Strategies and Applications for Supramolecular Protein Self-assembly.

Supramolecular chemistry achieves higher-order molecular self-assembly through non-covalent interactions. Utilizing supramolecular methods to explore the polymorphism of proteins, the building blocks of life, from a "bottom-up" perspective is essential for constructing diverse and functional biomaterials. In recent years, significant progress has been achieved in the design strategies and functional applications of supramolecular protein self-assembly, becoming a focal point for researchers. This paper reviews classical supramolecular strategies driving protein self-assembly, including electrostatic interactions, metal coordination, hydrogen bonding, hydrophobic interactions, host-guest interactions, and other mechanisms. We discuss how these supramolecular interactions regulate protein assembly processes and highlight protein supramolecular assemblies' unique structural and functional advantages in constructing artificial photosynthetic systems, protein hydrogels, bio-delivery systems, and other functional materials. The enormous potential and significance of supramolecular protein materials are elucidated. Finally, the challenges in preparing and applying protein supramolecular assemblies are summarized, and future development directions are projected.

Current Advances of Atomically Dispersed Metal-Centered Nanozymes for Tumor Diagnosis and Therapy.

Nanozymes, which combine enzyme-like catalytic activity and the biological properties of nanomaterials, have been widely used in biomedical fields. Single-atom nanozymes (SANs) with atomically dispersed metal centers exhibit excellent biological catalytic activity due to the maximization of atomic utilization efficiency, unique metal coordination structures, and metal-support interaction, and their structure-activity relationship can also be clearly investigated. Therefore, they have become an emerging alternative to natural enzymes. This review summarizes the examples of nanocatalytic therapy based on SANs in tumor diagnosis and treatment in recent years, providing an overview of material classification, activity modulation, and therapeutic means. Next, we will delve into the therapeutic mechanism of SNAs in the tumor microenvironment and the advantages of synergistic multiple therapeutic modalities (e.g., chemodynamic therapy, sonodynamic therapy, photothermal therapy, chemotherapy, photodynamic therapy, sonothermal therapy, and gas therapy). Finally, this review proposes the main challenges and prospects for the future development of SANs in cancer diagnosis and therapy.

Morphological Transformation between Orthogonal Dynamic Covalent Self-Assembly of Imine-Boroxine Hybrid Polymer Nanocapsules and Thin Films via Linker Exchange.

Orthogonal dynamic covalent self-assembly is used as a facile method for constructing polymer hollow nanocapsules (NCs) and thin films. The bifunctional precursor 4-formylphenylboronic acid is symmetrically installed with a boronic acid group for the boroxine linkage, and an aldehyde group for the Schiff base reaction which can react with twofold symmetry linkers ethylenediamine and para phenylenediamine to attain polymer NCs and nanosheets. Owing to the reversibility of the imine linkages, the mutual morphological transformation between polymer NCs and thin films via an amine-imine-exchange strategy is successfully achieved. Multiple reversible covalent bonds allow the control the release of the load in polymer NCs using different techniques. This may be useful for designing stimulus-responsive smart materials.

Recent development in the design of artificial enzymes through molecular imprinting technology.

Enzymes, a class of proteins or RNA with high catalytic efficiency and specificity, have inspired generations of scientists to develop enzyme mimics with similar capabilities. Many enzyme mimics have been developed in the past few decades based on small molecules, DNA, and nanomaterials. These artificial enzymes are of great interest because of their low cost and high stability. However, most of these enzyme mimics do not have the desired substrate selectivity. The substrate selectivity of natural enzymes usually stems from a specific binding pocket. A powerful method to create substrate binding cavities is molecular imprinting technology (MIT). Molecularly imprinted polymers (MIPs) have three main characteristics: structural predictability, identification specificity, and application versatility compared with other identification systems. The MIP-based artificial enzymes have the advantages of simple preparation, low cost, and high stability and can realize excellent catalytic activity and selectivity. The development of MIP-based artificial enzymes has been further promoted by optimization methods such as imprinting transition state molecules, post-imprinting modification, opening cross-linked polymers' internal space, and some special preparation methods. Combining molecular imprinting technology with nanozymes, the synergistic effect of both solved the defect of lack of specificity of nanozymes and improved their catalytic activity. This paper summarizes the recent research progress in preparing high-performance artificial enzymes based on MIPs and molecularly imprinted nanozymes. We hope to provide a reference for the design of artificial enzymes, reduce the gap between artificial enzymes and natural enzymes, and thus broaden the application of artificial enzymes in human life and production.

