DiscussionPriyanka Narasimhan, Hayley Angove, Stephanie Beaver, Katie Chapman,
Marissa Flower, Stuart Firth-Clark, Gary Newton, Trevor Perrior,
Jim Reid, Debbie Sanders, Mark Stewart
Domainex Ltd., 162 Cambridge Science Park, CB4 0GH Cambridge, UK
Lysine methyltransferases (KMTs) are involved in epigenetic gene regulation by covalent modification of histones. In particular, a number of these enzymes are reported to be of relevance in some mechanisms of carcinogenesis. Domainex has addressed key technical drug discovery challenges associated with KMTs, including generating a number of proprietary crystal structures and assays. In particular, an EZH2 screening assay was developed using both a peptide substrate and more physiologically relevant substrate H3/H4 tetramer. Biochemical assays were then utilised to screen a PKMT library identified using our LeadBuilder virtual screening technology. EZH2 specific chemotypes were resolved over SMYD2, SMYD3, and G9a. EZH2 is known to produce the histone H3K27(me3) modification, and we have demonstrated cellular efficacy of tool compounds by measuring histone methylation via both FACS and Western blotting techniques.