DEEPLY QUENCHED ENZYME SENSORS AND BINDING SENSORS

摘要

Sensors for detecting enzyme activity are provided that include a substrate module comprising a substrate for the enzyme of interest and a fluorescent label, a quencher, and a detection module. The detection module binds to the substrate module either before or after the enzyme acts on the substrate and sequesters the label from the quencher, resulting in an increased signal from the label. Sensors for detecting protein-protein interactions are also provided that include a quencher and a labeled first polypeptide. Binding of the first polypeptide to a second polypeptide sequesters the label from the quencher, resulting in an increased signal from the label. Methods using the sensors to detect enzyme activity and to screen for compounds affecting enzyme activity or to detect protein-protein interactions and to screen for compounds affecting protein-protein interactions, respectively, are also described.