"On/Off" Switchable Sequential Light-Harvesting Systems Based on Controllable Protein Nanosheets for Regulation of Photocatalysis.

A controllable protein nanostructures-based "On/Off" switchable artificial light-harvesting system (LHS) with sequential multistep energy transfer and photocatalysis was reported herein for mimicking the natural LHS in both structure and function. Single-layered protein nanosheets were first constructed via a reversible covalent self-assembly strategy using cricoid stable protein one (SP1) as building blocks to realize an ordered arrangement of pigments. Fluorescent chromophores like carbon dots (CDs) can be precisely distributed on the protein nanosheets superficially via electrostatic interactions and make the ratio between donors and acceptors adjustable. After being anchored with a photocatalysis center (eosin-5-isothiocyanate, EY), the constructed LHS could sequentially transfer energy between two kinds of chromophores (CD1 and CD2), and further transfer to EY center with a high efficiency of 84%. Interestingly, the Förster resonance energy transfer (FRET) process of our LHS could be reversibly "On/Off" switched by the redox regulated assembly and disassembly of SP1 building blocks. Moreover, the LHS has been further proved to promote the yield of a model cross-coupling hydrogen evolution reaction and regulate the process of the reaction with the FRET process "On/Off" state.

Hierarchical protein self-assembly into dynamically controlled 2D nanoarrays via host-guest chemistry.

A dynamically reversible two-dimensional (2D) protein assembly system was designed based on host-guest interactions and was triggered to disassemble via a competition mechanism. The artificially tunable and reversible protein assembly architectures hold great potential for on/off switches in bio-systems.

Biocompatible Diselenide-Containing Protein Hydrogels with Effective Visible-Light-Initiated Self-Healing Properties.

Smart hydrogels are typical functional soft materials, but their functional and mechanical properties are compromised upon micro- or macro-mechanical damage. In contrast, hydrogels with self-healing properties overcome this limitation. Herein, a dual dynamic bind, cross-linked, self-healing protein hydrogel is prepared, based on Schiff base bonds and diselenide bonds. The Schiff base bond is a typical dynamic covalent bond and the diselenide bond is an emerging dynamic covalent bond with a visible light response, which gives the resulting hydrogel a dual response in visible light and a desirable self-healing ability. The diselenide-containing protein hydrogels were biocompatible due to the fact that their main component was protein. In addition, the hydrogels loaded with glucose oxidase (GOx) could be transformed into sols in glucose solution due to the sensitive response of the diselenide bonds to the generated hydrogen peroxide (H2O2) by enzymatic catalysis. This work demonstrated a diselenide-containing protein hydrogel that could efficiently self-heal up to nearly 100% without compromising their mechanical properties under visible light at room temperature.

Construction of Ultralarge Two-Dimensional Fluorescent Protein Arrays via a Reengineered Rhodamine B-Based Molecular Tool.

The self-luminous property of enhanced green fluorescent protein (EGFP) makes it an extremely attractive building block for creating functional biomaterials. A practical challenge in the design of EGFP-based materials, however, stems from the structural and chemical heterogeneity of the EGFP surface. In this study, a maleimide-functionalized rhodamine B molecule (RhG2M) was designed as a versatile molecular tool to overcome this obstacle. Site-specific modification of an EGFP variant (EGFP-4C) with RhG2M allowed for the fabrication of a series of well-defined two-dimensional (2D) arrays that span nano- and micrometer scales. Furthermore, the resulting ultralarge 2D EGFP-4C arrays feature both structural uniformity and flexibility, together with the inherent optical properties, making them advanced materials with great potential for practical applications. In addition, this strategy can be further extended into three dimensions and applied to the modular generation of periodic functional materials with more complex structures.

Difunctionalized pillar[5]arene-based polymer nanosheets for photodynamic therapy of Staphylococcus aureus infection.

Bacterial infections pose severe threats to global public health security. Developing antibacterial agents with both high efficiency and safety to handle this problem has become a top priority. Here, highly stable and effective polymer nanosheets have been constructed by the covalent co-assembly of a pillar[5]arene derivative and metalloporphyrin for photodynamic antibacterial therapy (PDAT). The monolayer nanosheets are strongly positively charged and thus capable of binding with Staphylococcus aureus (SA) through electrostatic interactions. Additionally, the nanosheets can be activated to generate reactive oxygen species (ROS) under white-light irradiation, and exhibit satisfactory antibacterial performance towards SA. More importantly, cell viability assays demonstrate that the nanosheets show little to no cytotoxicity impact on mammalian cells even when the concentrations are much higher than those employed in the antibacterial studies. The above results suggest that the polymer nanosheets could be an effective antibacterial agent to overcome bacterial infections and hold a broad range of potential applications in real life.

