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BioAssay: AID 540353

qHTS for Inhibitos of Bacillus subtilis Sfp phosphopantetheinyl transferase (PPTase): Chemistry Optimization Followup in B. Subtilis HM489

Phosphopantetheinyl transfer is a process essential to cellular viability and maintenance. Cellular permeable and stable compounds acting as PPTase inhibitors are anticipated to be antimicrobial agents in organisms with complimenting genotypes. The primary test organism in these experiments, B. subtilis, possesses two PPTase loci in the genome, acpS and sfp. This experiment evaluated a series of more ..
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 Tested Compounds
 Tested Compounds
All(204)
 
 
Active(92)
 
 
Inactive(56)
 
 
Inconclusive(59)
 
 
 Tested Substances
 Tested Substances
All(219)
 
 
Active(99)
 
 
Inactive(58)
 
 
Inconclusive(62)
 
 
AID: 540353
Data Source: NCGC (PPT804)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2011-08-15
Hold-until Date: 2012-08-14
Modify Date: 2012-08-14

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 92
Depositor Specified Assays
AIDNameTypeComment
1819Probe Development Summary of Inhibitors of Bacillus subtilis Sfp phosphopantetheinyl transferase (PPTase)summarySummary AID
Description:
Phosphopantetheinyl transfer is a process essential to cellular viability and maintenance. Cellular permeable and stable compounds acting as PPTase inhibitors are anticipated to be antimicrobial agents in organisms with complimenting genotypes. The primary test organism in these experiments, B. subtilis, possesses two PPTase loci in the genome, acpS and sfp. This experiment evaluated a series of test compounds for antimicrobial activity with B. subtilis HM489 (genotype: acpS- and sfp+), a strain that contains a targeted lesion in the acpS locus and thus requires a functional Sfp gene product for viability. [Mootz HD, 2001] This strain is expected to be sensitized to cell-permeable Sfp inhibitors.

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

MLPCN Grant: MH083266-01
Assay Submitter (PI): Michael Burkart, University of California, San Diego
Protocol
Innoculum preparation. Bacillus subtilis HM489 is maintained on lysogeny broth (LB) solidified by the addition of 1.5% w/v agar. Single colonies were used to LB medium (2 mL) and shaken overnight at 30 masculineC. In the morning, this culture (100 uL) was used to seed a fresh LB medium (10 mL) and was shaken at 30 masculineC until the culture OD600 reached 0.5. This culture was diluted 1:100 in fresh LB medium to provide the inoculum below.

Bacterial susceptibility. LB medium (2 uL) is dispensed into wells of a sterile white 1536-well plate. Test compounds (23 nL) prepared as serial dilutions in DMSO are added to the plate by pintool transfer. Innoculum (2 uL) is added; the plates are covered with a vented Kalypsys assay lid and incubated at 30 masculineC. After 5 hrs, Bac-Titer Glo (4 uL; Promega Corp, Madison, WI) is added to the plates. They are incubated 10 minutes at room temperature and then the luminescence detected in a ViewLux multimodal plate reader.
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 0.0006451429 uM (0.000645143μM**)% Activity at given concentration.Float%
15Activity at 0.00194 uM (0.00193543μM**)% Activity at given concentration.Float%
16Activity at 0.00581 uM (0.00580629μM**)% Activity at given concentration.Float%
17Activity at 0.017 uM (0.017419μM**)% Activity at given concentration.Float%
18Activity at 0.052 uM (0.0522568μM**)% Activity at given concentration.Float%
19Activity at 0.157 uM (0.156771μM**)% Activity at given concentration.Float%
20Activity at 0.470 uM (0.470312μM**)% Activity at given concentration.Float%
21Activity at 1.411 uM (1.41093μM**)% Activity at given concentration.Float%
22Activity at 4.233 uM (4.2328μM**)% Activity at given concentration.Float%
23Activity at 12.70 uM (12.6984μM**)% Activity at given concentration.Float%
24Activity at 38.10 uM (38.0952μM**)% Activity at given concentration.Float%
25Activity at 114.3 uM (114.286μM**)% Activity at given concentration.Float%
26Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: MH083266

Data Table (Concise)
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