Metals at the Helm: Revolutionizing Protein Assembly and Applications.

Protein assembly is an essential process in biological systems, where proteins self-assemble into complex structures with diverse functions. Inspired by the exquisite control over protein assembly in nature, scientists have been exploring ways to design and assemble protein structures with precise control over their topologies and functions. One promising approach for achieving this goal is through metal coordination, which utilizes metal-binding motifs to mediate protein-protein interactions and assemble protein complexes with controlled stoichiometry and geometry. Metal coordination provides a modular and tunable approach for protein assembly and de novo structure design, where the metal ion acts as a molecular glue that holds the protein subunits together in a specific orientation. Metal-coordinated protein assemblies have shown great potential for developing functional metalloproteinase, novel biomaterials and integrated drug delivery systems. In this review, an overview of the recent advances in protein assemblies benefited from metal coordination is provided, focusing on various protein arrangements in different dimensions including protein oligomers, protein nanocage and higher-order protein architectures. Moreover, the key metal-binding motifs and strategies used to assemble protein structures with precise control over their properties are highlighted. The potential applications of metal-mediated protein assemblies in biotechnology and biomedicine are also discussed.

Both Hemocyanin and ß-1,3-Glucan-Binding Protein from the Shrimp Shell of Litopenaeus vannamei Are Responsible for Its Color Change from Brown to Red during Thermal Processing.

Because of the composition and structural complexity of crustacean shells, their color change mechanism during thermal processing remains unclear. This study identified and characterized two intrinsic protein components, hemocyanin (Lv-Hc) and ß-1,3-glucan-binding protein (Lv-BGBP) from Litopenaeus vannamei shrimp shells by a combination of ion-exchange chromatography, gel filtration, and mass spectrometry. It was found that a mixture of Lv-Hc, a gray protein, and Lv-BGBP (which is a natural astaxanthin-binding protein with a red color) is responsible for the brown color of fresh shrimp shells. Upon heating to 100 °C, the mixture of these proteins turned red, mimicking the color change observed in cooked shrimp shells. This transition is attributed to the extremely high thermal stability of Lv-BGBP, which has the ability to protect astaxanthin from thermal induced degradation. These findings provide significant insights into the molecular mechanism governing shrimp shell coloration, advancing our understanding of crustacean biochemistry.

Prediction of potential invasion of two weeds of the genus Avena in Asia under climate change based on Maxent.

Avena sterilis L. (A. sterilis) and Avena ludoviciana Dur. (A. ludoviciana) are extremely invasive weeds with strong competitive ability and multiple transmission routes. Both species can invade a variety of dryland crops, including wheat, corn, and beans. Asia, as the world's major food-producing continent, will experience significant losses to agricultural production if it is invaded by these weeds on a large scale. This study used the MaxEnt model and ArcGIS to map the distribution of suitable habitats of the two species in Asia under climate change conditions. The constructed model comprised four levels, with a total of 25 index-level indicator factors used to evaluate the invasion risk of the two species. The results showed that the distribution of suitable habitats for both Avena species was highly dependent on precipitation and temperature. Under climate warming conditions, although overall the total suitable area is predicted to decrease compared to the current period, there are still moderately or highly suitable areas. Asian countries need to provide early warning for areas with significant increases in moderate and highly suitable zones for these two species of weeds under the background of climate change. If there is already an invaded area or if the suitability of the original area is increased, this should be closely monitored, and control measures should be taken to prevent further spread and deterioration.

The potential habitat of Angelica dahurica in China under climate change scenario predicted by Maxent model.

