Fluorescing Isofunctional Ribonucleosides: Assessing Adenosine Deaminase Activity and Inhibition

Abstract

The enzymatic conversion of isothiazolo[4,3-d]pyrimidine-based adenosine (^tz A) and 2-aminoadenosine (^tz 2-AA) analogues to the corresponding isothiazolo[4,3-d]pyrimidine-based inosine (^tz I) and guanosine (^tz G) derivatives is evaluated and compared to the conversion of native adenosine to inosine. Henri-Michaelis-Menten analyses provides the foundation for a high-throughput screening assay, and the efficacy of the assay is showcased by fluorescence-based analysis of ^tz A conversion to ^tz I in the presence of known and newly synthesized inhibitors.