title: Assembling Enzymatic Cascade Pathways inside Virus-Based Nanocages Using Dual-Tasking Nucleic Acid Tags.
abstract: The packaging of proteins into discrete compartments is an essential feature for cellular efficiency. Inspired by Nature, we harness virus-like assemblies as artificial nanocompartments for enzyme-catalyzed cascade reactions. Using the negative charges of nucleic acid tags, we develop a versatile strategy to promote an efficient noncovalent co-encapsulation of enzymes within a single protein cage of cowpea chlorotic mottle virus (CCMV) at neutral pH. The encapsulation results in stable 21-22 nm sized CCMV-like particles, which is characteristic of an icosahedral T = 1 symmetry. Cryo-EM reconstruction was used to demonstrate the structure of T = 1 assemblies templated by biological soft materials as well as the extra-swelling capacity of these T = 1 capsids. Furthermore, the specific sequence of the DNA tag is capable of operating as a secondary biocatalyst as well as bridging two enzymes for co-encapsulation in a single capsid while maintaining their enzymatic activity. Using CCMV-like particles to mimic nanocompartments can provide valuable insight on the role of biological compartments in enhancing metabolic efficiency.
authors.affiliation: Department of Biomolecular Nanotechnology, MESA+ Institute for Nanotechnology, University of Twente , 7500 AE Enschede, The Netherlands.
authors.forename: Melanie
authors.initials: M
authors.lastname: Brasch
authors.affiliation: Department of Biomolecular Nanotechnology, MESA+ Institute for Nanotechnology, University of Twente , 7500 AE Enschede, The Netherlands.
authors.forename: Rindia M
authors.initials: RM
authors.lastname: Putri
authors.affiliation: Department of Biomolecular Nanotechnology, MESA+ Institute for Nanotechnology, University of Twente , 7500 AE Enschede, The Netherlands.
authors.forename: Mark V
authors.initials: MV
authors.lastname: de Ruiter
authors.affiliation: Department of Structure of Macromolecules, Centro Nacional de Biotecnología/CSIC , Cantoblanco, 28049 Madrid, Spain.|Centro Nacional de Microbiología/Instituto de Salud Carlos III , Majadahonda, 28220 Madrid, Spain.
authors.forename: Daniel
authors.initials: D
authors.lastname: Luque
authors.affiliation: Department of Biomolecular Nanotechnology, MESA+ Institute for Nanotechnology, University of Twente , 7500 AE Enschede, The Netherlands.
authors.forename: Melissa S T
authors.initials: MS
authors.lastname: Koay
authors.affiliation: Department of Structure of Macromolecules, Centro Nacional de Biotecnología/CSIC , Cantoblanco, 28049 Madrid, Spain.
authors.forename: José R
authors.initials: JR
authors.lastname: Castón
authors.affiliation: Department of Biomolecular Nanotechnology, MESA+ Institute for Nanotechnology, University of Twente , 7500 AE Enschede, The Netherlands.
authors.forename: Jeroen J L M
authors.initials: JJ
authors.lastname: Cornelissen
mesh_terms: D055162:Biocatalysis; D017795:Bromovirus / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism*; D009696:Nucleic Acids / Q000737:chemistry / Q000378:metabolism*; D010316:Particle Size; D010734:Phosphogluconate Dehydrogenase / Q000737:chemistry / Q000378:metabolism*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000378:metabolism*; D013499:Surface Properties
chemical_list: D009696:Nucleic Acids; D010734:Phosphogluconate Dehydrogenase; D005949:Glucose Oxidase; D017853:Phosphotransferases (Alcohol Group Acceptor); C020578:gluconokinase