Since the 20th century, global climate has been recognized as the most important environmental factor affecting the distribution of plants. Angelica dahurica (A. dahurica) has been in great demand as a medicinal herb and flavoring, but the lack of seed sources has hindered its development. In this study, we utilized the Maxent model combined with Geographic Information System (GIS) to predict the potential habitat of A. dahurica in China based on its geographical distribution and 22 environmental factors. This prediction will serve as a valuable reference for the utilization and conservation of A. dahurica resources.The results indicated that: (1) the Maxent model exhibited high accuracy in predicting the potential habitat area of A. dahurica, with a mean value of the area under the ROC curve (AUC) at 0.879 and a TSS value above 0.6; (2) The five environmental variables with significant effects were bio6 (Min temperature of the coldest month), bio12 (Annual Precipitation), bio17 (Precipitation of Driest Quarter), elevation, and slope, contributing to a cumulative total of 89.6%. Suitable habitats for A. dahurica were identified in provinces such as Yunnan, Guizhou, Guangxi, Sichuan, Hunan, and others. The total area of suitable habitat was projected to increase, with expansion primarily in middle and high latitudes, while areas of decrease were concentrated in lower latitudes. Under future climate change scenarios, the centers of mass of suitable areas for A. dahurica were predicted to shift towards higher latitudes in the 2050s and 2090s, particularly towards the North China Plain and Northeast Plain. Overall, it holds great significance to utilize the Maxent model to predict the development and utilization of A. dahurica germplasm resources in the context of climate change.

Construction of An Artificial Photosynthesis System with A Single CdS QDs-Ferritin Hybrid Molecule.

Establishing artificial photosynthesis systems in a simple but effective manner to mitigate the greenhouse effect and address the energy crisis remains challenging. The combination of photocatalysis technology with bioengineering is an emerging field with great potential to construct such artificial photosynthesis systems, but so far, it has barely been explored in this area. Herein, an artificial photocatalysis platform is constructed with high CO2 conversion and H2O splitting capability by integration of CdS QDs into the intra-subunit interface of H-type ferritin (Marsupenaeus japonicus), a natural ferroxidase through protein interface redesign. The crystal structure of the synthesized CdS QDs with engineered ferritin molecules as bio-templates confirmed the design at an atomic level. Notably, upon absorbing desirable visible light (≈420 nm), such a single CdS-ferritin hybrid molecule is able to selectively catalyze the reduction of CO2 into HCOOH (≈90%), meanwhile catalyzing the oxidation of H2O into O2 in an aqueous environment. The O2 production rate reached to 180 µmol g-1 h-1, and the HCOOH output hit almost 800 µmol g-1 h-1. This work advances the utilization of the four-helix bundle structure for crafting artificial photosynthesis systems, facilitating the seamless integration of bioengineering and photocatalysis technology.

Construction of robust protein nanocage by designed disulfide bonds for active cargo molecules protection in the gastric environment.

Notwithstanding the progress made, cargo molecules encapsulated within ferritin via oral administration in the gastric environment remains a persistent challenge. This study focuses on the strategic enhancement of ferritin stability in harsh gastric environment. By taking advantagie of computational-assisted design, we strategically introduced up to 96 disulfide bonds along three key inter-subunit interfaces to one single ferritin molecule with human H-chain ferritin and shrimp (Marsupenaeus japonicus) ferritin as starting materials, producing two kinds of robust ferritin nanocages with markedly enhanced acid and protease (pepsin and rennin) resistance. The crystal structure of ferritin nanocage confirmed our design at an atomic level. Encapsulation experiments demonstrated successful loading of bioactive cargo molecules (e.g., doxorubicin) into the engineered ferritin nanocages, with pronouncedly improved protection against leakage under acidic condition and the presence of pepsin and rennin as compared to their native counterparts. This study presents a potential approach for the design and engineering of protein nanocages for oral administration.

Comparison of recombinant protein Z with natural protein Z derived from malt: From structure to functional properties.

Protein Z (PZ) is a prominent albumin found in the endosperm of barley seeds with a molecular weight of approximately 40 kDa. Its multifaceted functional attributes, including trypsin- and thrombin-inhibiting bioactivities and superior foaming properties, have garnered significant attention in research. Considering the post-translational modifications of PZ natural in barley malt, we tried to express recombinant protein Z (rPZ) in E. coli. The present study aims to undertake a comparative analysis between natural PZ and rPZ in order to elucidate their respective characteristics. After spectral analysis, there are significant differences in their secondary and tertiary structures. In addition, rPZ showed superior foamability and foam stability. As for the serpin-like activity, the inhibition rate of rPZ is much higher than that of PZ. In contrast with the inhibition activity, the digestability of rPZ is much lower than that of PZ. As for the cargo carrier properties, rPZ showed an excellent ability to stabilize astaxanthin at 37 °C. These results suggest that rPZ is more suitable as protein carrier, due to the high foamability, serpin-like activity and low digestive stability, which not only give a brief view of recombinant protein, but also give a direction for PZ in cargo delivery.

