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BioAssay: AID 397449

Activation of human Go protein alpha subunit expressed in Escherichia coli assessed as protein bound [35S]GTPgammaS levels at 50 uM by scintillation spectroscopy

This paper documents a serious problem met during the testing of Gi protein-activating properties of a new series of synthetic compounds by measuring the induced binding of [(35)S]GTPgammaS to different subtypes of Gi protein. The problem arose from the strong affinity between [(35)S]GTPgammaS and the tested compounds, that are characterized by several (2-4) positive charges and high lipophilicity. Apparently, such affinity yields insoluble, labelled complexes that, also in the absence of Gi protein, are retained on the filters and give rise to false positive results. ..more
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 Tested Compounds
 Tested Compounds
All(1)
 
 
Unspecified(1)
 
 
 Tested Substances
 Tested Substances
All(1)
 
 
Unspecified(1)
 
 
 Related BioAssays
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AID: 397449
Data Source: ChEMBL (545478)
Depositor Category: Literature, Extracted
BioAssay Version:
Deposit Date: 2010-05-26
Modify Date: 2014-08-23

Data Table ( Complete ):           View All Data
Target
Sequence: RecName: Full=Guanine nucleotide-binding protein G(o) subunit alpha
Description ..   
Protein Family: Alpha subunit of G proteins (guanine nucleotide binding)
Comment ..   

Gene:GNAO1     Related Protein 3D Structures     More BioActivity Data..
Tested Compound:
Description:
Title: [35S]GTP gamma S binding studies of amphiphilic drugs-activated Gi proteins: a caveat.

Abstract: This paper documents a serious problem met during the testing of Gi protein-activating properties of a new series of synthetic compounds by measuring the induced binding of [(35)S]GTPgammaS to different subtypes of Gi protein. The problem arose from the strong affinity between [(35)S]GTPgammaS and the tested compounds, that are characterized by several (2-4) positive charges and high lipophilicity. Apparently, such affinity yields insoluble, labelled complexes that, also in the absence of Gi protein, are retained on the filters and give rise to false positive results.
(PMID: 19289280)
Categorized Comment - additional comments and annotations
From BioAssay Depositor:
Assay Type: Binding
Target Type: Target is a single protein chain
Assay Data Source: Scientific Literature
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1Activity activity commentActivity activity commentString
2Activity standard flagActivity standard flagInteger
3Activity qualifierActivity qualifierString
4Activity published valueActivity published valueFloatfmol
5Activity standard valueActivity standard valueFloatfmol

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