A General, Label-Free and Homogeneous Electrochemical Strategy for Probing of Protease Activity and Screening of Inhibitor

Abstract

Proteases play a critical role in regulating various physiological processes from protein digestion to wound healing. Monitoring the activity of proteases and screening their inhibitors as potential drug molecules are of great importance for the early diagnosis and treatment of many diseases. In this work, we reported a general, label-free and homogeneous electrochemical method for monitoring protease activity based on the peptide–copper interaction. Cleavage of peptide substrate results in the generation of a copper-binding chelator peptide with a histidine residue in the first or third position (His1 or His3) at the N-terminal. The redox potential and current of copper coordinated with the product are different from the free copper or the copper complex with the substrate, thus allowing for the detection of protease activity. Angiotensin-converting enzyme (ACE) and thrombin were determined as the model analytes. The label-free and homogeneous electrochemical method can be used for screening protease inhibitors with high simplicity and sensitivity.