2026-10-15T04:24:39Z https://www.scidb.cn/oai

10.57760/sciencedb.07439 2023-02-15T15:52:58Z

2023-02-15 Dipolar coupling-based electron paramagnetic resonance method for protease enzymatic characterization and inhibitor screening doi:10.57760/sciencedb.07439 en Herein, we report an EPR-based method for protease enzymatic characterization and inhibitor screening. This method utilizes dual paramagnetically-labeled probes consisting of a nitroxide spin probe and a Gd3+ ion flanking a peptide that could be specifically cleaved by protease caspase-3. Distance-dependent dipolar coupling between the two paramagnetic centers can be modulated by the protease cleavage activity, thus providing a straightforward and convenient method for protease activity detection using EPR spectroscopy under ambient conditions. Moreover, time-course monitoring of the protease catalyzed cleavage reaction demonstrated that this EPR-based method could not only allow a direct quantitative enzymatic kinetic assessment, but also could be used for protease inhibitor screening, thus holding great potential in drug discovery studies. Caspase-3; Electron Paramagnetic Rssonance; Inhibitor Scerrning; Protease enzymatic YuLu Liu Aokun Kuang jian Lu yi Tian Changlin Guo Xiaoqi Zhang binbo PUBLIC https://creativecommons.org/licenses/by/4.0/ dataset http://www.doi.org/10.1039/d1cc03301h Science Data Bank