Reversible Regulation of Enzyme Activity by pH-Responsive Encapsulation in DNA Nanocages

Abstract

Reversible regulation of enzyme activity by chemical and physical stimuli is often achieved by incorporating stimuli-responsive domains in the enzyme of interest. However, this method is suitable for a limited number of enzymes with well-defined structural and conformational changes. In this study, we present a method to encapsulate enzymes in a DNA cage that could transform its conformation depending on the pH, allowing reversible control of the accessibility of the enzyme to the surrounding environment. This enabled us to regulate various properties of the enzyme, such as its resistance to protease-dependent degradation, binding affinity to the corresponding antibody, and most importantly, enzyme activity. Considering that the size and pH responsiveness of the DNA cage can be easily adjusted by the DNA length and sequence, our method provides a broad-impact platform for controlling enzyme functions without modifying the enzyme of interest.

In Silico and In Cell Analysis of Openable DNA Nanocages for miRNA Silencing

“…2020, 21, 61 2 of 11 transition from a "folded" to an "unfolded" form for the transport and release of triplex-specific binding molecules [13]. Tetrahedral DNA cages have been modified with the use of DNA oligonucleotides with pH-sensitive i-motif, to encapsulate an enzyme inside them [14]. DNA nanostructures have also been functionalized to selectively interact with intracellular miRNA, mainly to detect their concentration, using electrochemical current or fluorescence signals [15][16][17].…”

Biomimetic Compartments Scaffolded by Nucleic Acid Nanostructures

“…A light‐triggered release of bioactive cargoes of proteins and small molecules from a DNA nanocage was reported by Kohman et al, which was achieved through the incorporation of a photolabile crosslinker (Figure g) . Kim et al designed a pH‐switchable DNA tetrahedron for regulating protein stability against protease digestion, protein‐antibody binding, and enzyme activity (Figure h) . The Andersen group designed a DNA nanovault to control the access of substrate molecules to encapsulated enzymes by the reversible opening (accessible to substrate) and closing (inaccessible) of the cage (Figure i) …”

“…h) pH‐switched open and close of a DNA cage for regulating the stability and activity of encapsulated enzymes. Reproduced with permission . Copyright 2017, American Chemical Society.…”

DNA-Scaffolded Proximity Assembly and Confinement of Multienzyme Reactions

“…Kim et al designed a pH-switchable DNA tetrahedron for regulating protein stability against protease digestion, protein-antibody binding and enzyme activity (Fig. 8d) [113]. Andersen and coworkers designed a DNA nanovault to control the access of substrate molecules to encapsulated enzymes by the reversible opening (accessible to substrate) and closing (inaccessible) of the cage (Fig.…”

“…d A pH-switchable DNA tetrahedron for regulating protein stability and activity. Reproduced from Kim et al [113], with permission, copyright 2017, American Chemical Society. e A DNA nanovault with reversible opening and closing to regulate enzyme-substrate accessibility.…”