Engineered enzyme sensors based on allosteric regulation

Summary It is proposed to develop an approach for constructing enzyme based sensors for detecting biomoleculesThe sensor is an enzyme modified by site directed mutagenesis to create a binding site specific for a target biomoleculeA library of engineered enzyme sensors is constructed to serve as a repository of potential bindersThe binding site is positioned in the immediate vicinity of the enzymeandapos s active site such that the active site is allosterically affected by the binding of the biomolecule to the binding siteBinding is detected in a chemical reaction catalyzed by the enzyme using a chromogenic or fluorogenic substrateImplementation will be straightforward and as simple as adding the biomolecule of interestor a sample that contains itto the enzyme sensor and observing the outcome of the reactioni ecolor or fluorescenceNarrative The project will result in creation of a general approach to obtain enzyme based sensors capable of detecting the presence of biomolecules and automatically signaling their presenceThe approach can be used in research and developmentmolecular diagnostic testing and drug discoveryThe ease of testing allows the method to be applied broadly