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.

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.

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.

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.

Prediction of the potentially suitable areas of Leonurus japonicus in China based on future climate change using the optimized MaxEnt model.

Leonurus japonicus Houtt. is a traditional Chinese medicinal plant with high medicinal and edible value. Wild L. japonicus resources have reduced dramatically in recent years. This study predicted the response of distribution range of L. japonicus to climate change in China, which provided scientific basis for the conservation and utilization. In this study, 489 occurrence points of L. japonicus were selected based on GIS technology and spThin package. The default parameters of MaxEnt model were adjusted by using ENMeva1 package of R environment, and the optimized MaxEnt model was used to analyze the distribution of L. japonicus. When the feature combination in the model parameters is hing and the regularization multiplier is 1.5, the MaxEnt model has a higher degree of optimization. With the AUC of 0.830, our model showed a good predictive performance. The results showed that L. japonicus were widely distributed in the current period. The maximum temperature of warmest month, the min temperature of coldest month, the precipitation of wettest month, the precipitation of driest month, and altitude were the main environmental factors affecting the distribution of L. japonicus. Under the three climate change scenarios, the suitable distribution area of L. japonicus will range shift to high latitudes, indicating that the distribution of L. japonicus has a strong response to climate change. The regional change rate is the lowest under the SSP126-2090s scenario and the highest under the SSP585-2090s scenario.

Shape-Anisotropic Assembly of Protein Nanocages with Identical Building Blocks by Designed Intermolecular π-π Interactions.

Protein lattices that shift the structure and shape anisotropy in response to environmental cues are closely coupled to potential functionality. However, to design and construct shape-anisotropic protein arrays from the same building blocks in response to different external stimuli remains challenging. Here, by a combination of the multiple, symmetric interaction sites on the outer surface of protein nanocages and the tunable features of phenylalanine-phenylalanine interactions, a protein engineering approach is reported to construct a variety of superstructures with shape anisotropy, including 3D cubic, 2D hexagonal layered, and 1D rod-like crystalline protein nanocage arrays by using one single protein building block. Notably, the assembly of these crystalline protein arrays is reversible, which can be tuned by external stimuli (pH and ionic strength). The anisotropic morphologies of the fabricated macroscopic crystals can be correlated with the Å-to-nm scale protein arrangement details by crystallographic elucidation. These results enhance the understanding of the freedom offered by an object's symmetry and inter-object π-π stacking interactions for protein building blocks to assemble into direction- and shape-anisotropic biomaterials.

Facilitating Pro-survival Mitophagy for Alleviating Parkinson's Disease via Sequence-Targeted Lycopene Nanodots.

The pathogenesis of Parkinson's disease is closely linked to impaired mitochondrial function and abnormal mitophagy. Biocompatible natural antioxidants effectively protect dopaminergic neurons. However, the main challenge in using natural antioxidants for Parkinson's disease therapy is creating a delivery platform to achieve neuron-targeted enrichment. Herein, we synthesized rationally sequence-targeted lycopene nanodots using recombinant human H-ferritin nanocages with lycopene loading into the cavity and lipophilic triphenylphosphonium (TPP) coupling on the outer surface. The nanodots allow for the neural enrichment and mitochondrial regulation of lycopene through blood-brain barrier transcytosis and neuronal mitochondria-targeting capability. These anti-ROS nanodots protect neuronal mitochondrial function and promote PINK1/Parkin-mediated mitophagy in MPTP toxicity-induced neurons in vivo and in vitro, which favors the secretory efflux of pathogenic α-synuclein and the survival of dopaminergic neurons. Moreover, these nanodots restore the Parkinson-like motor symptoms in Parkinson's model mice. This noninvasive sequence-targeted delivery strategy with excellent biocompatibility for pro-survival mitophagy-mediated pathology alleviation makes it a promising approach for treating and preventing Parkinson's disease.

N-acetyl-L-leucine protects MPTP-treated Parkinson's disease mouse models by suppressing Desulfobacterota via the gut-brain axis.

