Methods and kits for quantifying adenosine-containing molecules within an aqueous composition. Particularly, the methods utilize a competitive fluorescence-polarization and FRET-based assays that directly measure the production of adenosine-containing compounds, particularly S-adenosylhomocysteine (AdoHcy) compounds produced by MTases. The generation of AdoHcy can be quantified by displacing a novel fluorescent probe comprising a 5-carboxytetramethylrhodamine fluorophore covalently bound to an adenosine scaffold from a catalytically inert 5-methylthioadenosine nucleosidase (MTAN) variant. One or more of the reaction materials can be pre-loaded into wells within multi-well reaction plates as a kit, which can be used to determine the enzymatic activity of MTases or conduct drug screening for potential inhibitors in a high-throughput format. Additionally, the developed assay is applicable to S-adenosyl methionine-dependent and adenosine triphosphate-dependent enzymes by detecting adenosine and various adenosine-containing molecules including 5-methylthioadenosine, adenosine monophosphate and adenosine diphosphate produced during the course of a chemical reaction.

Methods and kits for quantifying adenosine-containing molecules within an aqueous composition. Particularly, the methods utilize a competitive fluorescence-polarization and FRET-based assays that directly measure the production of adenosine-containing compounds, particularly S-adenosylhomocysteine (AdoHcy) compounds produced by MTases. The generation of AdoHcy can be quantified by displacing a novel fluorescent probe comprising a 5-carboxytetramethylrhodamine fluorophore covalently bound to an adenosine scaffold from a catalytically inert 5-methylthioadenosine nucleosidase (MTAN) variant. One or more of the reaction materials can be pre-loaded into wells within multi-well reaction plates as a kit, which can be used to determine the enzymatic activity of MTases or conduct drug screening for potential inhibitors in a high-throughput format. Additionally, the developed assay is applicable to S-adenosyl methionine-dependent and adenosine triphosphate-dependent enzymes by detecting adenosine and various adenosine-containing molecules including 5-methylthioadenosine, adenosine monophosphate and adenosine diphosphate produced during the course of a chemical reaction.