Inter-enzyme substrate diffusion for an enzyme cascade organized on spatially addressable DNA nanostructures

By Jinglin Fu, Minghui Liu, Yan Liu, N. Woodbury, Hao Yan. Journal of the American Chemical Society, 2012

612 citations recorded by the source corpus.

Spatially addressable DNA nanostructures facilitate the self-assembly of heterogeneous elements with precisely controlled patterns. Here we organized discrete GOx/HRP enzyme pairs on specific DNA origami tiles with controlled inter-enzyme spacing and position. The distance between enzymes was systematically varied from 10 nm to 65 nm and the corresponding activities were evaluated. The study revealed two different distance dependent kinetic processes associated with the assembled enzyme pairs. Strongly enhanced activity was observed for those assemblies in which the enzymes were closely spaced, while the activity dropped dramatically for enzymes as little as 20 nm apart. Increasing the spacing further resulted in a much weaker distance dependence. Combined with diffusion modeling, the results suggest that Brownian diffusion of intermediates in solution governed the variations in activity for more distant enzyme pairs, while dimensionally-limited diffusion of intermediates across connected protein surfaces contributed to the enhancement in activity for closely spaced GOx/HRP assemblies. To further test the role of limited dimensional diffusion along protein surfaces, a noncatalytic protein bridge was inserted between GOx and HRP to connect their hydration shells. This resulted in substantially enhanced activity of the enzyme pair.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/dc7c3475d4af73ad65b0e59854d9bb22e4290c97

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Molecular Biology

  1. Travis A. Meyer
  2. P. K. Dutta
  3. Yonggang Ke
  4. Pengfei Wang
  5. Victor Pan

Citations in the corpus