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Selected Works of Dustin Patterson
Article
Two-dimensional crystallization of P22 virus-like particles
Journal of Physical Chemistry B (2016)
  • Hideyuki Yoshimura, Meiji University
  • Hideyuki Yoshimura, Indiana University Bloomington
  • Ethan Edwards, Indiana University Bloomington
  • Masaki Uchida, Indiana University Bloomington
  • Kimberly McCoy, Indiana University Bloomington
  • Raj Roychoudhury, Indiana University Bloomington
  • Benjamin Schwarz, Indiana University Bloomington
  • Dustin Patterson
  • Trevor Douglas, Indiana University Bloomington
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Abstract
Virus-like particles (VLPs) are well established platforms for constructing functional biomimetic materials. The VLP from the bacteriophage P22 can be used as a nanocontainer to sequester active enzymes, at high concentration, within its cavity through a process of directed self-assembly. Construction of ordered 2D assemblies of these catalytic VLPs can be envisioned as a functional membrane. To achieve this, it is important to establish methods to fabricate densely packed monolayers of VLPs. Highly ordered assemblies of P22 can also be utilized as a two-dimensional (2D) crystal for electron crystallography to get precise structural information on the VLP. Here we report 2D crystallization of different P22 morphologies: P22 procapsid (PC), enzyme encapsulated PC (β-glycosidase and enhanced green fluorescent protein), empty shell (PC without scaffold proteins, ES), the expanded form of P22 (EX), and enzyme encapsulated EX (NADH oxidase). The 2D crystals of P22 VLPs were formed on a positively charged lipid monolayer at the water–air interface with a subphase containing 1% trehalose. A P22 solution, injected underneath the lipid monolayer, floated to the surface because of the density difference between the subphase and protein solution. The lipid monolayer, with adsorbed P22, was transferred to a holey carbon grid and was examined by electron microscopy. 2D crystals were obtained from a subphase containing 100 mM NaCl, 10 mM MES (pH 5.0), and 1% trehalose. The diffraction spots from the transferred film extended to the sixth order in negatively stained samples and the 10th order in cryo-electron microscopy samples.
Keywords
  • MonolayersLipidsCrystallizationPeptides and proteinsMolecules
Disciplines
Publication Date
July 7, 2016
DOI
10.1021/acs.jpcb.6b01425
Citation Information
Hideyuki Yoshimura, Hideyuki Yoshimura, Ethan Edwards, Masaki Uchida, et al.. "Two-dimensional crystallization of P22 virus-like particles" Journal of Physical Chemistry B Vol. 120 Iss. 26 (2016) p. 5938 - 5944
Available at: http://works.bepress.com/dustin-patterson/22/