Articles
A CONVENIENT METHOD FOR THE SCREENING OF COMPOUNDS THAT INHIBIT SPECIFIC MOLECULAR INTERACTIONS USING THE ALKALINE PHOSPHATASE FUSION SYSTEM
Article number
678_6
Pages
51 – 57
Language
English
Abstract
The identification of molecules that can interact specifically with drug targets is an important step in the drug discovery process.
Here, we describe a convenient and versatile method for the screening of compounds from plant products that can inhibit molecular interactions of interest.
In this assay, enzymes, protein, or peptide ligands are fused to the amino terminus of bacterial alkaline phosphatase (BAP) using genetic engineering.
These alkaline phosphatase fusion proteins are then used as one-step probes for detecting specific molecular interactions with various targets that are immobilized on the wells of a microtiter plate or membrane.
Molecular interactions can be determined by colorimetric assay of alkaline phosphatase activity, which is highly specific and sensitive.
Identification of compounds that can interfere with specific molecular interaction is determined from the reduction of signal from colorimetric analysis.
Assays for different compounds at various conditions can be done at the same time.
The cost for the assay is low and the technique is relatively easy.
The identification of compounds that can antagonize specific molecular interactions should prove useful in the study of the nature of these interactions within the cell, as well as suggest leads for drug development.
In addition, this assay can be applied to determine the mechanism of actions or to predict possible adverse reactions of plant-derived compounds.
Here, we describe a convenient and versatile method for the screening of compounds from plant products that can inhibit molecular interactions of interest.
In this assay, enzymes, protein, or peptide ligands are fused to the amino terminus of bacterial alkaline phosphatase (BAP) using genetic engineering.
These alkaline phosphatase fusion proteins are then used as one-step probes for detecting specific molecular interactions with various targets that are immobilized on the wells of a microtiter plate or membrane.
Molecular interactions can be determined by colorimetric assay of alkaline phosphatase activity, which is highly specific and sensitive.
Identification of compounds that can interfere with specific molecular interaction is determined from the reduction of signal from colorimetric analysis.
Assays for different compounds at various conditions can be done at the same time.
The cost for the assay is low and the technique is relatively easy.
The identification of compounds that can antagonize specific molecular interactions should prove useful in the study of the nature of these interactions within the cell, as well as suggest leads for drug development.
In addition, this assay can be applied to determine the mechanism of actions or to predict possible adverse reactions of plant-derived compounds.
Authors
M. Yamabhai
Keywords
drug screening, molecular interaction, alkaline phosphatase, plant extracts
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