BIX-01294 and its analogs were originally identified and subsequently designed as potent inhibitors against histone H3 lysine 9 (H3K9) methyltransferases G9a and G9a-like protein. Here, we show that BIX-01294 and its analog E67 can also inhibit H3K9 Jumonji demethylase KIAA1718 with half-maximal inhibitory concentrations in low micromolar range. Crystallographic analysis of KIAA1718 Jumonji domain in complex with E67 indicated that the benzylated six-membered piperidine ring was disordered and exposed to solvent. Removing the moiety (generating compound E67-2) has no effect on the potency against KIAA1718 but, unexpectedly, lost inhibition against G9a-like protein by a factor of 1500. Furthermore, E67 and E67-2 have no effect on the activity against histone H3 lysine 4 (H3K4) demethylase JARID1C. Thus, our study provides a new avenue for designing and improving the potency and selectivity of inhibitors against H3K9 Jumonji demethylases over H3K9 methyltransferases and H3K4 demethylases.
Bibliographical noteFunding Information:
We thank Dr. John R. Horton for maintaining local X-ray facility and help with X-ray data collection in synchrotron, Dr. Justin E. Jones for comments and Dr. Robert A. Copeland for suggestion of calculating K i app values. The Department of Biochemistry at the Emory University School of Medicine supported the use of the Southeast Regional Collaborative Access Team synchrotron beamline at the Advanced Photon Source of Argonne National Laboratory, local X-ray facility and matrix-assisted laser desorption/ionization–time of flight mass spectrometry. This work was supported by National Institutes of Health grants ( GM068680-07 to X.C. and GM092035-01 to X.C. and Y.Y.), by PRIN 2009PX2T2E and by the FP7 Project BLUEPRINT/282510 to A.M. X.C. is a Georgia Research Alliance Eminent Scholar.
All Science Journal Classification (ASJC) codes
- Structural Biology
- Molecular Biology