Parkinson's disease (PD) is the second most common neurodegenerative disease, and communication between the gut and brain (the gut-brain axis) has been found to be essential in behavior and cognitive function. However, the exact mechanisms underlying microbiota dysbiosis in PD progression have not yet been elucidated. Our study aimed to investigate the correlation between gut microbiota disturbances and feces metabolic disorders in PD. We used 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to induce PD models and observed mice's motor symptoms, dopaminergic (DA) neuron death, and gastrointestinal dysfunction. To identify alterations in microbiota and metabolome, feces were collected from mice and analyzed using 16 S ribosomal RNA sequencing feces metabolomics. Pearson analysis was utilized to investigate correlations between the abundances of gut microbiota components and the levels of gut microbiota metabolites, displaying their interaction networks. Our findings revealed a significant increase in Desulfobacterota in the PD mouse model and 151 differentially expressed fecal metabolites between PD and vehicle mice. Moreover, Pearson correlation analysis suggested that the protective factor N-acetyl-L-leucine (NALL) may be associated with neuroinflammation in the striatum and substantia nigra, which also had a negative relationship with the concentration of Desulfobacterota. Additionally, we found that oral administration of NALL alleviated MPTP-induced Motor Impairments and DA neuronal deficits. All in all, we concluded that the decrease of NALL might lead to a significant increase of Desulfobacterota in the MPTP model mouse and subsequently result in the damage of DA neurons via the gut-brain aix pathway.

Highly Stretchable, Transparent, and Adhesive Double-Network Hydrogel Dressings Tailored with Fish Gelatin and Glycyrrhizic Acid for Wound Healing.

It remains challenging to fabricate highly stretchable and adhesive hydrogel dressings for wound healing using simple, safe, and green methods. Herein, inspired by the main components of snail mucus, a fully physical double-network (DN) hydrogel dressing composed of fish gelatin (FGel) and glycyrrhizic acid (GL) was fabricated, in which FGel provided a protein scaffold to mimic snail mucus proteins, while GL mimicked the adhesion and bioactivity of snail mucus because of its abundant carboxyl and hydroxyl groups and intrinsic immunomodulatory activity. As expected, the obtained FGel/GL hydrogel dressings exhibited outstanding mechanical and adhesive performances (flexibility, stretchability, adhesive ability, and removability), high transparency, and good antifreezing properties. More importantly, they also possessed excellent biocompatibility, cell migration, and angiogenesis ability in vitro experiments. Finally, animal experiments in vivo indicated that the FGel/GL hydrogel dressings significantly promoted full-thickness wound healing, including promoting granulation tissue formation, collagen deposition, and skin angiogenesis and inhibiting the inflammatory response. All these findings indicated that the FGel/GL hydrogel dressings have great potential for applications in the clinical treatment of wound healing.

Prediction of potential distribution areas and priority protected areas of Agastache rugosa based on Maxent model and Marxan model.

Agastache rugosa (Fisch. & C. A. Mey.) Kuntze has been widely studied because of its high medicinal and edible value. Establishing the priority protected area of wild A. rugosa can provide scientific basis for the protection of germplasm resources. In this study, we predicted the potential suitability distribution area of A. rugosa under the current and future climate scenarios with the MaxEnt model, and the dominant climate factors affecting the distribution of A. rugosa were analyzed. Based on the above results, we predicted the priority protected areas of A. rugosa with the Marxan model. The results showed that A. rugosa is mainly distributed in the eastern and central regions of China at present. In future, the suitable area of A. rugosa will increase, otherwise a few areas will shrink back and migrate to the high latitude areas as a whole. Hydrothermal conditions are the main environmental factors affecting the distribution of A. rugosa. The priority protected areas of A. rugosa are mainly distributed in Chongqing, eastern Sichuan, southern Guizhou, western Hunan and Hubei and southwestern Shaanxi, which are basically consistent with the highly suitable areas predicted by Maxent model. The results of this study are of great significance for the protection and rational utilization of species of Agastache.

Reassembly Design of Ferritin Cages.

The ferritin family is distributed in nearly all organisms and protects them from iron-induced oxidative damage. Besides, its highly symmetrical structure and biochemical features make it an appealing material for biotechnological applications, such as building blocks for multidimensional assembly, templates for nano-reactors, and scaffolds for encapsulation and delivery of nutrients and drugs. Moreover, it is of great significance to construct ferritin variants with different properties, size, and shape to further broaden its application. In this chapter, we present a routine process of the ferritin redesign and the characterization method of the protein structure to provide a feasible scheme.