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BioAssay: AID 485341

qHTS Inhibitors of AmpC Beta-Lactamase (assay without detergent)

This aggregation profiling approach exploits the sensitivity of aggregate formation to detergent. Inhibition of b-lactamase is measured in the presence and absence of 0.01% Triton X-100. This particular assay had the prescence of 0.01% Triton X-100. See Pubchem assay "Promiscuous and Specific Inhibitors of AmpC Beta-Lactamase (assay without detergent)" for related screen. Compounds that inhibit only in the absence of detergent are considered likely promiscuous aggregators. ..more
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 Tested Compounds
 Tested Compounds
All(330683)
 
 
Active(1730)
 
 
Inactive(325710)
 
 
Inconclusive(3251)
 
 
 Tested Substances
 Tested Substances
All(331108)
 
 
Active(1730)
 
 
Inactive(326127)
 
 
Inconclusive(3251)
 
 
AID: 485341
Data Source: NCGC (AMPC203)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-10-04

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 1730
Related Experiments
AIDNameTypeComment
584Promiscuous and Specific Inhibitors of AmpC Beta-Lactamase (assay with detergent)Confirmatorydepositor-specified cross reference: Promiscuous and Specific Inhibitors of AmpC Beta-Lactamase (assay with detergent) - a screen of the
585Promiscuous and Specific Inhibitors of AmpC Beta-Lactamase (assay without detergent)Confirmatorydepositor-specified cross reference: Promiscuous and Specific Inhibitors of AmpC Beta-Lactamase (assay without detergent) - a screen old
1002Confirmation Concentration-Response Assay for Inhibitors of AmpC Beta-Lactamase (assay with detergent)Confirmatorydepositor-specified cross reference: Confirmation Concentration-Response Assay for Inhibitors of AmpC Beta-Lactamase (assay with detergen
1003Confirmation Cuvette-Based Assay for Inhibitors of AmpC Beta-Lactamase (assay with detergent)Confirmatorydepositor-specified cross reference: Confirmation Cuvette-Based Assay for Inhibitors of AmpC Beta-Lactamase (assay with detergent)
485291Probe Development Summary for Inhibitors of AmpC Beta LactamaseSummarydepositor-specified cross reference: Probe Development Summary for Inhibitors of AmpC Beta Lactamase
485294qHTS Inhibitors of AmpC Beta-Lactamase (assay with detergent)Confirmatorydepositor-specified cross reference: qHTS Inhibitors of AmpC Beta-Lactamase (assay with detergent)
Description:
NIH Molecular Libraries Probe Production Network [MLPCN]
NIH Chemical Genomics Center [NCGC]
Assay Provider: Professor Brian Shoichet, UCSF

Assay Overview:
This aggregation profiling approach exploits the sensitivity of aggregate formation to detergent. Inhibition of b-lactamase is measured in the presence and absence of 0.01% Triton X-100. This particular assay had the prescence of 0.01% Triton X-100. See Pubchem assay "Promiscuous and Specific Inhibitors of AmpC Beta-Lactamase (assay without detergent)" for related screen. Compounds that inhibit only in the absence of detergent are considered likely promiscuous aggregators.
A screen of 1,030 compounds was performed in Brian Shoichet's lab in 96-well format [1]. AmpC activity was measured by following the absorbance at 480 nm. Nitrocefin is a chromogenic substrate: while nitrocefin is yellow (lmax 390 nm), the product of its hydrolysis is red (lmax 486 nm, 17,000 M-1). AmpC beta lactamase was provided by Shoichet Lab, UCSF.
Protocol
Buffer:
20 mM phosphate buffer, pH 7.0
Triton X-100: No Detergent screen: 0.0001%, Detergent screen: 0.01%.
Reagents/Controls:
Buffer in columns 3 and 4 as negative control (no enzyme).
5.3 nM AmpC in columns 1, 2, 5-48. Titration of UCSF compound 9 control (from 10 mM, then 1:2) pin-transferred to column 2. Column 1 is neutral (100% activity).
Assay Steps:
Six uL of reagents were dispensed to 1536-well Greiner black clear bottom plates. Compounds and controls (23 nL) were transferred via Kalypsys PinTool. The plates were incubated for 15 min at room temperature, and then 2 uL of 800 uM solution of nitrocefin was added to start the reaction. The plates were immediately transferred to and read every 20 seconds for 4 min on ViewLux (Perkin-Elmer) High-throughput CCD imager using 480 nm absorbance protocol. During dispense, reagent bottles were kept submerged into 4 deg C water bath.
Keywords: NIH Roadmap, MLSCN, MLI, MLSMR, qHTS, NCGC, AmpC Beta-Lactamase, Aggregation
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
5Curve_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
6Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
7Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
8Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
9Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
10Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
11Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
12Excluded_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
13Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
14Activity at 0.018 uM (0.0183μM**)% Activity at given concentration.Float%
15Activity at 0.091 uM (0.0914367μM**)% Activity at given concentration.Float%
16Activity at 0.457 uM (0.457μM**)% Activity at given concentration.Float%
17Activity at 2.290 uM (2.29μM**)% Activity at given concentration.Float%
18Activity at 11.40 uM (11.4μM**)% Activity at given concentration.Float%
19Activity at 57.10 uM (57.1μM**)% Activity at given concentration.Float%
20Activity at 114.0 uM (114μM**)% Activity at given concentration.Float%
21Compound QCSource of compound QCString

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH079825-01

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
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