Probe Summary for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)
The fine interplay among methylation states of several lysine residues on the tails of histone proteins is a major determinant of the transcriptional state of the associated DNA coding regions and is commonly referred to as the histone code. Histone lysine demethylases catalyze the removal of methyl groups from methylated lysine sidechains on histones H3 and H4, thus antagonizing the reactions more ..
The fine interplay among methylation states of several lysine residues on the tails of histone proteins is a major determinant of the transcriptional state of the associated DNA coding regions and is commonly referred to as the histone code. Histone lysine demethylases catalyze the removal of methyl groups from methylated lysine sidechains on histones H3 and H4, thus antagonizing the reactions catalyzed by histone lysine methyltransferases. The quest to define the biological roles of the multiple epigenetic modulator enzymes includes the identification and use of small molecules that selectively inhibit individual histone-modifying enzymes/enzyme subfamilies. In search for novel inhibitors of JMJD2E demethylase, a member of the largest set of histone demethylases belonging to the Fe(II) and 2-oxoglutarate oxygenase (2OG) superfamily, we performed a quantitative high-throughput screen (Inglese 2006) by using an assay which utilizes a trimethylated peptide substrate corresponding to a fragment of histone H3 (sequence ARKme3STGGK) with detection of the formaldehyde co-product in real time by a formaldehyde dehydrogenase (FDH) coupled reaction. FDH catalyzes oxidation of formaldehyde to formic acid with the concomitant reduction of the non-fluorescent beta-nicotinamide adenine dinucleotide hydrate (NAD+) cofactor to the fluorescent NADH co-product.
A particular goal in this project is to co-crystalize a small molecule probe with JMJD2E.
This assay will summarize the probe development and crystallization efforts that are currently ongoing.
Inglese, J., Auld, D., Jadhav, A., Johnson, R., Simeonov, A., Yasgar, A., Zheng, W. and Austin, C. Proc Natl Acad Sci USA 2006, 103, 11473-11478.
N. R. Rose, S. S. Ng, J. Mecinovic, B. M. Lienard, S. H. Bello, Z. Sun, M. A. McDonough, U. Oppermann and C. J. Schofield, J Med Chem, 2008, 51, 7053-7056.
Sakurai, M., Rose, N., Schultz, L., Quinn, A., Jadhav, A., Ng, S., Oppermann, U., Schofield, C.J. and Simeonov, A. Mol BioSystems 2009, in press.
S. Michael, D. Auld, C. Klumpp, A. Jadhav, W. Zheng, N. Thorne, C. Austin, J. Inglese and A. Simeonov, Assay Drug Dev Technol, 2008, 6, 637-657.
NIH Chemical Genomics Center [NCGC]
Structural Genomics Consortium [SGC]
Please see related BioAssays for all protocols relevant to this probe development project: AID 2147
Keywords: JMJD2, JMJD2E, Human 2-Oxoglutarate Oxygenase, qHTS
AID 2147: This assay is the primary screen of JMJD2E 226,938 samples.
This summary assay will be updated as additional screens are run and small molecule probes have been developed.
Categorized Comment - additional comments and annotations
From MLP Probe Report:
Probe count: 1
MLP Probe ML# for probe 1: ML324
PubChem Substance ID (SID) for probe 1: 85736407
PubChem Compound ID (CID) for probe 1: 44143209
Probe type for probe 1: Inhibitor
IC50/EC50 (nM) for probe 1: 920
Target for probe 1: JMJD2E (gi: 221046486)
Disease relevance for probe 1: Herpes Simplex
Anti-target for probe 1: LSD1
Fold selectivity for probe 1: n/a
NCBI Book chapter link for probe 1: http://www.ncbi.nlm.nih.gov/books/NBK169450/ (ID: 3060590)
Grant number for probe 1: MH084681
NCBI Book chapter title for probe 1: Discovery of ML324, a JMJD2 demethylase inhibitor with demonstrated antiviral activity