Venom Proteins Research Articles

Proteomic and functional characterisation of Trimeresurus popeiorum (Pope's pit viper) venom proteins: Role of enzymatic and non-enzymatic venom toxins in envenomation pathophysiology.

Snakebite remains a significant public health issue in tropical regions, with 4.5 to 5.4 million incidents annually. Trimeresurus popeiorum (Pope's Pit Viper), found in Southeast Asia and northeast India, poses a potential threat, yet its venom's protein composition and toxicity are poorly understood. In this study, we used label-free quantitative proteomics to analyze the venom of T. popeiorum, identifying 106 proteins across 12 venom protein families. Notably, 60 % of the venom consisted of proteolytic enzymes, correlating with its prominent metalloprotease, fibrin(ogen)lytic, procoagulant, and thrombin-like activities. The proteome composition also correlates with the clinical effects such as consumption coagulopathy and local effects, seen in victims of Pit Viper envenomation in northeast India. Our findings suggest that T. popeiorum venom is less toxic than other Viperinae species such as Daboia russelii and Echis carinatus, likely due to isoform-level variations in certain toxin classes, including metalloprotease and serine protease. The venom's lethal dose (LD50) in Swiss albino mice was 1 mg/kg, and it caused haemorrhage, tissue necrosis, edema, myotoxicity, and defibrinogenation. Histopathological examination of the TPV-treated mice showed notable toxic effects, including marked hepatic vacuolation in the liver, damage to cardiac muscle and vascular congestion in the heart, bronchial epithelial hyperplasia with cellular infiltration in the interstitial and peribronchiolar regions of the lungs, as well as tubular necrosis and haemorrhage in the kidneys. This research provides the first comprehensive analysis of T. popeiorum venom, highlighting its pharmacological effects and the need for greater medical attention to this lesser-known species.

A genus-wide study on venom proteome variation and phospholipase A2 inhibition in Asian lance-headed pit vipers (genus: Trimeresurus)

High molecular weight proteins are present abundantly in viperid venoms. The amino acid sequence can be highly variable, contributing to the structure and function diversity of snake venom protein. However, this variability remains poorly understood in many species. The study investigated the venom protein variability in a distinct clade of Asian pit vipers (Trimeresurus species complex) through comparative proteomics, applying gel electrophoresis (SDS-PAGE), liquid chromatography-tandem mass spectrometry (LCMS/MS), and bioinformatic approaches. The proteomes revealed a number of conserved protein families, within each are variably expressed protein paralogs that are unrelated to the snake phylogeny and geographic origin. The expression levels of two major enzymes, i.e., snake venom serine proteinase and metalloproteinase, correlate weakly with procoagulant and hemorrhagic activities, implying co-expression of other functionally versatile toxins in the venom. The phospholipase A2 (PLA2) abundance correlates strongly with its enzymatic activity, and a unique phenotype was discovered in two species expressing extremely little PLA2. The commercial mono-specific antivenom effectively neutralized the venoms' procoagulant and hemorrhagic effects but failed to inhibit the PLA2 activities. Instead, the PLA2 activities of all venoms were effectively inhibited by the small molecule inhibitor varespladib, suggesting its potential to be repurposed as a highly potent adjuvant therapeutic in snakebite envenoming.

Parasitoid wasp venoms degrade Drosophila imaginal discs for successful parasitism.

Parasitoid wasps, one of the most diverse and species-rich animal groups on Earth, produce venoms that manipulate host development and physiology to exploit resources. However, mechanisms of actions of these venoms remain poorly understood. Here, we discovered that the endoparasitoid wasp, Asobara japonica, induces apoptosis, autophagy, and mitotic arrest in the adult tissue precursors of its host Drosophila larvae. We termed this phenomenon imaginal disc degradation (IDD). A multi-omics approach facilitated identification of two venom proteins of A. japonica necessary for IDD, which is critical for parasitism success. Our study highlights a venom-mediated hijacking strategy of the parasitoid wasp that allows the host larvae to grow, but ultimately prevents their metamorphosis.

