Drug Affinity Responsive Target Stability (DARTS) to Resolve Protein-Small Molecule Interaction in Arabidopsis

Area of Science:

• Plant biology
• Chemical genomics
• Biochemistry

Background:

• Identifying small molecule targets in plants is crucial for chemical genomics.
• Existing methods may require molecule modification or are mechanism-dependent.

Purpose of the Study:

• To detail the Drug Affinity Responsive Target Stability (DARTS) technique for plant protein targets.
• To provide adaptable detection protocols for varying protein abundance levels.

Main Methods:

• Application of the DARTS protocol to plant protein extracts.
• Treatment of protein extracts with proteolytic enzymes.
• Protection of small molecule-bound proteins from proteolysis.

Main Results:

• DARTS effectively identifies protein targets for small molecules in plants.
• The method is independent of the small molecule's mechanism or structure.
• No additional molecule modification or tagging is required.

Conclusions:

• DARTS is a versatile and straightforward technique for plant target identification.
• The described protocols facilitate the characterization of small molecule-protein interactions.
• This method supports advancements in plant chemical genomics.