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BioAssay: AID 504404

qHTS Assay for Inhibitors of Histone Lysine Methyltransferase G9a: Summary

Methylated lysines, on the N-terminal tails of histone proteins serve as epigenetic markers to recruit factors that can then modify local chromatin structure to lead to functional consequences. G9a and other members of the SUV39 family of the SET domain-containing superfamily of histone lysine methyltransferases (HMT) have been identified to specifically methylate Lys9 of histone H3 (H3K9). G9a more ..
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AID: 504404
Data Source: NCGC (G9A500)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-03-02
Target
Related Experiments
AIDNameTypeComment
504332qHTS Assay for Inhibitors of Histone Lysine Methyltransferase G9aConfirmatorydepositor-specified cross reference: qHTS
588344qHTS Assay for Inhibitors of Histone Lysine Methyltransferase G9a: Hit ConfirmationConfirmatorydepositor-specified cross reference
588346qHTS Assay for Inhibitors of Histone Lysine Methyltransferase G9a: Hit Confirmation in b-rIgG CounterscreenConfirmatorydepositor-specified cross reference
Description:
Methylated lysines, on the N-terminal tails of histone proteins serve as epigenetic markers to recruit factors that can then modify local chromatin structure to lead to functional consequences. G9a and other members of the SUV39 family of the SET domain-containing superfamily of histone lysine methyltransferases (HMT) have been identified to specifically methylate Lys9 of histone H3 (H3K9). G9a catalyzes the mono- and di-methylation of H3K9 in mammalian euchromatic regions, where the resulting H3K9me2 is indicative of transcriptional repression. Hence, G9a has been recognized as a potential drug target for several human diseases, including cancer. The inhibition of G9a will result in transcriptional activation and work synergistically with DNA methyltransferase and histone deacetylase inhibitors, to kill cancer cells.

This qHTS assay for identification of G9a inhibitors is a chemiluminescence based AlphaScreen (PerkinElmer) [1]. Methylation of biotinylated-histone peptide is measured through specific antibody-based detection, in conjunction with streptavidin-coated donor and anti-IgG antibody-coated acceptor beads. The method is particularly well suited for detection of inhibitors acting by the desired histone peptide competitive mechanism and is applicable to testing other HMTs.

Although AlphaScreen has significant advantages with its utility in a variety of epigenetic target assays, the primary screening data provided in this deposition should be used with caution due to the prevalence of screening artifacts [2]. An AlphaScreen counterscreen is highly recommended to be run against any putative actives to eliminate non specific artifacts. Alternatively, one can use other AlphaScreen assays in PubChem to filter out promiscuous hits.

[1] Quinn et al. A chemiluminescence-based method for identification of histone lysine methyltransferase inhibitors. Mol Biosyst 6(5): 782-8. 2010. PMID: 20567762

[2] Baell,Holloway. New substructure filters for removal of pan assay interference compounds (PAINS) from screening libraries and for their exclusion in bioassays. J Med Chem. 53(7):2719-40. 2010. PMID: 20131845

NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
Structural Genomics Consortium (SGC)
NIH Grant: 5U54 MH084681-02
Protocol
Please see the linked assay AIDs for a detailed protocol for each assay.
Comment
This project is on-going and will be updated with findings at a later point.
Additional Information
Grant Number: MH084681-02

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