Design of a Hierarchical Assembly at a Solid-Liquid Interface Using an Asymmetric Protein Needle

Design and control of processes for a hierarchical assembly of proteins remain challenging because it requires consideration of design principles with atomic-level accuracy. Previous studies have adopted symmetry-based strategies to minimize the complexity of protein???protein interactions and this has placed constraints on the structures of the resulting protein assemblies. In the present work, we used an anisotropic-shaped protein needle, gene product 5 (gp5) from bacteriophage T4 with a C-terminal hexahistidine-tag (His-tag) (gp5_CHis), to construct a hierarchical assembly with two distinct protein???protein interaction sites. High-speed atomic force microscopy (HS-AFM) measurements reveal that it forms unique tetrameric clusters through its N-terminal head on a mica surface. The clusters further self-assemble into a monolayer through the C-terminal His-tag. The HS-AFM images and displacement analyses show that the monolayer is a network-like structure rather than a crystalline lattice. Our results expand the toolbox for constructing hierarchical protein assemblies based on structural anisotropy.

Automated summary

In this study, a hierarchical protein assembly system with two distinct protein-protein interaction sites was constructed using an anisotropic-shaped protein needle. High-speed atomic force microscopy measurements showed the formation of unique tetrameric clusters through the N-terminal head of the protein on a mica surface. These clusters further self-assembled into a monolayer structure through the C-terminal His-tag. The results expand the toolbox for constructing hierarchical protein assemblies based on structural anisotropy.