A real‐time fluorescence assay for protease activity and inhibitor screening based on the aggregation‐caused quenching of a perylene probe. (1st April 2018)

• Record Type: Journal Article
• Authors: Wang, Yan; Zhang, Zhifang; Zhang, Ya; Yu, Cong
• Is Part Of: Luminescence. Volume 33:Number 4(2018)
• Page Start: 790
• Page End: 796
• Publication Date: 2018-04-01
• Subjects: aggregation -- fluorescence -- peptide -- perylene probe -- protease activity; Luminescence -- Periodicals; Bioluminescence -- Periodicals; Chemiluminescence -- Periodicals; Luminescence -- Periodicals; 535.35
• DOI: 10.1002/bio.3478
• Languages: English
• ISSNs: 1522-7235

Abstract

Abstract: We have established a real‐time and label‐free fluorescence turn‐on strategy for protease activity detection and inhibitor screening via peptide‐induced aggregation‐caused quenching of a perylene probe. Because of electrostatic interactions and high hydrophilicity, poly‐l ‐glutamic acid sodium salt (PGA; a negatively charged peptide) could induce aggregation of a positively charged perylene probe (probe 1) and the monomer fluorescence of probe 1 was effectively quenched. After a protease was added, PGA was enzymatically hydrolyzed into small fragments and probe 1 disaggregated. The fluorescence recovery of probe 1 was found to be proportional to the concentration of protease in the range from 0 to 1 mU/ml. The detection limit was down to 0.1 mU/ml. In the presence of a protease inhibitor, protease activity was inhibited and fluorescence recovery reduced. Moreover, we demonstrated the potential application of our method in a complex mixture sample including 1% human serum. Our method is simple, fast and cost effective.