Recombinant venom proteins in insect seminal fluid reduce female lifespan

The emergence of insecticide resistance has increased the need for alternative pest management tools. Numerous genetic biocontrol approaches, which involve the release of genetically modified organisms to control pest populations, are in various stages of development to provide highly targeted pest control. However, all current mating-based genetic biocontrol technologies function by releasing engineered males which skew sex-ratios or reduce offspring viability in subsequent generations which leaves mated females to continue to cause harm (e.g. transmit disease). Here, we demonstrate intragenerational genetic biocontrol, wherein mating with engineered males reduces female lifespan. The toxic male technique (TMT) involves the heterologous expression of insecticidal proteins within the male reproductive tract that are transferred to females via mating. In this study, we demonstrate TMT in Drosophila melanogaster males, which reduce the median lifespan of mated females by 37 − 64% compared to controls mated to wild type males. Agent-based models of Aedes aegypti predict that TMT could reduce rates of blood feeding by a further 40 – 60% during release periods compared to leading biocontrol technologies like fsRIDL. TMT is a promising approach for combatting outbreaks of disease vectors and agricultural pests.

Metabolomics and proteomics: synergistic tools for understanding snake venom inhibition.

Snake envenomation presents a significant global health challenge, especially in rural areas of tropical and subtropical regions. Traditional antivenom therapies face limitations related to efficacy, availability, and specificity, prompting a need for novel approaches. Recent advancements in omics technologies, particularly metabolomics and proteomics, have enhanced our understanding of snake venom composition, toxicity, and potential therapeutic strategies. Metabolomics allows for the study of metabolic changes induced by venom, providing insights into disrupted pathways and possible inhibitors. Proteomics facilitates the identification and characterization of venom proteins, unveiling their interactions with therapeutic agents. Integrative databases such as the Snake Venom Database (SVDB) and STAB Profiles enhance this research by cataloging venom components and aiding in the analysis of venom-antivenom interactions. The combined application of metabolomics and proteomics has led to the identification of crucial metabolic pathways and protein targets essential for effective venom inhibition. This review explores current advances in these fields, emphasizing the role of omics in identifying novel inhibitors and developing next-generation antivenoms. The integrated approach of metabolomics and proteomics offers a comprehensive understanding of snake venom biology, paving the way for more effective and tailored therapeutic solutions for envenomation.

Proteogenomic approaches for snake venom protein-based drug development: current trends and challenges

Abstract Snake venom proteins have long been recognized for their therapeutic potential. Proteogenomic strategies, integrating transcriptomics and proteomics, have emerged as powerful tools for identifying and characterizing venom proteins for the development of novel therapeutic agents. Analytical techniques like mass spectrometry and next-generation sequencing enable comprehensive analysis, identifying key venom components and their variants. Recent studies unveil the diversity and complexity of snake venom, highlighting species-specific variations in toxin composition. Structural biology techniques, including x-ray crystallography and cryo-electron microscopy, provide insights into venom enzyme structures and mechanisms of action, facilitating drug design. Bioinformatics tools aid in data analysis and prediction of venom protein functions, enhancing drug discovery efforts. Despite advancements, challenges persist, including toxicity, formulation stability and clinical validation. This review describes the current as well as future options for research and development and emphasizes the critical role of proteogenomic techniques in developing snake venom protein-based drugs.

Prospect of using ethnobotanicals to manage snakebites in a cost-effective manner: validating Senegalia mellifera extract's inhibitory potential on Naja nigricincta nigricincta (zebra cobra) venom.

Despite Naja nigricincta nigricincta being responsible for most snake envenomation in remote Namibian regions, an effective intervention against its venom remains undiscovered. This study aimed to scientifically validate Namibian folklore claims about Senegalia mellifera extract's efficacy against snake envenomation. In vitro assays were conducted to assess the inhibitory potential of S. mellifera stem bark extract on snake venom phospholipase A2 (svPLA2) activity from N. n. nigricincta venom. Gas chromatography-mass spectrometry (GC-MS) and molecular docking predicted phytochemicals responsible for inhibitory effects on venom proteins. The svPLA2 activity assay demonstrated significant inhibitory potential of the extract, reducing enzyme activity from 100% to as low as 66.99%. GC-MS analysis indicated an abundant presence of terpenes having antisnake venom activity. Molecular docking identified phytochemicals of S. mellifera capable of neutralizing prevalent cobra toxins, that is, stigmasterol acetate, beta-Sitosterol acetate, vitamin E, kaur-15-ene, squalene and 4,6-Cholestadien-3beta-ol. This plant extract cannot be considered as a discrete treatment against venom. It may serve as a transient remedy to impede the toxic effects or supplement the action of antivenoms. Future research should be aimed at finding other plants with greater antivenom potential to increase the prospect of using ethnobotanicals to manage snakebites in a cost-effective manner.

