Trapping an enzyme to map its network of influence within the cell

The protocol incorporates an in silico validation phase (Phase 1) with explicit GO/NO-GO criteria, which permits the filtering of targets and conditions before commitment to costly experiments — a best practice in sequential experimental design.

The hypothesis convincingly exploits the mechanistic specificity of ABPs (activity-dependent, irreversible inhibition) to create a temporally precise perturbation, which is a clear advantage over reversible inhibitors or genetic perturbations that have longer and less controllable delays in action.

The idea of using an ABP probe as a temporally precise causal intervention is conceptually elegant and addresses a genuine methodological need in network inference.

Captive and solvent market: large pharmaceutical companies (Novartis, Pfizer, Roche) and biotechs in oncology/neurodegeneration (e.g., Alnylam for DUBs) would pay to validate orphan enzymatic targets or to deconvolute mechanisms of resistance to inhibitors, with a typical R&D budget of €500k–2M per target validation project.

A mechanistically elegant and disruptive hypothesis is proposed: the use of ABPs as causal perturbagens for network inference, moving beyond simple imaging or reversible inhibition.