Vertasile ferritin nanocages: Applications in detection and bioimaging.

Food safety and human health remain significant concerns in the food industry. Detecting food contaminants and diagnosing diseases are critical aspects. Ferritin, an iron storage protein widely found in nature, offers unique advantages. Its hollow protein nanocage structure, distinct interfaces, hydrophobic or hydrophilic channels, and B-C loop regions recognized by transferrin receptor 1 make ferritin versatile for detecting heavy metals, free radicals, and bioimaging both in vitro and in vivo. This review summarizes ferritin's general characteristics, its specific properties as biosensors, and its applications in food safety and in vivo imaging. It emphasizes not only ferritin's role in detecting heavy metals like mercury and chemical hazards but also its potential in early diagnosing chronic diseases such as tumors, macrophages, and kidney diseases. Further research into ferritin promises advancements in enhancing food safety and improving human health diagnostics.

Understanding the Role of Filamentous Actin in Food Quality: From Structure to Application.

Actin, a multifunctional protein highly expressed in eukaryotes, is widely distributed throughout cells and serves as a crucial component of the cytoskeleton. Its presence is integral to maintaining cell morphology and participating in various biological processes. As an irreplaceable component of myofibrillar proteins, actin, including G-actin and F-actin, is highly related to food quality. Up to now, purification of actin at a moderate level remains to be overcome. In this paper, we have reviewed the structures and functions of actin, the methods to obtain actin, and the relationships between actin and food texture, color, and flavor. Moreover, actin finds applications in diverse fields such as food safety, bioengineering, and nanomaterials. Developing an actin preparation method at the industrial level will help promote its further applications in food science, nutrition, and safety.

A comprehensive review on hemocyanin from marine products: Structure, functions, its implications for the food industry and beyond.

Hemocyanin, an oxygen-transport protein, is widely distributed in the hemolymph of marine arthropods and mollusks, playing an important role in their physiological processes. Recently, hemocyanin has been recognized as a multifunctional glycoprotein involved in the immunological responses of aquatic invertebrates. Consequently, the link between hemocyanin functions and their potential applications has garnered increased attention. This review offers an integrated overview of hemocyanin's structure, physicochemical characteristics, and bioactivities to further promote the utilization of hemocyanin derived from marine products. Specifically, we review its implication in two aspects of food and aquaculture industries: quality and health. Hemocyanin's inducible phenoloxidase activity is thought to be an inducer of melanosis in crustaceans. New anti-melanosis agents targeted to hemocyanin need to be explored. The red-color change observed in shrimp shells is related to hemocyanin, affecting consumer preferences. Hemocyanin's adaptive modification in response to the aquatic environment is available as a biomarker. Additionally, hemocyanin is endowed with bioactivities encompassing anti-microbial, antiviral, and therapeutic activities. Hemocyanin is also a novel allergen and its allergenic features remain incompletely characterized.

Potential habitat areas and priority protected areas of Tilia amurensis Rupr in China under the context of climate change.

