Target identification using drug affinity responsive target stability (DARTS)

TLDR

Identifying the molecular targets of small‑molecule drugs remains an important and unmet challenge. The authors developed DARTS, a method that exploits reduced protease susceptibility of drug‑bound proteins and propose it for global mapping of protein–metabolite interactions and label‑free screening of diverse compounds for imaging agents. DARTS detects drug–target interactions by measuring decreased protease susceptibility of the bound protein, requiring no drug modification and functioning regardless of the drug’s mechanism of action. Using DARTS, the authors identified known drug–protein interactions and discovered that resveratrol targets the eukaryotic translation initiation machinery, a potential longevity pathway.

Abstract

Identifying the molecular targets for the beneficial or detrimental effects of small-molecule drugs is an important and currently unmet challenge. We have developed a method, drug affinity responsive target stability (DARTS), which takes advantage of a reduction in the protease susceptibility of the target protein upon drug binding. DARTS is universally applicable because it requires no modification of the drug and is independent of the mechanism of drug action. We demonstrate use of DARTS to identify known small-molecule-protein interactions and to reveal the eukaryotic translation initiation machinery as a molecular target for the longevity-enhancing plant natural product resveratrol. We envisage that DARTS will also be useful in global mapping of protein-metabolite interaction networks and in label-free screening of unlimited varieties of compounds for development as molecular imaging agents.