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BioAssay: AID 269135

Inhibition of Escherichia coli KPR

Mapping interactions at protein-ligand binding sites is an important aspect of understanding many biological reactions and a key part of drug design. In this paper, we have used a fragment-based approach to probe "hot spots" at the cofactor-binding site of a model dehydrogenase, Escherichia coli ketopantoate reductase. Our strategy involved the breaking down of NADPH (Kd = 300 nM) into more ..
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 Tested Compounds
 Tested Compounds
All(7)
 
 
Unspecified(7)
 
 
 Tested Substances
 Tested Substances
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Unspecified(7)
 
 
 Related BioAssays
 Related BioAssays
AID: 269135
Data Source: ChEMBL (382750)
Depositor Category: Literature, Extracted
BioAssay Version:
Deposit Date: 2010-05-25
Modify Date: 2013-11-18

Data Table ( Complete ):           All
Tested Compounds:
Description:
Title: Probing hot spots at protein-ligand binding sites: a fragment-based approach using biophysical methods.

Abstract: Mapping interactions at protein-ligand binding sites is an important aspect of understanding many biological reactions and a key part of drug design. In this paper, we have used a fragment-based approach to probe "hot spots" at the cofactor-binding site of a model dehydrogenase, Escherichia coli ketopantoate reductase. Our strategy involved the breaking down of NADPH (Kd = 300 nM) into smaller fragments and the biophysical characterization of their binding using WaterLOGSY NMR spectroscopy, isothermal titration calorimetry (ITC), and inhibition studies. The weak binding affinities of fragments were measured by direct ITC titrations under low c value conditions. The 2'-phosphate and the reduced nicotinamide groups were found to contribute a large part of the binding energy. A combination of ITC and site-directed mutagenesis enabled us to locate the fragments at separate hot spots on opposite ends of the cofactor-binding site. This study has identified structural determinants for cofactor recognition that represent a blueprint for future inhibitor design.
(PMID: 16884311)
Comment
Putative Target:

ChEMBL Target ID: 100777
Target Type: SINGLE PROTEIN
Pref Name: Ketopantoate reductase
Synonyms: 2-dehydropantoate 2-reductase;Ketopantoate reductase;KPA reductase;KPR;
Gene Name: apbA;JW0415;panE;
Protein Accession: P0A9J4;
Protein GI: 71159619;
Organism: Escherichia coli K-12
Tax ID: 83333
Target Classification: enzyme
Confidence: Homologous single protein target assigned
Relationship Type: Homologous protein target assigned
Categorized Comment
ChEMBL Assay Type: Binding

ChEMBL Assay Data Source: Scientific Literature

Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1Ki*Ki PubChem standard valueFloatμM
2Ki activity commentKi activity commentString
3Ki standard flagKi standard flagInteger
4Ki qualifierKi qualifierString
5Ki published valueKi published valueFloatmM
6Ki standard valueKi standard valueFloatnM
7Ki data validityKi data validityString
8Km activity commentKm activity commentString
9Km standard flagKm standard flagInteger
10Km qualifierKm qualifierString
11Km published valueKm published valueFloatmM
12Km standard valueKm standard valueFloatnM
13Km data validityKm data validityString

* Activity Concentration.

Data Table (Concise)
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