title: Stepwise molecular display utilizing icosahedral and helical complexes of phage coat and decoration proteins in the development of robust nanoscale display vehicles.
abstract: A stepwise addition protocol was developed to display cargo using bacteriophage P22 capsids and the phage decoration (Dec) protein. Three-dimensional image reconstructions of frozen-hydrated samples of P22 particles with nanogold-labeled Dec bound to them revealed the locations of the N- and C-termini of Dec. Each terminus is readily accessible for molecular display through affinity tags such as nickel-nitrilotriacetic acid, providing a total of 240 cargo-binding sites. Dec was shown by circular dichroism to be a β-sheet rich protein, and fluorescence anisotropy binding experiments demonstrated that Dec binds to P22 heads with high (~110 nm) affinity. Dec also binds to P22 nanotubes, which are helically symmetric assemblies that form when the P22 coat protein contains the F170A amino acid substitution. Several classes of tubes with Dec bound to them were visualized by cryo-electron microscopy and their three-dimensional structures were determined by helical reconstruction methods. In all instances, Dec trimers bound to P22 capsids and nanotubes at positions where three neighboring capsomers (oligomers of six coat protein subunits) lie in close proximity to one another. Stable interactions between Dec and P22 allow for the development of robust, nanoscale size, display vehicles.
authors.affiliation: Department of Chemistry & Biochemistry, University of California-San Diego, 9500 Gilman Drive, La Jolla, CA 92093, USA.
authors.forename: Kristin N
authors.initials: KN
authors.lastname: Parent
authors.forename: Christina T
authors.initials: CT
authors.lastname: Deedas
authors.forename: Edward H
authors.initials: EH
authors.lastname: Egelman
authors.forename: Sherwood R
authors.initials: SR
authors.lastname: Casjens
authors.forename: Timothy S
authors.initials: TS
authors.lastname: Baker
authors.forename: Carolyn M
authors.initials: CM
authors.lastname: Teschke
mesh_terms: D017100:Bacteriophage P22 / Q000737:chemistry*; D008422:Materials Testing; D053769:Nanocapsules / Q000737:chemistry* / Q000648:ultrastructure; D043942:Nanotubes / Q000737:chemistry* / Q000648:ultrastructure; D019151:Peptide Library; D014677:Pharmaceutical Vehicles / Q000138:chemical synthesis*; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: D053769:Nanocapsules; D019151:Peptide Library; D014677:Pharmaceutical Vehicles; D014764:Viral Proteins