Assembling Enzymatic Cascade Pathways inside Virus-Based Nanocages Using Dual-Tasking Nucleic Acid Tags

• Published in: Journal of the American Chemical Society Vol. 139; no. 4; pp. 1512 - 1519
• Main Authors: Brasch, Melanie, Putri, Rindia M, de Ruiter, Mark V, Luque, Daniel, Koay, Melissa. S. T, Castón, José R, Cornelissen, Jeroen J. L. M
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 01-02-2017
• Subjects: Biocatalysis; Bromovirus - chemistry; Bromovirus - enzymology; Bromovirus - metabolism; Glucose Oxidase - chemistry; Glucose Oxidase - metabolism; Nucleic Acids - chemistry; Nucleic Acids - metabolism; Particle Size; Phosphogluconate Dehydrogenase - chemistry; Phosphogluconate Dehydrogenase - metabolism; Phosphotransferases (Alcohol Group Acceptor) - chemistry; Phosphotransferases (Alcohol Group Acceptor) - metabolism; Surface Properties
• ISSN: 0002-7863; 1520-5126
• DOI: 10.1021/jacs.6b10948

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.