Introduction: Tilia amurensis Rupr (T. amurensis) is one endangered and national class II key protected wild plant in China. It has ornamental, material, economic, edible and medicinal values. At present, the resources of T. amurensis are decreasing, and the prediction of the distribution of its potential habitat in China can provide a theoretical basis for the cultivation and rational management of this species. Methods: In this study, the R language was used to evaluate 358 distribution records and 38 environment variables. The MaxEnt model was used to predict the potential distribution areas of T. amurensis under the current and future climate scenarios. The dominant environmental factors affecting the distribution of T. amurensis were analyzed and the Marxan model was used to plan the priority protected areas of this species. Results: The results showed that Bio18, Slope, Elev, Bio1, Bio9 and Bio2 were the dominant environmental factors affecting the distribution of T. amurensis. Under the future climatic scenarios, the potential suitable areas for T. amurensis will mainly distribute in the Northeast China, the total suitable area will reduce compared with the current climate scenarios, and the general trend of the centroid of suitable habitat will be towards higher latitudes. The SPF value of the best plan obtained from the priority conservation area planning was 1.1, the BLM value was 127,616, and the priority conservation area was about 57.61×104 km2. The results suggested that climate, soil and topographic factors jointly affected the potential geographical distribution of T. amurensis, and climate and topographic factors had greater influence than soil factors. Discussion: The total suitable area of T. amurensis in China under different climate scenarios in the future will decrease, so more effective protection should be actively adopted.

Growth Process of Fe-O Nanoclusters with Different Sizes Biosynthesized by Protein Nanocages.

All protein-directed syntheses of metal nanoclusters (NCs) and nanoparticles (NPs) have attracted considerable attention because protein scaffolds provide a unique metal coordination environment and can adjust the shape and morphology of NCs and NPs. However, the detailed formation mechanisms of NCs or NPs directed by protein templates remain unclear. In this study, by taking advantage of the ferritin nanocage as a biotemplate to monitor the growth of Fe-O NCs as a function of time, we synthesized a series of iron NCs with different sizes and shapes and subsequently solved their corresponding three-dimensional atomic-scale structures by X-ray protein crystallography and cryo-electron microscopy. The time-dependent structure analyses revealed the growth process of these Fe-O NCs with the 4-fold channel of ferritin as nucleation sites. To our knowledge, the newly biosynthesized Fe35O23Glu12 represents the largest Fe-O NCs with a definite atomic structure. This study contributes to our understanding of the formation mechanism of iron NCs and provides an effective method for metal NC synthesis.

Prediction of potential suitable habitats in the 21st century and GAP analysis of priority conservation areas of Chionanthus retusus based on the MaxEnt and Marxan models.

Chionanthus retusus (C. retusus) has a high economic and medicinal value, but in recent years it has been included in the list of China's major protected plants and China's Red List of Biodiversity due to the serious destruction of its wild germplasm resources. Based on 131 sample points of C. retusus, this study simulated potential habitats and spatial changes of C. retusus in the 21st century using the Maxent model combined with the geographic information system ArcGIS, predicted prioritized protected areas by the Marxan model, and assessed current conservation status through GAP analysis. The results showed that (1) when the regularization multiplier was 1.5 and the feature combinations were linear, quadratic, and fragmented, the area under the curve of the subjects in the training and test sets were both above 0.9, the true skill statistic value was 0.80, and the maximum Kappa value was 0.62, meaning that the model had high accuracy; (2) Temperature seasonality, annual precipitation, min temperature for coldest month, and precipitation of wettest month had relatively strong influences on species' ranges. (3) The moderately and optimally suitable habitats of C. retusus were primly located in the areas of southwestern Shanxi, central Hebei, western Henan, Shandong, Shaanxi, Anhui and Hubei; (4) Under different future climate scenarios, the area of each class of suitable habitat will increase for varied amounts compared to the current period, with a general trend of expansion to the south; (5) The C. retusus priority protected areas were mainly located in most of Shandong, southern Liaoning, southwestern Shanxi, western Henan, and central Hebei, and its conservation vacancy area was relatively large compared to its protected area. These results will provide scientific strategies for implementing long-term conservation of C. retusus in China and similar regions under warming conditions in the 21st century.

Can ecological niche models be used to accurately predict the distribution of invasive insects? A case study of Hyphantria cunea in China.

