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BioAssay: AID 504391

qHTS Assay for Inhibitors of BAZ2B: Summary

BAZ2B (bromodomain adjacent to zinc finger domain, 2B) belongs to a family of ubiquitously expressed bromodomain containing proteins, which biological function has not yet been elucidated. However, it is suggested that BAZ2B has a similar function as the Drosophila Acf1 protein which regulates nucleosome mobilization through the ATP-dependent chromatin remodelling factor ISWI [1], resulting in alteration in the translational position of the histone and hence transcriptional regulation. The interaction of BAZ2B with ISWI mediated by the BAZ1 motif has recently been described [2]. ..more
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AID: 504391
Data Source: NCGC (BAZ2720)
BioAssay Type: Summary, Candidate Probes/Leads with Supporting Evidence
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-03-01
Target
Related Experiments
AIDNameTypeComment
504333qHTS Assay for Inhibitors of BAZ2BConfirmatorydepositor-specified cross reference: qHTS assay
588809qHTS Assay for Inhibitors of BAZ2B: Hit ValidationConfirmatorydepositor-specified cross reference
588859qHTS Assay for Inhibitors of BAZ2B: Hit Validation in AlphaScreen Counterscreen.Confirmatorydepositor-specified cross reference
Description:
BAZ2B (bromodomain adjacent to zinc finger domain, 2B) belongs to a family of ubiquitously expressed bromodomain containing proteins, which biological function has not yet been elucidated. However, it is suggested that BAZ2B has a similar function as the Drosophila Acf1 protein which regulates nucleosome mobilization through the ATP-dependent chromatin remodelling factor ISWI [1], resulting in alteration in the translational position of the histone and hence transcriptional regulation. The interaction of BAZ2B with ISWI mediated by the BAZ1 motif has recently been described [2].

There is a need for chemical probes to allow the elucidation of the roles of this protein in health and disease and hence a quantitative high-throughput screen [3,4] was developed. The protocol is based on the AlphaScreen (PerkinElmer) displacement assay developed by NCGC [5], but uses the peptide ligand Biot-H3K14Ac, a high affinity binding partner for BAZ2B, in an AlphaScreen format.

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 [6]. The top compound concentration tested was 110uM in this assay, which increases number of actives and potential artifacts. 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] Eberharter, A. et al. ACF1 improves the effectiveness of nucleosome mobilization by ISWI through PHD-histone contacts. The EMBO journal 23, 4029-4039. 2004. PMID: 15457208

[2] Jones, M.H. et al. A novel family of bromodomain genes. Genomics 63, 40-45. 2000. PMID: 10662543

[3] Yasgar, et al. Compound Management for Quantitative High-Throughput Screening. JALA. 2008 Apr;13(2):79-89. PMID: 18496600

[4] Inglese, et al. Quantitative high-throughput screening: a titration-based approach that efficiently identifies biological activities in large chemical libraries. Proc Natl Acad Sci. 1;103(31):11473-8. 2006. PMID: 16864780

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

[6] 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 linked AIDs for detailed protocols 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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