EcoRV catalysis with a pre-bent substrate

Enzymes are deformable molecules which often adapt their conformation to the substrate’s geometry. In the case of restriction enzymes acting on DNA, the substrate (DNA) is deformable also. While it is well established that external mechanical stress exerted on an enzyme modulates the enzymatic activity, the effect of mechanically stressing the substrate is less explored. Here we present the restriction enzyme EcoRV with a series of pre-bent DNA substrates and observe modulation of the overall speed of the enzymatic reaction by up to a factor 50. While in all cases the reaction is slowed down compared to the unstressed substrate, we observe peaks in the reaction rate as we vary the state of stress of the DNA substrate. Both bending moments and torsional moments on the DNA substrate affect the reaction rate.