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Protein-incorporated soft networks have received remarkable attention during the past several years. They possess desirable properties similar to native tissues and organs and exhibit unique advantages in applications. However, fabrication of protein-based hydrogels usually suffers from complex protein mutation and modification or chemical synthesis, which limited the scale and yield of production. Meanwhile, the lack of rationally designed noncovalent interactions in networks may result in a deficiency of the dynamic features of materials. Therefore, a highly efficient method is needed to include supramolecular interactions into protein hydrogel to generate a highly dynamic hydrogel possessing integrated tissue-like properties. Here, we report the design and construction of native protein-based supramolecular synthetic protein hydrogels through a simple and efficient one-pot polymerization of acrylamide and ligand monomers in the presence of a ligand-binding protein. The supramolecular interactions in the network yield integrated dynamic properties, including remarkable stretchability over 10,000% of their original length, ultrafast self-healing abilities within 3-4 s, tissue-like fast stress relaxation, satisfactory ability of adhesion to different living and nonliving substrates, injectability, and high biocompatibility. Furthermore, this material demonstrated potential as a biosensor to monitor small finger movements. This strategy provides a new avenue for fabricating synthetic protein hydrogels with integrated features.

The cellular compartmentation induced by self-assembly of natural proteins has recently attracted widespread attention due to its structural-functional significance. Among them, as a highly conserved metabolic enzyme and one of the potential targets for cancers and parasitic diseases in drug development, CTP synthase (CTPS) has also been reported to self-assemble into filamentous structures termed cytoophidia. To elucidate the dynamical mechanism of cytoophidium filamentation, we utilize single-molecule fluorescence imaging to observe the real-time self-assembly dynamics of CTPS and the coordinated assembly between CTPS and its interaction partner, Δ 1 -pyrroline-5-carboxylate synthase (P5CS). Significant differences exist in the direction of growth and extension when the two proteins self-assemble. The oligomer state distribution analysis of the CTPS minimum structural subunit under different conditions and the stoichiometry statistics of binding CTPS and P5CS by single-molecule fluorescence photobleach counting further confirm that the CTPS cytoophidia are mainly stacked with tetramers. CTPS can act as the nucleation core to induce the subsequent growth of the P5CS filaments. Our work not only provide evidence from the molecular level for the self-assembly and coordinated assembly (coassembly) of CTPS with its interaction partner P5CS in vitro but also offer new experimental perspectives for the dynamics research of coordinated regulation between other protein polymers.

In biology, polymorphism is a well-known phenomenon by which a discrete biomacromolecule can adopt multiple specific conformations in response to its environment. This term can be extended to the ability of biomacromolecules to pack into different ordered patterns. Thus, exploration and control of the polymorphism of biomacromolecules via supramolecular methods have been key steps in achieving bioinspired structures, developing bioinspired functional materials, and exploring the mechanisms of these self-assembly processes, which are models for more complex biological systems. This task could be difficult for proteins and carbohydrates due to the complicated multiple noncovalent interactions of these two species which can hardly be manipulated.In this account, dealing with the structural polymorphisms from biomacromolecular assemblies, we will first briefly comment on the problems that carbohydrate/protein assemblies are facing, and then on the basis of our long-term research on carbohydrate self-assemblies, we will summarize the new strategies that we have developed in our laboratory in recent years to explore and control the polymorphism of carbohydrate/protein assemblies.Considering the inherent ability of carbohydrates to recognize lectin, we proposed the "inducing ligand" strategy to assemble natural proteins into various nanostructures with highly ordered packing patterns. The newly developed inducing ligand approach opened a new window for protein assembly where dual noncovalent interactions (i.e., carbohydrate-protein interactions and dimerization of rhodamine) instead of the traditionally used protein-protein interactions direct the assembly pattern of proteins. As a result, various polymorphisms of protein assemblies have been constructed by simply changing the ligand chemical structure and/or the rhodamine dimerization.Another concept that we proposed for glycopolymer self-assembly is DISA (i.e., deprotection-induced glycopolymer self-assembly). It is well known that protection-deprotection chemistry has been employed to construct complex oligosaccharide structures. However, its application in glycopolymer self-assembly has been overlooked. We initiated this new strategy with diblock copolymers. Such copolymers with a carbohydrate block having protected pendent groups exist as single chains in organic media. The self-assembly can be initiated by the deprotection of the pendent groups. The process was nicely controlled by introducing various protective groups with different deprotection rates. Later on, the DISA process has been proven practical in water and even in the cellular environment, which opens a new avenue for the development of polymeric glycomaterials.Finally, the resultant polymeric glyco-materials, as a new type of biomimetic materials, provide a nice platform for investigating the functions of glycocalyx. The glycocalyx-mimicking nanoparticles achieved unprecedent functions which exceed their carbohydrate precursors. Here, the reversion of tumor-associated macrophages induced by glycocalyx-mimicking nanoparticles will be discussed with potential applications in cancer immunotherapy, where such a reversion effect could be combined with other methods (e.g., tumor checkpoint blockade).

Polymorphism has been the subject of investigation across different research disciplines. In biology, polymorphism could be interpreted in such a way that discrete biomacromolecules can adopt diversiform specific conformations/packing arrangement, and this polymorph-dependent property is essential for many biochemical processes. For example, bacterial flagellar filament, composed of flagellin, switches between different supercoiled state allowing the bacteria to swim and tumble. However, in artificial supramolecular systems, it is often challenging to achieve polymorph control and prediction, and in most cases, two or more concomitant polymorphs of similar formation energies coexist. Here, we show that a tetrameric protein with properly oriented binding sites on its surface can arrange into diverse protein tubes with distinct helical parameters by adding specifically designed inducing ligands. We examined several parameters of the ligand that would influence the protein tube formation and found that the flexibility of the ligand linker and the dimerization pose of the ligand complex is critical for the successful production of the tubes and eventually influence the specific helical polymorphs of the formed tubes. A surface lattice accommodation model was further developed to rationalize the geometrical relationship between each helical tube type. Molecular simulation was used to elucidate the interactions between ligands and SBA and molecular basis for polymorphic switching of the protein tubes. Moreover, the kinetics of structural formation was studied and the ligand design was found that can affect the kinetics of the protein polymerization pathway. In short, our designed protein tubes serves as an enlightening system for understanding how a protein polymer composed of a single protein switches among different helical states.