Aggregation-induced emission for the detection of peptide ligases with improving ligation efficiency

Abstract

BackgroundMonitoring peptide ligase activity is of great significance for biological research, medical diagnosis, and drug discovery. The current methods for the detection of peptide ligases suffer from the limitations of high background signal, elaborate design of substrate, and high reversibility of ligation reaction. In this work, we proposed a simple and sensitive method for ligase detection with reducing ligation reversibility on the basis of aggregation-induced emission (AIE) mechanism. ResultsThe peptide probes labeled with AIE luminogens (AIEgens) were water-soluble and emitted weak fluorescence. After ligation reaction, the enzymatic products with AIEgens showed high hydrophobicity and could readily assembly into aggregates, thus lighting up the fluorescence. More interestingly, the formation of aggregates pushed the equilibrium to the generation of the desired ligation products, thus improving the catalytic efficiency by driving the reaction towards completion. The ligation reaction conversion rate (>80 %) is significantly higher than that without blocking the reversibility with additional treatment. With sortase A (SrtA) as the analyte example, the detection limit of this method was found to be 0.01 nM with a linear range of 0–50 nM. The system was applied to evaluate the inhibition efficiency of berberine chloride and quercetin and determine the activity of SrtA in serum, lysate and Staphylococcus aureus with satisfactory results. SignificanceThis study indicated that the ligation efficiency and detection sensitivity can be improved by reducing ligation reversibility through AIE phenomenon. The proposed strategy could be used for the detection of other peptide ligases by adopting sequence-specific peptide substrates.