Supramolecular Tandem Enzyme Assays

We conceptualize a novel approach towards enzyme assays based on the reversible and competitive binding of a fluorescent dye and the substrate as well as product of an enzymatic reaction to a macrocyclic host. This method was termed “supramolecular tandem assay”, and has been applied to inhibitor an...

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Bibliographic Details
Published in:Chemistry : a European journal 2012-03, Vol.18 (12), p.3444-3459
Main Authors: Dsouza, Roy N., Hennig, Andreas, Nau, Werner M.
Format: Article
Language:English
Subjects:
Amino Acids - chemistry
Assaying
Binding, Competitive
Catalysts
Chemistry
Enzyme Assays
Enzymes
Fluorescence
fluorescent dyes
Fluorescent Dyes - chemistry
Inhibitors
macrocycles
Macrocyclic Compounds - chemistry
molecular recognition
Monitoring
Screening
Sensor arrays
Stereoisomerism
Substrate Specificity
supramolecular chemistry
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Summary:We conceptualize a novel approach towards enzyme assays based on the reversible and competitive binding of a fluorescent dye and the substrate as well as product of an enzymatic reaction to a macrocyclic host. This method was termed “supramolecular tandem assay”, and has been applied to inhibitor and activator screening, sensor array development, and enantiomeric excess determination of amino acids. The simple and rapid read‐out by fluorescence allows their straightforward implementation into high‐throughput screening. The combination of macrocycles with fluorescent dyes affords supramolecular reporter pairs, which can be employed for the monitoring of catalytic, particularly biocatalytic, reactions. The resulting assays are unselective time‐resolved variants of indicator displacement and uptake sensing systems. They are applicable to a large range of analytes and allow direct real‐time monitoring in homogeneous solution, with the possibility of performing inhibitor and catalyst screening.
ISSN:0947-6539
1521-3765
DOI:10.1002/chem.201103364