Peroxidase-like activity of [Mn2(bpmp)]3+ complex enhanced by ADP: Application in a real-time colorimetric assay for small molecule kinases

Abstract

In this research, we demonstrated ADP could enhance the peroxidase like activity the artificial peroxidase [Mn2(bpmp)]3+ complex, enhancing the oxidation of hydrogen peroxide (H2O2) and ABTS [Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt]. By integrating kinase mediated conversion of ATP to ADP and ADP-enhanced peroxidase like activity of [Mn2(bpmp)]3+ complex, a real-time colorimetric assay for small molecule kinases was constructed. This real-time colorimetric assay method was applied on quantitative assays of four different small molecule kinases. Also, this method was applied to determine the half maximal inhibitory concentration (IC50) of hexokinase inhibitors, and the relative inhibition efficiency of these inhibitors could be confirmed by the naked eye, indicating the applicability of this method to high throughput screening (HTS).