Interenzyme Substrate Diffusion for an Enzyme Cascade Organized on Spatially Addressable DNA Nanostructures
Top Cited Papers
- 13 March 2012
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 134 (12), 5516-5519
- https://doi.org/10.1021/ja300897h
Abstract
Spatially addressable DNA nanostructures facilitate the self-assembly of heterogeneous elements with precisely controlled patterns. Here we organized discrete glucose oxidase (GOx)/horseradish peroxidase (HRP) enzyme pairs on specific DNA origami tiles with controlled interenzyme spacing and position. The distance between enzymes was systematically varied from 10 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.Keywords
This publication has 24 references indexed in Scilit:
- Challenges and opportunities for structural DNA nanotechnologyNature Nanotechnology, 2011
- Folding DNA to create nanoscale shapes and patternsNature, 2006
- Water Dynamics in the Hydration Layer around Proteins and MicellesChemical Reviews, 2005
- Diffusion in BiofilmsJournal of Bacteriology, 2003
- DNA-Directed Assembly of Bienzymic Complexes from In Vivo Biotinylated NAD(P)H:FMN Oxidoreductase and LuciferaseChemBioChem, 2002
- Swinging Arms and Swinging Domains in Multifunctional Enzymes: Catalytic Machines for Multistep ReactionsAnnual Review of Biochemistry, 2000
- The Molecular Basis of Substrate ChannelingJournal of Biological Chemistry, 1999
- Kinetic Characterization of Linear Diffusion of the Restriction Endonuclease EcoRV on DNABiochemistry, 1998
- Site-to-site directed immobilization of enzymes with bis-NAD analogues.Proceedings of the National Academy of Sciences, 1983
- Metabolic Compartmentation: Symbiotic, Organellar, Multienzymic, and MicroenvironmentalAnnual Review of Microbiology, 1974