title: PNA-peptide assembly in a 3D DNA nanocage at room temperature.
abstract: Proteins and peptides fold into dynamic structures that access a broad functional landscape; however, designing artificial polypeptide systems is still a great challenge. Conversely, DNA engineering is now routinely used to build a wide variety of 2D and 3D nanostructures from hybridization based rules, and their functional diversity can be significantly expanded through site specific incorporation of the appropriate guest molecules. Here we demonstrate a new approach to rationally design 3D nucleic acid-amino acid complexes using peptide nucleic acid (PNA) to assemble peptides inside a 3D DNA nanocage. The PNA-peptides were found to bind to the preassembled DNA nanocage in 5-10 min at room temperature, and assembly could be performed in a stepwise fashion. Biophysical characterization of the DNA-PNA-peptide complex was performed using gel electrophoresis as well as steady state and time-resolved fluorescence spectroscopy. Based on these results we have developed a model for the arrangement of the PNA-peptides inside the DNA nanocage. This work demonstrates a flexible new approach to leverage rationally designed nucleic acid (DNA-PNA) nanoscaffolds to guide polypeptide engineering.
authors.affiliation: Center for Bio-Inspired Solar Fuel Production, Arizona State University, Tempe, Arizona 85287, United States.
authors.forename: Justin D
authors.initials: JD
authors.lastname: Flory
authors.forename: Sandip
authors.initials: S
authors.lastname: Shinde
authors.forename: Su
authors.initials: S
authors.lastname: Lin
authors.forename: Yan
authors.initials: Y
authors.lastname: Liu
authors.forename: Hao
authors.initials: H
authors.lastname: Yan
authors.forename: Giovanna
authors.initials: G
authors.lastname: Ghirlanda
authors.forename: Petra
authors.initials: P
authors.lastname: Fromme
mesh_terms: D004247:DNA / Q000737:chemistry*; D015394:Molecular Structure; D049329:Nanostructures / Q000737:chemistry*; D020135:Peptide Nucleic Acids / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry*; D013696:Temperature
chemical_list: D020135:Peptide Nucleic Acids; D010455:Peptides; D004247:DNA