Site-directed Mutagenesis of Key Residues Unveiled a Novel Allosteric Site on Human Adenosine Kinase for Pyrrolobenzoxa(thia)zepinone-Non-Nucleoside Inhibitors

A novel allosteric site on human Adenosine Kinase (hAK) for pyrrolobenzoxa(thia)zepinone non-nucleoside inhibitors was hypothesized by us. Bioinformatics analysis led us to identify the key residues of the postulated site. We herein cloned and expressed specific, single and double point mutants of hAK. Gratifyingly, mutated enzymes show reduced susceptibility to our compounds, while maintaining comparable affinity for nucleoside inhibitors to the wild type enzyme. This study represents the first characterization and validation of a novel allosteric site in hAK.