We describe the operational principle, synthesis, and applications of the enzyme-DNA chimeras. These are supramolecular constructions where a DNA spring is coupled to an enzyme and introduces artificial allosteric control of the enzyme. This method is universal and can be applied to various enzymes and proteins. In addition, this method is versatile as the stresses applied by the DNA spring on the enzymes can be fine-tuned semi-continuously and thus their enzymatic activities can be modulated gradually. We give detailed protocols for the synthesis of these molecules. Summarizing our experience with different enzymes, we explain their use for fundamental studies of conformational plasticity, as well as the potential as molecular probes.
Keywords: Allosteric control; Bioluminescence probe; DNA spring; DNA-based tool; Enzyme kinetics; Mechanical control of enzymes; Mechanophores; Protein-DNA chimeras; Supramolecular constructions.
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