Development of a target identification approach using native mass spectrometry.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Pharmacology

Background:

• Identifying molecular targets is critical for pharmaceutical drug development.
• Current target identification methods are a bottleneck in advancing drug discovery.
• Distinguishing direct drug targets from indirect responders is essential.

Purpose of the Study:

• To report a novel method for identifying molecular targets using native mass spectrometry.
• To demonstrate the detection of protein-ligand complexes for target identification.
• To overcome limitations of requiring labeled ligands in drug discovery.

Main Methods:

• Utilized native mass spectrometry (MS) for specific detection of protein-ligand complexes.
• Employed unmodified small molecules for binding assays.
• Tested the method in protein mixtures and bacterial cell lysates.

Main Results:

• Successfully detected known ligand-protein interactions in complex biological samples.
• Identified the protein-ligand complex between parthenolide and thioredoxin (PfTrx).
• Demonstrated detection in a five-protein mixture and a spiked bacterial cell lysate.

Conclusions:

• Native MS is a powerful, untargeted method for identifying molecular targets.
• The method does not require chemical modification or labeling of small molecules.
• Native MS shows potential for broad application in identifying binding targets for diverse small molecules.