Hydrophilic interaction chromatography coupled to high resolution mass spectrometry (HILIC-LC-HRMS): An approach to study natural peptides in Viperidae snake venom.

Although proteins in snake venoms have been extensively studied and characterized, low-mass molecules remain relatively unexplored, mainly due to their low abundance, secondary role in envenomation, and some analytical technique limitations. However, these small molecules can provide new important data related to venom toxins' molecular structure, functions, and evolutionary relationships. This research aimed to characterize molecules below 10 kDa in the venoms of snakes from the Viperidae families (Bothrops, Agkistrodon, and Bitis) and compare two chromatographic approaches: reverse-phase chromatography (RP), a classic technique, and hydrophilic interaction liquid chromatography (HILIC), an alternative technique, both coupled with high-resolution mass spectrometry (HRMS). The results showed that the separation of the HILIC column provided a more efficient evenly distributed ion profile than RP, contributing to a 25.6% increase in the sequences identified. Homologous sequences for Bradykinin-potentiating peptides (BPPs) and fragments of major venom proteins, possibly cryptids, were found. In addition, BPP 13a, peptides rich in histidine and glycine (pHpG), and spacer sequences were identified in all snakes analyzed, especially with HILIC separation, suggesting that these sequences may be conserved within Viperidae. These findings indicate that the use of the HILIC column, compared to RP, is a promising approach for characterizing peptides in snake venom obtained by the ultrafiltration process. It contributes to the study of these still poorly understood molecules and is also a good option for studying other complex protein/peptide mixtures.

Venomics AI: a computational exploration of global venoms for antibiotic discovery.

The relentless emergence of antibiotic-resistant pathogens, particularly Gram-negative bacteria, highlights the urgent need for novel therapeutic interventions. Drug-resistant infections account for approximately 5 million deaths annually, yet the antibiotic development pipeline has largely stagnated. Venoms, representing a remarkably diverse reservoir of bioactive molecules, remain an underexploited source of potential antimicrobials. Venom-derived peptides, in particular, hold promise for antibiotic discovery due to their evolutionary diversity and unique pharmacological profiles. In this study, we mined comprehensive global venomics datasets to identify new antimicrobial candidates. Using machine learning, we explored 16,123 venom proteins, generating 40,626,260 venom-encrypted peptides (VEPs). Using APEX, a deep learning model combining a peptide-sequence encoder with neural networks for antimicrobial activity prediction, we identified 386 VEPs structurally and functionally distinct from known antimicrobial peptides. Our analyses showed that these VEPs possess high net charge and elevated hydrophobicity, characteristics conducive to bacterial membrane disruption. Structural studies revealed considerable conformational flexibility, with many VEPs transitioning to α-helical conformations in membrane-mimicking environments, indicative of their antimicrobial potential. Of the 58 VEPs selected for experimental validation, 53 displayed potent antimicrobial activity. Mechanistic assays indicated that VEPs primarily exert their effects through bacterial membrane depolarization, mirroring AMP-like mechanisms. In vivo studies using a mouse model of Acinetobacter baumannii infection demonstrated that lead VEPs significantly reduced bacterial burdens without notable toxicity. This study highlights the value of venoms as a resource for new antibiotics. By integrating computational approaches and experimental validation, venom-derived peptides emerge as promising candidates to combat the global challenge of antibiotic resistance.

Integrative multi-omics analysis reveals the contribution of neoVTX genes to venom diversity of Synanceia verrucosa

