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BioAssay: AID 651804

qHTS for Small Molecule Inhibitors of the ERG Ets/DNA interaction: HTRF assay against cherry picks

The Ets family of proteins is a family of ~30 conserved transcription factors defined by the presence of an ~85 amino acid domain referred to as the Ets domain, which mediates sequence-specific DNA binding to a core DNA element GGAA/T.The Ets family member ERG has been linked to several neoplasms. ERG has been shown to be frequently over-expressed in prostate cancer. Perhaps more strikingly, ERG more ..
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 Tested Compounds
 Tested Compounds
All(1236)
 
 
Active(20)
 
 
Inactive(930)
 
 
Inconclusive(286)
 
 
 Tested Substances
 Tested Substances
All(1236)
 
 
Active(20)
 
 
Inactive(930)
 
 
Inconclusive(286)
 
 
AID: 651804
Data Source: NCGC (ERG201)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-11-21

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 20
Depositor Specified Assays
AIDNameTypeComment
624266qHTS for Small Molecule Inhibitors of the ERG Ets/DNA interaction: Summary.summarySummary AID
Description:
The Ets family of proteins is a family of ~30 conserved transcription factors defined by the presence of an ~85 amino acid domain referred to as the Ets domain, which mediates sequence-specific DNA binding to a core DNA element GGAA/T.The Ets family member ERG has been linked to several neoplasms. ERG has been shown to be frequently over-expressed in prostate cancer. Perhaps more strikingly, ERG and the Ets protein ETV1 have recently been shown to be the targets of chromosomal translocations with TMPRSS2 which are observed in 80% of prostate cancer patient samples. More extensive studies of the effects of over-expression of ERG in prostate cells have recently been carried out and have shown that over-expression of ERG results in increased invasion.

The importance of DNA-binding to the function of these proteins strongly suggests that inhibiting the DNA binding activity of ERG may be a mechanism to treat prostate cancer as well as other cancers where the activity of ERG plays a key role. We will carry out a high throughout screen to identify inhibitors of the interaction between ERG and DNA using a fluorescence polarization (FP) assay with a His-tagged ERG Ets domain and fluorescently-labeled DNA probes to develop small molecule inhibitors of this interaction as probes to test this hypothesis and lay the groundwork for the development of targeted therapies against ERG. This is the counterscreen against the cherry picks.

NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]

Grant: NS066464
PI Name: John Bushweller, University of Virgina
Protocol
2 ul of protein solution (200 nM ERG domain in 300mM KCl, 50mM Tris-HCl, 1mM EDTA, 1mM DTT, pH 7.5); 20 nl Control plate and compound library plate; 2 ul of 200 nM biotin-DNA; incubate for 10 min at RT; 1 ul of anti-His antibody conjugated to the long-life fluorescent cryptate donner deuterium (5ng/ul) and streptavidin conjugated to the acceptor compound Terbium (0.2 ng/ul) mix; incubate 10 min and measure FRET (Excitation = 320 nm, Emission = 615, 665 nm).
Comment
Compound Ranking:

1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Activity_ScoreActivity score.Integer
6Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.00492 uM (0.00492μM**)% Activity at given concentration.Float%
16Activity at 0.025 uM (0.0246μM**)% Activity at given concentration.Float%
17Activity at 0.123 uM (0.123μM**)% Activity at given concentration.Float%
18Activity at 0.615 uM (0.615μM**)% Activity at given concentration.Float%
19Activity at 3.080 uM (3.08μM**)% Activity at given concentration.Float%
20Activity at 15.40 uM (15.4μM**)% Activity at given concentration.Float%
21Activity at 76.90 uM (76.9μM**)% Activity at given concentration.Float%
22Activity at 154.0 uM (154μM**)% Activity at given concentration.Float%
23Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: NS066464

Data Table (Concise)
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