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BioAssay: AID 488978

High-Throughput Screening for Modulators of Cytosolic Chaperonin Activity: MmCpn Primary Screen

Chaperonins are key components of the cellular chaperone machinery. These large complexes consist of two stacked 7-9 membered rings, each containing a central cavity. Substrates are folded within the cavity in an ATP-dependent manner. TRiC is a complex of eight different, essential proteins. As each of the genes encoding the eight subunits is essential for cell viability and no protocol has more ..
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 Tested Compounds
 Tested Compounds
All(1266)
 
 
Active(1)
 
 
Inactive(1250)
 
 
Inconclusive(15)
 
 
 Tested Substances
 Tested Substances
All(1280)
 
 
Active(1)
 
 
Inactive(1264)
 
 
Inconclusive(15)
 
 
AID: 488978
Data Source: NCGC (CHAP881)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2010-11-02

Data Table ( Complete ):           Active    All
Target
BioActive Compound: 1
Depositor Specified Assays
AIDNameTypeComment
488991High-Throughput Screening for Modulators of Cytosolic Chaperonin Activity: Summarysummary
Description:
Chaperonins are key components of the cellular chaperone machinery. These large complexes consist of two stacked 7-9 membered rings, each containing a central cavity. Substrates are folded within the cavity in an ATP-dependent manner. TRiC is a complex of eight different, essential proteins. As each of the genes encoding the eight subunits is essential for cell viability and no protocol has been developed to overexpress this enzyme to high levels, several groups have turned to the archaeal homolog of TRiC from Methanococcus maripaludis (MmCpn) as a model system for the study of the type II chaperonin family.
Protocol
MmCpn was kindly provided by the laboratory of Judith Frydman.

1. Dispense 1.5 ul enzyme of either TriC at 100 nM or MmCpn at 50 nM final concentration.
2. Transfer 23 nl of compound to plate, incubate 5 min at RT.
3. 1.5 ul ATP at 30 uM at final concentration dispensed into plate.
4. Incubate for 10 minutes at 37C (MmCPN), 30C (TriC).
5. 3 ul of the Cisbio ADP Transcreener HTRF detection reagent prepared according to protocol dispensed into plate.
6. Incubate for 5 minutes.
7. Read using EnVision plate reader at Excitation = 320nm and Emission = 590nm, 665nm
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 0.00492 uM (0.00492μM**)% Activity at given concentration.Float%
15Activity at 0.025 uM (0.0246μM**)% Activity at given concentration.Float%
16Activity at 0.123 uM (0.123μM**)% Activity at given concentration.Float%
17Activity at 0.615 uM (0.615μM**)% Activity at given concentration.Float%
18Activity at 3.080 uM (3.08μM**)% Activity at given concentration.Float%
19Activity at 15.40 uM (15.4μM**)% Activity at given concentration.Float%
20Activity at 76.90 uM (76.9μM**)% Activity at given concentration.Float%
21Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: DA030554-01A1

Data Table (Concise)
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