title: Real-time fluorometric turn-on assay for protease activity and inhibitor screening with a benzoperylene probe.
abstract: A real-time fluorescence turn-on strategy for protease activity and inhibitor screening has been developed. A negatively charged benzo[ghi]perylene derivative (probe 1) was employed. Protamine is a cationic protein which can induce aggregation of probe 1 via strong electrostatic and hydrophobic interactions. The fluorescence of probe 1 was efficiently quenched. In the presence of a protease, protamine was enzymatically hydrolyzed and probe 1 de-aggregated. The recovery of the probe 1 monomer fluorescence could be detected. The protease activity could be monitored in real-time. In addition, upon addition of a protease inhibitor, the protease-catalyzed hydrolysis was inhibited, which led to a decreased fluorescence recovery. The fluorometric assay thus could also be employed for screening protease inhibitors.
authors.affiliation: Department of Chemistry, Zhejiang Sci-Tech University, Hangzhou 310018, P. R. China. zhujintao1968@zstu.edu.cn.
authors.forename: Chuibei
authors.initials: C
authors.lastname: Zhou
authors.forename: Wenying
authors.initials: W
authors.lastname: Li
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Meiding
authors.initials: M
authors.lastname: Yang
authors.forename: Yang
authors.initials: Y
authors.lastname: Li
authors.forename: Jintao
authors.initials: J
authors.lastname: Zhu
authors.forename: Cong
authors.initials: C
authors.lastname: Yu
mesh_terms: D003199:Computer Systems; D004353:Drug Evaluation, Preclinical / Q000379:methods; D004789:Enzyme Activation / Q000502:physiology; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000737:chemistry*; D005470:Fluorometry / Q000379:methods*; D010569:Perylene / Q000031:analogs & derivatives* / Q000737:chemistry; D014357:Trypsin / Q000032:analysis* / Q000378:metabolism
chemical_list: D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; C006706:benzoperylene; D010569:Perylene; D014357:Trypsin