurement of changes to a protein's thermal stability upon ligand binding. Here, we present a HiBiT thermal shift assay (BiTSA) that deploys a quantitative peptide tag for determination of compound target engagement in the native cellular environment using a high throughput, plate-based luminescence readout. We demonstrate that BiTSA can rapidly assess cellular target engagement of small molecule ligands against their cognate targets and highlight two applications of BiTSA for differentiating small molecules targeting mutant KRAS and TP53.