Figure 1.

Design and synthesis of dual‐tagged ezutromid probes for use in affinity‐based protein profiling. a) Outline of the AfBPP strategy for target identification of small molecule probes in live cells. Probe‐treated cells are UV irradiated to trigger covalent binding of the probe to its protein target. A click reaction is performed to ligate a fluorophore or enrichment tag, such as biotin, to the probe. Enriched proteins are subsequently analysed using western blot or mass spectrometry. b) Chemical structures of ezutromid (1) ezutromid‐based active (2–3) and inactive (4) probes, containing a diazirine as a photoaffinity label (yellow shading) and alkyne click handles (green shading). c) Synthesis of probes 2 and 3.