A chemical proteomic analysis of illudin-interacting proteins

The illudin natural product family are fungal secondary metabolites with a characteristic spirocyclopropyl-substituted fused 6,5-bicyclic ring system. They have been extensively studied for their cytotoxicity in various tumor cell types and semisynthetic derivatives with improved therapeutic characteristics have progressed into clinical trials. While it is believed that this potent alkylating compound class is mainly acting via DNA modification, little is known about its binding to protein sites in a cellular context. In order to reveal putative protein targets of the illudin family in live cancer cells, we employed a semisynthetic strategy to access a series of illudin-based probes for activity-based protein profiling (ABPP). While the probes largely retained potent cytotoxicity, proteomic profiling studies unravelled multiple protein hits suggesting that illudins exert their mode of action not from addressing a specific protein target but rather from DNA modification and unselective protein binding.