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Proc Natl Acad Sci U S A. 2014 Sep 2;111(35):12853-8. doi: 10.1073/pnas.1407358111. Epub 2014 Aug 18.

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

Author information

1
Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; carrow@uhnres.utoronto.ca m.vedadi@utoronto.ca d.barsyte@utoronto.ca.
2
Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7;
3
Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada V6T1Z3;
4
Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, San Diego, CA 92121;
5
Department of Microbiology and Immunology, University of British Columbia, Vancouver, BC, Canada V6T1Z3;
6
Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada M5S 1A8;
7
Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, ON, Canada M5G 0A3;
8
Centre for Systems Biology, Lunenfeld-Tanenbaum Research Institute, Toronto, ON, Canada M5G 1X5;
9
Centre for Systems Biology, Lunenfeld-Tanenbaum Research Institute, Toronto, ON, Canada M5G 1X5; Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada M5S 1A8;
10
Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada M5S 1A8; Drug Discovery Program, Ontario Institute for Cancer Research, Toronto, ON, Canada M5G 0A3;
11
Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, Sandwich, Kent CT13 9NJ, United Kingdom;
12
Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, Groton, CT 06340;
13
Worldwide Medicinal Chemistry, Pfizer Worldwide Research and Development, Cambridge, MA 02139;
14
Neusentis Research Unit, Pfizer Worldwide Research and Development, Cambridge CB21 6GS, United Kingdom;
15
Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada V6T1Z3; Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada V6T1Z3;
16
Weis Center for Research, Geisinger Clinic, Danville, PA 17821;
17
Biomedical Research Centre, University of British Columbia, Vancouver, BC, Canada V6T1Z3; Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada V6T1Z3; and.
18
Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; Department of Pharmacology and Toxicology, University of Toronto, Toronto, ON, Canada M5S 1A8; carrow@uhnres.utoronto.ca m.vedadi@utoronto.ca d.barsyte@utoronto.ca.
19
Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada M5G 1L7; Princess Margaret Cancer Centre and Department of Medical Biophysics, University of Toronto, ON, Canada M5G 1L7 carrow@uhnres.utoronto.ca m.vedadi@utoronto.ca d.barsyte@utoronto.ca.

Abstract

SET domain containing (lysine methyltransferase) 7 (SETD7) is implicated in multiple 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 (Ki (app) = 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.

KEYWORDS:

chemical biology; chemical probe; epigenetics

PMID:
25136132
PMCID:
PMC4156762
DOI:
10.1073/pnas.1407358111
[Indexed for MEDLINE]
Free PMC Article

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