BackgroundAnimal venom systems are considered as valuable model for investigating the molecular mechanisms underlying phenotypic evolution. Stonefish are the most venomous and dangerous fish because of severe human envenomation and occasionally fatalities, whereas the genomic background of their venom has not been fully explored compared with that in other venomous animals.ResultsIn this study, we followed modern venomic pipelines to decode the Synanceia verrucosa venom components. A catalog of 478 toxin genes was annotated based on our assembled chromosome-level genome. Integrative analysis of the high-quality genome, the transcriptome of the venom gland, and the proteome of crude venom revealed mechanisms underlying the venom complexity in S. verrucosa. Six tandem-duplicated neoVTX subunit genes were identified as the major source for the neoVTX protein production. Further isoform sequencing revealed massive alternative splicing events with a total of 411 isoforms demonstrated by the six genes, which further contributed to the venom diversity. We then characterized 12 dominantly expressed toxin genes in the venom gland, and 11 of which were evidenced to produce the venom protein components, with the neoVTX proteins as the most abundant. Other major venom proteins included a presumed CRVP, Kuntiz-type serine protease inhibitor, calglandulin protein, and hyaluronidase. Besides, a few of highly abundant non-toxin proteins were also characterized and they were hypothesized to function in housekeeping or hemostasis maintaining roles in the venom gland. Notably, gastrotropin like non-toxin proteins were the second highest abundant proteins in the venom, which have not been reported in other venomous animals and contribute to the unique venom properties of S. verrucosa.ConclusionsThe results identified the major venom composition of S. verrucosa, and highlighted the contribution of neoVTX genes to the diversity of venom composition through tandem-duplication and alternative splicing. The diverse neoVTX proteins in the venom as lethal particles are important for understanding the adaptive evolution of S. verrucosa. Further functional studies are encouraged to exploit the venom components of S. verrucosa for pharmaceutical innovation.

The Role of Snake Venom Proteins in Inducing Inflammation Post-Envenomation: An Overview on Mechanistic Insights and Treatment Strategies.

The intricate combination of organic and inorganic compounds found in snake venom includes proteins, peptides, lipids, carbohydrates, nucleotides, and metal ions. These components work together to immobilise and consume prey through processes such as paralysis and hypotension. Proteins, both enzymatic and non-enzymatic, form the primary components of the venom. Based on the effects they produce, venom can be classified as neurotoxic, hemotoxic, and cytotoxic. Studies have shown that, after envenomation, proteins in snake venom also contribute significantly to the induction of inflammatory responses which can either have systemic or localized consequences. This review delves into the mechanisms by which snake venom proteins trigger inflammatory responses, focusing on key families such as phospholipase A2, metalloproteinases, serine proteases, C-type lectins, cysteine-rich secretory proteins, and L-amino acid oxidase. In addition, the role of venom proteins in activating various inflammatory pathways, including the complement system, inflammasomes, and sterile inflammation are also summarized. The available therapeutic options are examined, with a focus on antivenom therapy and its side effects. In general, this review offers a comprehensive understanding of the inflammatory mechanisms that are triggered by snake venom proteins and the side effects of antivenom treatment. All these emphasize the need for effective strategies to mitigate these detrimental effects.

Insecticidal activities of three recombinant venom proteins of the predatory stink bug, Arma custos.

Widespread resistance of insect pests to insecticides and transgenic crops in the field is a significant challenge for sustainable agriculture, and calls for the development of novel alternative strategies to control insect pests. One potential resource for the discovery of novel insecticidal molecules is natural toxins, particularly those derived from the venoms of insect predators. In this study, we identified three insecticidal proteinaceous toxins from the venom glands (VGs) of the predatory stink bug, Arma custos (Hemiptera: Asopinae). Transcriptomic analysis of A. custos VGs revealed 151 potentially secreted VG-rich venom proteins. Three VG-rich venom proteins (designated AcVP1 ~ 3) were produced by overexpression in Escherichia coli. Injection of the recombinant proteins into tobacco cutworm (Spodoptera litura) larvae showed that all of the three recombinant proteins caused paralysis, liquefaction and death. Injection of recombinant proteins into rice brown planthopper (Nilaparvata lugens) nymphs showed higher insecticidal activities, among which a trypsin (AcVP2) caused 100% mortality postinjection at 1.27 pmol mg-1 body weight. A natural toolkit for the discovery of insecticidal toxins from predatory insects has been revealed by the present study. © 2024 Society of Chemical Industry.

Improving parasitism success of a weakly virulent parasitoid strain.

