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BioAssay: AID 504398

qHTS Assay for Inhibitors of GCN5L2: Summary

Histone acetyltransferase (HAT) enzymes transfer a methyl group from acetyl-coenzyme A to the epsilon-amino group of conserved lysine residues of N-terminal histone tails. Acetylation results in charge neutralization of this positively charged amino acid, decreasing interactions with the positively charged DNA backbone and making the DNA more accessible for transcription. HATs have been more ..
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AID: 504398
Data Source: NCGC (GCN500)
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
504327qHTS Assay for Inhibitors of GCN5L2Confirmatorydepositor-specified cross reference: qHTS
588347qHTS Assay for Inhibitors of GCN5L2: Hit confirmationConfirmatorydepositor-specified cross reference
Description:
Histone acetyltransferase (HAT) enzymes transfer a methyl group from acetyl-coenzyme A to the epsilon-amino group of conserved lysine residues of N-terminal histone tails. Acetylation results in charge neutralization of this positively charged amino acid, decreasing interactions with the positively charged DNA backbone and making the DNA more accessible for transcription. HATs have been recognized as an emerging drug target for treatment of cancer, diabetes, asthma, chronic obstructive pulmonary disease, cardiac disease, and viral infection [1]. Protein identified with HAT activity include the 5 yeast homolog like 2 (GCN5L2, a.ka. Histone acetyltransferase KAT2A) which acetylates histone H3 at Lys9, 14, 18, 13, and to lesser extent histones H4 and H2B. This activates transcription of the gene by modulating chromatin structure and installing the acetyllysine recognition mark for bromodomain-containing coactivator proteins.
A quantitative high throughput screen [2,3] was developed to identify small molecule inhibitors. The assay is based on the principle that it measures formation of reduced CoA with the pro-fluorescent maleimide derivative ThioGlo1 that becomes fluorescent upon reaction with the free thiol of CoA.

A large number of artifacts are expected in this primary screen, compounded by the assay format (fluorescent artifacts) and by the high top concentration tested for each substance. This BioAssay data deposition should be used with caution, and counterscreen and secondary assay validation is essential since very few actives from the primary screen are expected to be true inhibitors of GCN5L2.

[1] Esteller, M. Epigenetics in Cancer. N Engl J Med 358(11), 1148-1159. (2008). PMID: 18337604

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

[3] 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

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 assay AIDs for a detailed description of 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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