Recognized as the primary RNA virus to be categorized and extensively studied, the tobacco mosaic virus (TMV) is foundational to advancements in virology, molecular biology, and biomaterials. Over the past few decades, the deep comprehension of the TMV coat protein (TMVcp) molecular structure and assembly principles has stimulated a surge in research on TMVcp structural design using genetic engineering and chemical modification techniques. The unique characteristics of TMVcp, including its nanoscale orderly structure, ease of modification, and considerable drug loading capacity, have enabled significant progress in its biomedical applications. This review summarizes the advanced strategies deployed for TMVcp design and multidimensional assembly and underscores the prototypical applications of TMVcp‐based biomaterials in bioimaging, drug delivery, tissue engineering, and biosensing. Ultimately, the future prospects of TMVcp research in structural design and biomedical applications are explored, which encompass artificial intelligence‐guided structural and functional design, the development of stimulus‐responsive biomaterials, and potential clinical translation.

Engineering noncovalent interactions for assembling nonspherical proteins into supramolecular architectures with tunable morphologies and dynamics is challenging due to the structural heterogeneity and complexity of protein surfaces. Herein, we employed an anisotropic building block l-rhamnulose-1-phosphate aldolase (RhuA) to control supramolecular polymorphism in highly ordered protein assemblies by introducing histidine residues. Histidine-based π-π stacking interactions enabled thermodynamically controlled self-organization of RhuA to form three-dimensional (3D) nanoribbons and crystals. Self-assembly of different 3D crystal phases was kinetically modulated by the strong metal ion-histidine chelation, and double-helical protein superstructures were formed by engineering increased histidine interactions at the RhuA binding surface. Their structural properties and dynamics were determined via fluorescence microscopy, transmission electron microscopy, atomic force microscopy, and small-angle X-ray scattering. This work is aimed at expanding the toolbox for the programming of tunable, highly ordered, protein superstructures and increasing the understanding of the mechanisms of protein interfacial interactions.

Self-assembly guided by biological molecules is a promising approach for fabricating predesigned nanostructures. Protein is one such biomolecule possessing deterministic 3D crystal structure and peptide information, which acts as a good candidate for templating functional nanoparticles (fNPs). However, inadequate coordination efficacy during the establishment of interfacial interactions with fNPs makes it highly challenging to precisely fabricate designed nanostructures and functional materials. Here, a facile and robust strategy is reported for the hierarchical assembly of fNPs into ordered architectures, with unprecedentedly large sizes up to tens of micrometers, using a hollow cylinder-shaped tobacco mosaic virus coat protein (TMV disk). The rational design of the site-specific functional groups on the TMV disk not only demonstrates the powerful capability of directing various discrete fNP assemblies with high controllability but also assists in precise assembly of a TMV monolayer sheet structure for further organizing homogeneous and heterogeneous fNP periodic lattices by varying the types of fNPs. The high precision and adjustability of the pattern fashions of different fNPs unambiguously corroborate the validity of this innovative strategy, which provides a convenient route to design and assemble protein-based hierarchical ordered architectures for use in nanophotonics and nanodevices.

The science of protein self-assembly has experienced significant development, from discrete building blocks of self-assembled nanoarchitectures to advanced nanostructures with adaptive functionalities. Despite the prominent achievements in the field, the desire of designing de novo protein-nanoparticle (NP) complexes and constructing dynamic NP systems remains highly challenging. In previous works, l-rhamnulose-1-phosphate aldolase (C98RhuA) tetramers were self-assembled into two-dimensional (2D) lattices via disulfide bond interactions. These interactions provided 2D lattices with high structural quality and a sophisticated assembly mode. In this study, we devised a rational design for RhuA building blocks to fabricate 2D functionalized protein lattices. More importantly, the lattices were used to direct the precise assembly of NPs into highly ordered and diverse nanoarchitectures. These structures can be employed as an excellent tool to adequately verify the self-assembly mode and structural quality of the designed RhuA crystals. The subsequent redesign of RhuA building blocks enabled us to predictably produce a novel protein lattice whose conformational dynamics can be controllably regulated. Thus, a dynamic system of AuNP lattices was achieved. Transmission electron microscopy and small-angle X-ray scattering indicated the presence of these diverse NP lattices. This contribution enables the fabrication of future NP structures in a more programmable manner with more expected properties for potential applications in nanoelectronics and other fields.