Protein‐Based Patterning to Spatially Functionalize Biomimetic Membranes

Abstract

The bottom‐up reconstitution of proteins for their modular engineering into synthetic cellular systems can reveal hidden protein functions in vitro. This is particularly evident for the bacterial Min proteins, a paradigm for self‐organizing reaction‐diffusion systems that displays an unexpected functionality of potential interest for bioengineering: the directional active transport of any diffusible cargo molecule on membranes. Here, the MinDE protein system is reported as a versatile surface patterning tool for the rational design of synthetically assembled 3D systems. Employing two‐photon lithography, microswimmer‐like structures coated with tailored lipid bilayers are fabricated and demonstrate that Min proteins can uniformly pattern bioactive molecules on their surface. Moreover, it is shown that the MinDE system can form stationary patterns inside lipid vesicles, which allow the targeting and distinctive clustering of higher‐order protein structures on their inner leaflet. Given their facile use and robust function, Min proteins thus constitute a valuable molecular toolkit for spatially patterned functionalization of artificial biosystems like cell mimics and microcarriers.

Protein‐Assisted Large‐Scale Assembly and Differential Patterning of DNA Origami Lattices

“…The exact concentration of MinD and MinE required for a specific pattern can vary depend-ing on the substrate, membrane composition, and buffer conditions, but a multitude of patterns can be achieved in a variety of sample conditions. [38][39][40][41] Thus, we set out to characterize the different patterns and the resulting origami lattices. For this, we used a custom ImageJ macro to extract the mean percentage of area available for DNA origami superstructure formation (Figure 3c), as well as the mean width of lattice segments or diameter in the case of the inverse spot pattern (Figure 3d).…”