Histone controlled aggregation of tetraphenylethene probe: A new method for the detection of protease activity

Abstract A novel fluorescence “turn-on” assay for protease activity based on controlled aggregation of a tetraphenylethene probe is reported. A tetraphenylethene derivative (TPE-2+) with two positive charges is used in this study. TPE-2+ has no emission in an aqueous solution. Heparin is an anionic polymer, which can induce aggregation of TPE-2+. Strong aggregation-induced fluorescence emission (AIE) is observed. Histone is a cationic protein, it binds with heparin via electrostatic attractive interactions, which hinders the binding of TPE-2+ to heparin. TPE-2+ is released, and a decreased emission signal is detected. Upon the addition of a protease, histone is hydrolyzed into small fragments, which weakens its competitive binding to heparin. Free heparin induces aggregation of TPE-2+. A turn-on emission signal is recorded. An efficient and convenient fluorescence assay for protease activity is therefore established.