Characterization of Flavonol Inhibition of DnaB Helicase: Real-Time Monitoring, Structural Modeling, and Proposed Mechanism
Figure 6
Real-time monitoring of KpDnaB helicase activity. (a) Schematic representation of fluorescence helicase assay based on FRET. The dsDNA substrate was prepared by annealing 2 oligonucleotides, a 5′ fluorophore-labeled (Alexa Fluor 488) 22-nucleotide donor, and a 3′ quencher-labeled (BHQ1) 36-nucleotide quencher. When the dsDNA substrate is unwound by the helicase, the fluorophore (F) emits upon its release from the quencher (Q). (b) Inhibitory effects of flavonols on KpDnaB helicase activity. Myr and Gal were selected for this assay.
