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BioAssay: AID 378

Compound Screen Assay, Human AASDHPPT

The apparent binding of compounds to Human AASDHPPT has been measured using differential scanning fluorimetry (DSF) technique. ..more
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 Tested Compounds
 Tested Compounds
All(4)
 
 
Active(4)
 
 
 Tested Substances
 Tested Substances
All(4)
 
 
Active(4)
 
 
 Related BioAssays
 Related BioAssays
AID: 378
Data Source: SGCOxCompounds (AASDHPPT Compound Screen)
BioAssay Type: Primary, Primary Screening, Single Concentration Activity Observed
BioAssay Version:
Deposit Date: 2006-04-21
Modify Date: 2006-10-24

Data Table ( Complete ):           Active    All
Target
BioActive Compounds: 4
Description:
The apparent binding of compounds to Human AASDHPPT has been measured using differential scanning fluorimetry (DSF) technique.

In this approach, the unfolding of the protein is monitored via a fluorescent hydrophobicity-sensing dye: SYPRO orange (Invitrogen). The hyperbolic curve of increase in fluorescence intensity versus temperature shows the unfolding process. The point of inflexion of this trace provides the measured parameter known as Tm. A difference in Tm between a control without the compound and an experiment with the compound present is called a TmShift. Positive shifts indicate that the protein is more stable in the presence of the compound, indicating that the compound is bound. Since it is not possible to quantitatively compare TmShift values from different experiments containing different components, we mark positive shifts of more than 3 degrees celcius as having a PubChem assay score of 100, and all other shifts with a score of 0.
AASDHPPT catalyzes the transfer of the 4'-phosphopantetheine moiety of coenzyme A to different carrier proteins, the most significant being the acyl carrier protein domains of cytosolic and mitochondrial fatty acid synthase enzymes. This modification of acyl-carrier proteins associated with type I and II fatty acids synthases is essential for lipid synthesis pathways.
Protocol
Dye concentration: 1:1000 (structure of dye is undisclosed)
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1expt1_Tm_ShiftChange in melting temperature relative to controlFloatCelcius
2expt1_Compound_ConcentrationConcentration of CompoundFloatμM
3expt1_Protein_ConcentrationConcentration of ProteinFloatμM
4expt1_Buffer_pH_SaltConditions: Buffer, pH, salt and additivesString
5expt1_MethodDSF MethodString
6expt2_Tm_ShiftChange in melting temperature relative to controlFloatCelcius
7expt2_Compound_ConcentrationConcentration of CompoundFloatμM
8expt2_Protein_ConcentrationConcentration of ProteinFloatμM
9expt2_Buffer_pH_SaltConditions: Buffer, pH, salt and additivesString
10expt2_MethodDSF MethodString
11expt3_Tm_ShiftChange in melting temperature relative to controlFloatCelcius
12expt3_Compound_ConcentrationConcentration of CompoundFloatμM
13expt3_Protein_ConcentrationConcentration of ProteinFloatμM
14expt3_Buffer_pH_SaltConditions: Buffer, pH, salt and additivesString
15expt3_MethodDSF MethodString
16expt4_Tm_ShiftChange in melting temperature relative to controlFloatCelcius
17expt4_Compound_ConcentrationConcentration of CompoundFloatμM
18expt4_Protein_ConcentrationConcentration of ProteinFloatμM
19expt4_Buffer_pH_SaltConditions: Buffer, pH, salt and additivesString
20expt4_MethodDSF MethodString

Data Table (Concise)
Classification
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