Determination of D1 Protease Activity and Screening for Lead Compounds of Inhibitor by Capillary Zone Electrophoresis

Capillary zone electrophoresis (CZE) with UV detection has been developed for the determination of D1 protease activity and screening for lead compounds of its inhibitor. Separation conditions of oligopeptides were optimized as phosphate buffer (50 mM, pH 3. 0) at a running voltage of 18 kV, with a capillary length of 43 cm × 75 μm (i.d). Hydrolysis activity assay of D1 protease and screening of lead compounds of its inhibitor were performed by CZE. In the study, two lead compounds, ITP 21 and ITP 26, showed inhibitory efficiency of 26% and 13%, respectively, while other compounds had no inhibitory effects on D1 protease activity. This method is simple, convenient and suitable for screening studies of the inhibitors of D1 protease.