Assessing target engagement using proteome-wide solvent shift assays.

Area of Science:

• Biochemistry
• Proteomics
• Drug Discovery

Background:

• Quantitative proteomics using mass spectrometry is vital for drug discovery.
• Proteins unfold and precipitate in organic solvents, a process influenced by ligand binding.
• Understanding protein-ligand interactions is key to identifying drug targets.

Purpose of the Study:

• To develop novel proteome-wide assays for target engagement studies in drug discovery.
• To introduce solvent proteome profiling (SPP) and solvent proteome integral solubility alteration (solvent-PISA) as efficient methods.

Main Methods:

• Combined solvent-induced precipitation principles with quantitative mass spectrometry.
• Developed solvent proteome profiling (SPP) to analyze protein denaturation curves.
• Applied area under the curve analysis to create solvent-PISA as a surrogate for SPP.

Main Results:

• Successfully identified specific targets of compounds with known mechanisms of action.
• Demonstrated that solvent-PISA reliably serves as a surrogate for SPP.
• Established SPP's capability in establishing target engagement.

Conclusions:

• SPP and solvent-PISA offer efficient proteome-wide screening for drug target engagement.
• Combining SPP with methods like thermal proteome profiling can increase the number of high-quality binding assays.
• These approaches expand the fraction of the proteome that can be screened for ligand binding, accelerating drug discovery.