A small-molecule FRET reporter for the real-time visualization of cell-surface proteolytic enzyme functions

• Main Authors: Mu, Jing, Liu, Fang, Rajab, Muhammad Shafiq, Shi, Meng, Li, Shuang, Goh, Chiching, Lu, Lei, Xu, Qing-Hua, Liu, Bin, Ng, Lai Guan, Xing, Bengang
• Other Authors: School of Biological Sciences
• Format: Article
• Language: English
• Published: 2015
• Subjects: DRNTU::Science::Chemistry
• Online Access: https://hdl.handle.net/10356/103964; http://hdl.handle.net/10220/24640
• Institution: Nanyang Technological University

Real-time imaging of cell-surface-associated proteolytic enzymes is critical to better understand their performances in both physiological and pathological processes. However, most current approaches are limited by their complexity and poor membrane-anchoring properties. Herein, we have designed and synthesized a unique small-molecule fluorescent probe, which combines the principles of passive exogenous membrane insertion and Förster resonance energy transfer (FRET) to image cell-surface-localized furin-like convertase activities. The membrane-associated furin-like enzymatic cleavage of the peptide probe leads to an increased fluorescence intensity which was mainly localized on the plasma membrane of the furin-expressed cells. This small-molecule fluorescent probe may serve as a unique and reliable reporter for real-time visualization of endogenous cell-surfaceassociated proteolytic furin-like enzyme functions in live cells and tissues using one-photon and two-photon microscopy.