title: A Three-Enzyme Pathway with an Optimised Geometric Arrangement to Facilitate Substrate Transfer.
abstract: Cascade reactions drive and regulate a variety of metabolic activities. Efficient coupling of substrate transport between enzymes is important for overall pathway activity and also controls the depletion of intermediate molecules that drive the reaction forward. Here, we assembled a three-enzyme pathway on a series of DNA nanoscaffolds to investigate the dependence of their activities on spatial arrangement. Unlike previous studies, the overall activity of the three-enzyme pathway relied less on inter-enzyme distance and more on the geometric patterns that arranged them within a relatively small range of 10-30 nm. Pathway intermediate detection demonstrated that the assembled enzyme systems quickly depleted the intermediate molecules through efficient reaction coupling.
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ, 85281, USA.
authors.forename: Minghui
authors.initials: M
authors.lastname: Liu
authors.affiliation: Department of Chemistry, Center for Computational and Integrative Biology, Rutgers University-Camden, 315 Penn Street, Camden, NJ, 08102, USA.
authors.forename: Jinglin
authors.initials: J
authors.lastname: Fu
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ, 85281, USA.
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Qi
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ, 85281, USA.
authors.forename: Shaun
authors.initials: S
authors.lastname: Wootten
authors.affiliation: Biodesign Center for Innovations in Medicine, The Biodesign Institute, Arizona State University, 1001 S McAllister Avenue, Tempe, AZ, 85281, USA.|School of Molecular Sciences, Arizona State University, Physical Sciences Building, Room D-102, P. O. Box 871604, Tempe, AZ, 85287-1604, USA.
authors.forename: Neal W
authors.initials: NW
authors.lastname: Woodbury
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ, 85281, USA.|School of Molecular Sciences, Arizona State University, Physical Sciences Building, Room D-102, P. O. Box 871604, Tempe, AZ, 85287-1604, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Biodesign Center for Molecular Design and Biomimetics, The Biodesign Institute, Arizona State University, 1001 S. McAllister Avenue, Tempe, AZ, 85281, USA. hao.yan@asu.edu.|School of Molecular Sciences, Arizona State University, Physical Sciences Building, Room D-102, P. O. Box 871604, Tempe, AZ, 85287-1604, USA. hao.yan@asu.edu.
authors.forename: Hao
authors.initials: H
authors.lastname: Yan
mesh_terms: D002262:Carboxy-Lyases / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism*; D007770:L-Lactate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D008291:Malate Dehydrogenase / Q000737:chemistry / Q000378:metabolism; D049329:Nanostructures / Q000737:chemistry*; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D013816:Thermodynamics
chemical_list: D004798:Enzymes; D004247:DNA; D007770:L-Lactate Dehydrogenase; D008291:Malate Dehydrogenase; D002262:Carboxy-Lyases; C021916:oxaloacetate decarboxylase
keywords: DNA nanostructures; enzyme cascade; malic dehydrogenase; self-assembly