Rational Design and Biological Application of Antioxidant Nanozymes.

Nanozyme is a type of nanostructured material with intrinsic enzyme mimicking activity, which has been increasingly studied in the biological field. Compared with natural enzymes, nanozymes have many advantages, such as higher stability, higher design flexibility, and more economical production costs. Nanozymes can be used to mimic natural antioxidant enzymes to treat diseases caused by oxidative stress through reasonable design and modification. Oxidative stress is caused by imbalances in the production and elimination of reactive oxygen species (ROS) and reactive nitrogen species (RNS). This continuous oxidative stress can cause damage to some biomolecules and significant destruction to cell structure and function, leading to many physiological diseases. In this paper, the methods to improve the antioxidant properties of nanozymes were reviewed, and the applications of nanozyme antioxidant in the fields of anti-aging, cell protection, anti-inflammation, wound repair, cancer, traumatic brain injury, and nervous system diseases were introduced. Finally, the future challenges and prospects of nanozyme as an ideal antioxidant were discussed.

Light-responsive vesicles for enantioselective release of chiral drugs prepared from a supra-amphiphilic M-helix.

A kind of light-responsive vesicle was prepared by aqueous self-assembly of α-CD and an azobenzene-containing M-helical foldamer, which displayed dynamic disassembly-reassembly structural transformation when alternately irradiated by UV and visible light. Distinctively, this vesicle also exhibited enantioselective release abilities toward racemic propranolol (a ß-blocker), owing to the M-helical building blocks.

Cucurbit[8]uril-based supramolecular nanocapsules with a multienzyme-cascade antioxidative effect.

A supramolecular nanocapsule was constructed by the ternary host-guest complexation of azobenzene (Azo) and methylviologen (MV) to cucurbit[8]uril (CB[8]) and the subsequent self-assembly. The supramolecular nanocapsule with both glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities can mimic the intracellular enzymatic reactive oxygen species (ROS) defense system.

A remote optically controlled hydrolase model based on supramolecular assembly and disassembly of its enzyme-like active site.

A photoresponsive hydrolase model was constructed through the spatial organization of histidine/arginine-containing peptide supra-amphiphiles that are held together by cucurbit[8]uril (CB[8]) methylviologen (MV) azobenzene (Azo) ternary complexation and subsequently self-assemble into highly uniform giant vesicles. The reversible morphological transition of the vesicular structures to non-assembled peptide fragments was triggered by azobenzene photoisomerization. This enables the assembly/disassembly of its enzyme-like active site to cause a dramatic change in hydrolytic activity. The dynamic process can be directly monitored to determine the supramolecular structure-related enzymatic parameters, which may help to understand how the regulation of enzyme activity is coupled to the aggregation behaviors of natural enzymes.

An ultrathin iron-porphyrin based nanocapsule with high peroxidase-like activity for highly sensitive glucose detection.

For the first time, an ultrathin iron-porphyrin based polymer nanocapsule with multiple peroxidase-like catalytic centers was constructed by covalently assembling iron-porphyrin monomers; this nanocapsule with a single molecule thickness shell acted as a highly efficient artificial enzyme for mimicking peroxidase. On the basis of the peroxidase-like activity of Fe-TPyP based nanocapsules (Fe-TPyP NCs), a highly sensitive colorimetric sensor for glucose determination was fabricated, the limit of detection was found to be as low as 0.098 µM. This study provided a novel strategy for developing artificial enzymes based on covalently assembled nanostructures. Furthermore, the colorimetric sensor for glucose determination showed potential applications in biomedicine and biology.

Reductive-Responsive, Single-Molecular-Layer Polymer Nanocapsules Prepared by Lateral-Functionalized Pillar[5]arenes for Targeting Anticancer Drug Delivery.

Herein, a new reductive-responsive pillar[5]arene-based, single-molecule-layer polymer nanocapsule is constructed for drug delivery. The functionalized system shows good biocompatibility, efficient internalization into targeted cells and obvious triggered release of entrapped drugs in a reducing environment such as cytoplasm. Besides, this smart vehicle loaded with anticancer drug shows excellent inhibition for tumor cell proliferation and exhibits low side effect on normal cells. This work not only demonstrates the development of a new reductive-responsive single molecular layer polymer nanocapsule for anticancer drug targeting delivery but also extends the design of smart materials for biomedical applications.