In recent decades, ecological niche models (ENMs) have been widely used to predict suitable habitats for species. However, for invasive organisms, the prediction accuracy is unclear. In this study, we employed the most widely used maximum entropy (MaxEnt) model and ensemble model (EM) Biomod2 and verified the practical effectiveness of the ENM in predicting the distribution areas of invasive insects based on the true occurrence of Hyphantria cunea in China. The results showed that when only limited data of invasive areas were used, the two ENMs could not effectively predict the distribution of suitable habitats of H. cunea, although the use of global data can greatly improve the prediction accuracy of ENMs. When analyzing the same data, Biomod2's prediction accuracy was significantly better than that of MaxEnt. For long-term predictions, the area of suitable habitat predicted by the ENMs was much greater than the occurrence area; for short-term predictions, the accuracy of the predicted area was significantly improved. Under the current conditions, the area of suitable habitat for H. cunea in China is 118 × 104 km2, of which 59.32% is moderately or highly suitable habitat. Future climate change could significantly increase the suitable habitat area of H. cunea in China, and the predicted area of suitable habitats in all climate scenarios exceeded 355 × 104 km2, accounting for 36.98% of the total land area in China. This study demonstrates the use of ENMs to study invasive insects and provides a reference for the management of H. cunea in China.

Structural Insights into the Reaction between Hydrogen Peroxide and Di-iron Complexes at the Ferroxidase Center of Ferritin.

The Fe(II) oxidation mechanism in the ferroxidase center of heavy chain ferritin has been studied extensively. However, the actual production of H2O2 was found to be substantially lower than expected at low flux of Fe(II) to ferritin subunits. Here, we demonstrated that H2O2 could interact with the di-iron nuclear center, leading to the production of hydroxyl radicals and oxygen. Two reaction intermediates were captured in the ferroxidase center by using the time-lapse crystallographic techniques in a shellfish ferritin. The crystal structures revealed the binding of H2O2 as a µ -1,2-peroxo-diferric species and the binding of O2 to the diferric structure. This investigation sheds light on the reaction between the di-iron nuclear center and H2O2 and provides insights for the exploitation of metalloenzymes.

Interactions between paramyosin and actin greatly improve their thermostability and gel properties.

BACKGROUND: Myofibrillar proteins, the main contributors to the quality of meat products, are the main structural protein component of muscle and have functional properties such as the formation of a 3D protein gel network and water binding. The susceptibility of meat-derived proteins to heat-induced aggregation is the functional constraint that hinders their applications in industry, and so establishing an effective but simple method to improve their thermostability of the proteins is of great importance. RESULTS: In the present study, we describe an easy approach to perform high colloidal thermostability of both paramyosin and actin by mixing them at low ionic strength. The improvement in thermal stability was found to be derived from intermolecular interactions between these two different proteins through non-covalent binding with each other. Consequently, such interactions protected each of them from thermal-induced degradation compared to individual components. Notably, this binary native protein mixture rather than single paramyosin or actin component has the ability to form protein hydrogels with a shear-thinning and reversible sol-gel transformation behavior, which is markedly different from most of reported heat-induced, denatured protein hydrogels. CONCLUSION: The present study not only presents a facile and effective strategy for improvement of the thermal stability and gel properties of a binary paramyosin and actin mixture, but also enhances our understanding of how mutual interactions of protein components affect their physicochemical and functional properties. © 2023 Society of Chemical Industry.

Improvement of physicochemical properties of lycopene by the self-assembly encapsulation of recombinant ferritin GF1 from oyster (Crassostrea gigas).

