원문정보
초록
영어
Activity-based analysis of proteolytic enzymes has been considered as a key step for understanding their biological functions against numerous diseases.[1] Here, we demonstrate quantumdot (QD)-based nanosesnors to detect the protease activity. The main principle entails the bioluminescent resonance energy transfer (BRET) between Renilla luciferase as an energy donor and QD as an energy acceptor, where the metal-affinity triggers the association of the carboxyl groups of QDs with the hexahistidine tag of luciferase in close proximity.[2] By incorporating a specific peptide substrate into the C-terminus of luciferase protein in its expression, the BRET ratio of donor intensity at 480 nm to acceptor intensity at 655 nm was analyzed as a function of protease concentration in the presence of Ni2+. Upon adding the protease to the QD-luciferase nanosensor, the Ni2+-dependent BRET ratio significantly decreased with a strong emission from QDs, as a result of cleaving the peptide substrate. We anticipate that this strategy would be well suited to analyze the activities of many other proteases with high sensitivity and selectivity.