Abstract
Methods for the detection of specific interactions between diverse proteins and various small-molecule ligands are of significant importance in understanding the mechanisms of many critical physiological processes of organisms. The techniques also represent a major avenue to drug screening, molecular diagnostics, and public safety monitoring. Terminal protection assay of small molecule-linked DNA is a demonstrated novel methodology which has exhibited great potential for the development of simple, sensitive, specific and high-throughput methods for the detection of small molecule–protein interactions. Herein, we review the basic principle of terminal protection assay, the development of associated methods, and the signal amplification strategies adopted for performance improving in small molecule–protein interaction assay.
Highlights
The affinity binding of small molecules with their target proteins relies on noncovalent but specific interactions, and small molecules that interact with proteins in this way serve as the affinity ligands of the associated proteins [1,2]
The review traces the recent development in the field of small molecule–protein interaction assays upon the terminal protection of small molecule-labeled DNA
Terminal protection is a generalized discovery demonstrated by Jiang et al [17], that small molecule–DNA chimeras could be protected from degradation by various DNA exonucleases
Summary
The affinity binding of small molecules with their target proteins relies on noncovalent but specific interactions, and small molecules that interact with proteins in this way serve as the affinity ligands of the associated proteins [1,2]. Some classic techniques, including affinity chromatography [9,10], kinetic capillary electrophoresis [11,12], fluorescence polarization [13,14] and surface plasmon resonance [15,16], have been developed for the detection of small molecule–protein interactions Problems such as the complex fixation of proteins or small molecules, limited sensitivity, potential nonspecific adsorption, or the requirement of sophisticated instruments frequently limit their widespread application. Jiang and colleagues proposed a completely different concept of terminal protection assay for the investigation of small molecule–protein interactions [17] They found an interesting phenomenon: the binding of a protein to a small molecule moiety at one terminus of a DNA module could protect the DNA from digestion by nucleases. Some methods that share the concept of DNA protection assay are discussed
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