BACKGROUND: Lycopene (LYC), a carotenoid found in abundance in ripe red fruits, exhibits higher singlet oxygen quenching activity than other carotenoids. However, the stability of LYC is extremely poor due to its high double-bond content. In this paper, a nano-encapsulation strategy based on highly stable marine-derived ferritin GF1 nanocages was used to improve the thermal stability and oxidation resistance of LYC, thereby boosting its functional effectiveness and industrial applicability. RESULTS: The preparation of GF1-LYC nanoparticles benefited from the pH-responsive reversible self-assembly of GF1 to capture LYC molecules into GF1 cavities with a LYC-to-protein ratio of 51 to 1. After the encapsulation of the LYC, the reassembled GF1 nanocages maintained intact morphology and good monodispersity. The GF1-LYC nanoparticles incorporated the characteristic LYC peaks in spectrograms, and their powder form contained the crystalline form of LYC. Molecular docking revealed that LYC bound with the inner triple-axis channel areas of GF1, interacting with VAL139, LYS72, LYS65, TYR69, PHE129, HIS133, HIS62, and TYR134 amino acids through hydrophobic bonds. Fourier transform infrared spectroscopy also demonstrated the bonding of GF1 and LYC. In comparison with free LYC, GF1 reduced the thermal degradation of encapsulated LYC at 37 °C significantly and maintained the 2,2-Diphenyl-1-picrylhydrazyl (DPPH)-scavenging ability of LYC. CONCLUSION: As expected, the water solubility, thermal stability, and antioxidant capacity of encapsulated LYC from GF1-LYC nanoparticles was notably improved in comparison with free LYC, indicating that the shell-like marine ferritin nanoplatform might enhance the stable delivery of LYC and promote its utilization in the field of food nutrition and in other industries. © 2023 Society of Chemical Industry.

Chicoric acid inserted in protein Z cavity exhibits higher stability and better wound healing effect under oxidative stress.

Oxidative stress is one of the limiting factors that inhibit wound healing. Phytochemicals especially chicoric acid have the potential to act as an antioxidant and scavenge reactive oxygen species, thereby promoting wound healing. However, most of the phytochemicals were easy to be degraded during storage or using due to the oxidative status in wound site. Herein, we introduce a high stable protein Z that can encapsulate chicoric acid during foaming. TEM results showed that the size of protein Z-chicoric acid is in the range of nanoscale (named PZ-CA nanocomposite), and protein Z encapsulation can significantly improve the stability of chicoric acid under oxidative stress. Moreover, PZ-CA nanocomposite exhibited favorable antioxidant properties, biocompatibility, and the ability to promote cell migration in vitro. The role of PZ-CA nanocomposite in skin regeneration was explored by a mice model. Results in vivo suggest that the PZ-CA nanocomposite promotes wound healing with a faster rate as compared with a commercial spray solution, mostly through attenuating the oxidative stress, promoting cell proliferation and collagen deposition. This work not only provides a delivery vector for bioactive molecules, but also develops a kind of nanocomposite with the property of promoting wound healing.

Preparation and Unique Three-Dimensional Self-Assembly Property of Starfish Ferritin.

The structure and assembly properties of ferritin derived from aquatic products remain to be explored. Constructing diverse three-dimensional (3D) protein architectures with the same building blocks has important implications for nutrient delivery, medicine and materials science. Herein, ferritin from Asterias forbesii (AfFer) was prepared, and its crystal structure was resolved at 1.91 Å for the first time. Notably, different from the crystal structure of other reported ferritin, AfFer exhibited a BCT lattice arrangement in its crystals. Bioinspired by the crystal structure of AfFer, we described an effective approach for manufacturing 3D porous, crystalline nanoarchitectures by redesigning the shared protein interface involved in different 3D protein arrays. Based on this strategy, two 3D superlattices of body-centered tetragonal and simple cubicwere constructed with ferritin molecules as the building blocks. This study provided a potentially generalizable strategy for constructing different 3D protein-based crystalline biomaterials with the same building blocks.

Characterization and Structural Analyses of Enolase from Shrimp (Litopenaeus vannamei).

Transcriptome analysis had recognized enolase from shrimp Litopenaeus vannamei (L. vannamei), which is termed LvEnolase, as one of the allergens, but its amino acid sequence and protein structure have been lacking. In this study, natural LvEnolase was isolated from L. vannamei and characterized for the first time. The full-length cDNA sequence of LvEnolase was effectively cloned, which encoded 434 amino acid residues. The crystal structure of LvEnolase was successfully determined at a resolution of 2.5 Å by X-ray crystallography (PDB: 8UEL). Notably, it was observed that near the active center, a loop exists in either an open or closed state, and the open loop was associated with the product release phase. Furthermore, enzyme activity assays were conducted to validate the catalytic capabilities of purified LvEnolase. These findings significantly enhance our comprehension of the enolase family and provide valuable support for delving into the functions and characteristics of LvEnolase.