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BioAssay: AID 652037

qHTS Assay for Inhibitors of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1): Insulin MOA Assay

The selenoprotein thioredoxin reductase (TrxR; EC 1.8.1.9) is a FAD containing homodimeric pyridine nucleotide-disulfide oxidoreductase with many cellular roles. Together with NADPH and its prime substrate thioredoxin (Trx), the enzyme forms the core of the Trx system. The mammalian Trx system exerts a wide spectrum of functions including redox regulation, antioxidant defense, regulation of more ..
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 Tested Compounds
 Tested Compounds
All(54)
 
 
Active(10)
 
 
Inactive(44)
 
 
 Tested Substances
 Tested Substances
All(54)
 
 
Active(10)
 
 
Inactive(44)
 
 
AID: 652037
Data Source: NCGC (TrxR203)
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2013-02-12

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 10
Related Experiments
AIDNameTypeComment
488771Probe Development Summary for Modulators of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1)Summarydepositor-specified cross reference
488772qHTS Assay for Substrates of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1)Confirmatorysame project related to Summary assay
488773qHTS Assay for Inhibitors of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1)Confirmatorysame project related to Summary assay
588453qHTS Assay for Inhibitors of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1): qHTSConfirmatorysame project related to Summary assay
588456qHTS Assay for Substrates of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1): qHTSConfirmatorysame project related to Summary assay
652020qHTS Assay for Inhibitors of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1): Counterassay in Glutathione Reductase (GR)Confirmatorysame project related to Summary assay
652023qHTS Assay for Inhibitors of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1): Hit Validation in Primary ScreenConfirmatorysame project related to Summary assay
652256qHTS Assay for Inhibitors of Mammalian Selenoprotein Thioredoxin Reductase 1 (TrxR1): DTNB AssayOthersame project related to Summary assay
Description:
The selenoprotein thioredoxin reductase (TrxR; EC 1.8.1.9) is a FAD containing homodimeric pyridine nucleotide-disulfide oxidoreductase with many cellular roles. Together with NADPH and its prime substrate thioredoxin (Trx), the enzyme forms the core of the Trx system. The mammalian Trx system exerts a wide spectrum of functions including redox regulation, antioxidant defense, regulation of transcription factors as well as support of cell growth and replication. Many of these functions involve the reduction of Trx, which may subsequently reduce a number of different substrates including ribonucleotide reductase, peroxiredoxins or NFkB. Mammalian TrxR itself also has a broad substrate specificity, reducing both protein and non-protein substrates, including low molecular weight compounds such as dehydroascorbate, lipoic acid, ubiquinone, and juglone. In addition, several drugs in clinical use for anticancer treatment are indeed known to target TrxR1.

This MOA Insulin assay seeks to further characterize the cherry picks.

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

MLPCN Grant: MH090846
Assay Provider: Elias Arner, Karolinska Institute
Protocol
0.8 microM TrxR1 is reduced by 200 microM NADPH in TE buffer [50 mM Tris-HCl (pH 7.5), 2 mM EDTA] and incubated at room temperature with increasing concentrations compound for various time points. Duplicate aliquots are removed and added to a master mix containing 20 microM Trx1, 160 microM insulin and 200 microM NADPH in TE buffer. NADPH consumption at 30 degrees C is followed at 340 nm using a VersaMax microplate reader. The natural logarithm of the percentage of inactivation compared to DMSO treated control is then plotted against time. The resulting slopes, the apparent first-order inactivation rate constants (kobs), are plotted against the inhibitor concentrations resulting in the second-order rate constant kinact.
Comment
Compounds were an IC50 was determined < 100 uM were considered "active" and assigned a score of 90; compounds were the IC50 >= 100 uM or could not be determined were considered "inactive" and assigned a score of 10.
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Insulin IC50 uMIntegerμM
Additional Information
Grant Number: MH090846

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