Modulation of Enzymatic Reaction Trajectories via Applied Mechanical Forces

Enzymes have evolved with remarkable activities, and selectivities. This is accomplished through the intricate formation and stabilization of catalytic transition states. In many cases enzyme structures are dynamic, with subtle conformational changes potentially occurring during reactant binding, catalysis and product release. There is a growing appreciation that mechanical forces are involved in catalytic pathway trajectories and we hypothesize that that addition of applied external mechanical forces could be exploited to influence kinetic steps thus directing alternative catalytic reaction trajectories. This hypothesis will be explored using both simple DNA springs and DNA tweezers as well as more complex DNA origami tweezers and DNA origami force clamps. These tools will be used to apply tension and compression forces in and around the active site of the thermostable alcohol dehydrogenase D enzyme from Pyrococcus furiosus. These efforts will further elucidate the importance of applied mechanical forces on the biocatalytic mechanisms and will help inform efforts focused on the design and development of synthetic and biomimetic enzymes and biocatalysts.