Fabrication of massive sheets of single layer patterned arrays using lipid directed reengineered phi29 motor dodecamer - PubMed (original) (raw)

Fabrication of massive sheets of single layer patterned arrays using lipid directed reengineered phi29 motor dodecamer

Feng Xiao et al. ACS Nano. 2009.

Abstract

The bottom-up assembly of patterned arrays is an exciting and important area in current nanotechnology. Arrays can be engineered to serve as components in chips for a virtually inexhaustible list of applications ranging from disease diagnosis to ultra-high-density data storage. Phi29 motor dodecamer has been reported to form elegant multilayer tetragonal arrays. However, multilayer protein arrays are of limited use for nanotechnological applications which demand nanoreplica or coating technologies. The ability to produce a single layer array of biological structures with high replication fidelity represents a significant advance in the area of nanomimetics. In this paper, we report on the assembly of single layer sheets of reengineered phi29 motor dodecamer. A thin lipid monolayer was used to direct the assembly of massive sheets of single layer patterned arrays of the reengineered motor dodecamer. Uniform, clean and highly ordered arrays were constructed as shown by both transmission electron microscopy and atomic force microscopy imaging.

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Figures

Figure 1

Figure 1

Multilayer versus single layer sheet arrays of phi29 motor dodecamer. (A) Side view of a multiple layer dodecamer array showing the horizontal face-up and face-down arrangements and the vertical head-to-tail alignment which leads to multiple layers overlap. (B) Side view of a single layer dodecamer array displaying an alternating face-up and face-down arrangement. (C) Native phi29 motor dodecamer (inset) assembled into ordered multiple layer structures as shown by negative-stain electron micrograph. (D) The negative-stain electron micrograph of reengineered phi29 motor dodecamer (inset) arrays shows that a single layer sheet was formed. (E) Projection density map of the single layer of motor dodecamers and the Fourier transform (inset). The unit cell is rectangular, with a lattice constant of ∼20 nm. The alternate orientations of the dodecamer can be observed. (F) AFM image of N-strep dodecamer arrays and a line scan across crystalline area with lattice defects (inset). The height difference between the top dodecamer layer and mica surface is ∼7.5 nm, which corresponds to a single dodecamer layer.

Figure 2

Figure 2

Negative-stain electron micrographs and corresponding fast Fourier transform (FFT) of two-layer patterned sheets of N-strep dodecamer. (A) Self-assembled huge flat sheets piled into 3D stacks. (B) Representative fast Fourier transformation. (C) The red and blue circles in the image suggested two layers sheets slightly arranged in different angle, suggesting that each sheet formed or grew independently and stacked together.

Figure 3

Figure 3

Lipid-directed formation of single layer dodecamer arrays. (A) Schematic illustration of experimental approaches. (1) Streptavidin/N-strep dodecamer solution is placed in a Teflon well; (2) biotinylated lipids DPPE and helper lipids egg PC were spread at the air−water interface to attract dodecamers to the surface of the liquid; (3) dodecamers bound to the lipid via specific biotin−streptavidin interactions. (B) Negative-stain TEM image of a single layer array produced with the aid of a lipid monolayer. (C) Fourier transforms and (D) corresponding Fourier projection maps of lipid-directed N-strep dodecamer arrays.

Figure 4

Figure 4

High magnification AFM images of self-assembled arrays of reengineered motor dodecamers. (A) Tetragonal arrays of N-strep dodecamer. (B and C) Cross-sections along the axes of the two-dimensional array. The unit cell is a parallelogram with cell dimensions of 16 nm × 13 nm. (D) Tetragonal arrays of C-strep dodecamer. The unit cell is rectangular with a lattice constant of ∼18 nm.

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