(R)-PFI-2 is a potent and selective inhibitor of SETD7 methyltransferase activity in cells

Dalia Barsyte-Lovejoy, Fengling Li, Menno J. Oudhoff, John H. Tatlock, Aiping Dong, Hong Zeng, Hong Wu, Spencer A. Freeman, Matthieu Schapira, Guillermo A. Senisterra, Ekaterina Kuznetsova, Richard Marcellus, Abdellah Allali-Hassani, Steven Kennedy, Jean Philippe Lambert, Amber L. Couzens, Ahmed Aman, Anne Claude Gingras, Rima Al-Awar, Paul V. FishBrian S. Gerstenberger, Lee Roberts, Caroline L. Benn, Rachel L. Grimley, Mitchell J.S. Braam, Fabio M.V. Rossi, Marius Sudol, Peter J. Brown, Mark E. Bunnage, Dafydd R. Owen, Colby Zaph, Masoud Vedadi, Cheryl H. Arrowsmith

Research output: Contribution to journalArticlepeer-review

142 Scopus citations


SET domain containing (lysine methyltransferase) 7 (SETD7) is implicated inmultiple signaling and disease related pathways with a broad diversity of reported substrates. Here, we report the discovery of (R)-PFI-2 - a first-in-class, potent (Kiapp = 0.33 nM), selective, and cell-active inhibitor of the methyltransferase activity of human SETD7 - and its 500-fold less active enantiomer, (S)-PFI-2. (R)-PFI-2 exhibits an unusual cofactor-dependent and substrate-competitive inhibitory mechanism by occupying the substrate peptide binding groove of SETD7, including the catalytic lysine-binding channel, and by making direct contact with the donor methyl group of the cofactor, S-adenosylmethionine. Chemoproteomics experiments using a biotinylated derivative of (R)-PFI-2 demonstrated dose-dependent competition for binding to endogenous SETD7 in MCF7 cells pretreated with (R)-PFI-2. In murine embryonic fibroblasts, (R)-PFI-2 treatment phenocopied the effects of Setd7 deficiency on Hippo pathway signaling, via modulation of the transcriptional coactivator Yes-associated protein (YAP) and regulation of YAP target genes. In confluent MCF7 cells, (R)-PFI-2 rapidly altered YAP localization, suggesting continuous and dynamic regulation of YAP by the methyltransferase activity of SETD7. These data establish (R)-PFI-2 and related compounds as a valuable tool-kit for the study of the diverse roles of SETD7 in cells and further validate protein methyltransferases as a druggable target class.

Original languageEnglish
Pages (from-to)12853-12858
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number35
StatePublished - 2 Sep 2014
Externally publishedYes


  • Chemical biology
  • Chemical probe
  • Epigenetics


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