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BioAssay: AID 2421

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 ..
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AID: 2421
Data Source: NCGC (JMJD100)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-02-24
Target
Depositor Specified Assays
AIDNameTypeComment
2147qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)confirmatoryqHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)
2677Confirmation qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)confirmatory
2680Counterscreen for JMJD2E Inhibitors: qHTS Assay for Inhibitors of Formaldehyde Dehydrogenase (FDH)confirmatory
2687AlphaScreen Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)confirmatory
2688MALDI-TOF-MS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E)confirmatory
687009qHTS Assay for Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E): Counterscreen with LSD1confirmatory
493212Inhibitors of Human Jumonji Domain Containing 2E (JMJD2E): 8HQs - Round 1confirmatory
Description:
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.

References:
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]
Protocol
Please see related BioAssays for all protocols relevant to this probe development project: AID 2147
Comment
Keywords: JMJD2, JMJD2E, Human 2-Oxoglutarate Oxygenase, qHTS

Related BioAssays:

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.
Additional Information
Grant Number: U54 CDP

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