Endoparasitoids possess a whole set of virulence factors to counter the immune response of their host, among which can be found venom, endosymbiotic viruses and ovarian proteins. Depending on the species, some factors are expected to be less necessary than others. Notably, venom is reported as inessential in some parasitoids bearing viruses. We investigated the virulence factors of Cotesia typhae, a gregarious endoparasitoid of Sesamia nonagrioides, using highly and poorly virulent strains (CtV+and CtV-). We tested if virulence of CtV- toward a reference strain could be improved by superparasitism (two females per host) or by injection of CtV+virulence factors (venom and/or ovarian fluid). The parasitism success of CtV- could be increased by superparasitism with one female CtV+confirming the importance of the virulence factors. Superparasitism with at least one CtV+female decreased each female's reproductive success and increased the number of non-viable offspring revealing larval competition. Parasitism order impacted the offspring proportion of each strain in the progeny, suggesting adaptation of egg-laying behavior in response to an already parasitized host. Injection of CtV+venom or ovarian fluid alone maintained CtV- success unchanged, but their combination increased it to the one of CtV+. Altogether, this study hints towards larval competition, potentially specific to CtV+strain, and suggests that C. typhae females can adapt their oviposition behavior based on the parasitic status of their host. It also demonstrates that, like several other Cotesia species, C. typhae resorts to a combination of venom and ovarian fluid to overcome the host's immune responses.

Unraveling the venom constituents of the endoparasitoid Aphidius gifuensis with an emphasis on the discovery of a novel insecticidal peptide.

Venom serves as a pivotal parasitic factor employed by parasitoid wasps to manipulate their hosts, creating a favorable environment for the successful growth of their progeny, and ultimately kill the host. The bioactive molecules within parasitoid venoms exhibit insecticidal activities with promising prospects for agricultural applications. However, knowledge regarding the venom components of parasitoids and the discovery of functional biomolecules from them remains limited. In this study, 30 venom proteins were identified from the endoparasitoid Aphidius gifuensis through the application of a transcriptomic approach. These proteins were categorized into five groups: hydrolase, molecular chaperone, transferase, other functional protein, and hypothetical protein with unknown function. Particularly noteworthy is the abundant expression of the peptide Vn1 in the venom apparatus of A. gifuensis, indicating its pivotal role in venom activity. Consequently, Vn1 was chosen for further functional analysis, exhibiting insecticidal activity against Tenebrio molitor pupae. Further assessment for revealing its mode of action disclosed that Vn1 impacts genes related to immune response, environmental information processing, metabolism, and response to external stimuli in T. molitor, suggesting its involvement in the intricate parasitoid wasp-host interaction. The findings of this study significantly contribute to our knowledge of the composition and functionality of A. gifuensis venom, establishing a foundation for further investigation into the biological roles of the identified venom constituents. The insecticidal Vn1 isolated from the venom of this parasitoid represents a valuable resource for the development of innovative biocontrol agents with potential applications in agriculture. © 2024 Society of Chemical Industry.

Top-down proteomics of venoms from five Micrurus species from Costa Rica: Comparative composition of phospholipase A2-rich vs three-finger toxin-rich phenotypes

Coralsnakes of the genus Micrurus include more than 80 species distributed in the American continent. They produce potent neurotoxic venoms acting at the neuromuscular junction and potentially leading to respiratory paralysis and death. The vast majority of proteins in coralsnake venoms belong to the three-finger toxin (3FTx) and the group I phospholipase A2 (PLA2) families. Previous studies using 'bottom-up' proteomic strategies have revealed a compositional dichotomy of toxin expression by which different Micrurus species display a predominance of either 3FTx or PLA2 proteins in their venoms, possibly linked to the phylogeographic structure of the genus radiation. 'Top-down' proteomics (TDP) allows the direct analysis of intact proteins in a high resolution mass spectrometer, circumventing the limitations of the 'peptide-to-protein inference problem' inherent to the bottom-up approach. Here, we analyzed the venoms of five out of the six Micrurus species that inhabit Costa Rica, by using a TDP approach. Results unveil venom proteoforms that are shared between these species, and provide additional insights into the variable compositional complexity of these venoms and relationships to their 3FTx/PLA2 dichotomy.

ScorpDb: A Novel Open-Access Database for Integrative Scorpion Toxinology.

Scorpion stings are a significant public health concern globally, particularly in tropical and subtropical regions. Scorpion venoms contain a diverse array of bioactive peptides, and different scorpion species around the world typically exhibit varying venom profiles, resulting in a wide range of envenomation symptoms. Despite their harmful effects, scorpion venom peptides hold immense potential for drug development due to their unique characteristics. Therefore, the establishment of a comprehensive database that catalogs scorpions along with their known venom peptides and proteins is imperative in furthering research efforts in this research area. We hereby present ScorpDb, a novel database that offers convenient access to data related to different scorpion species, the peptides and proteins found in their venoms, and the symptoms they can cause. To this end, the ScorpDb database has been primarily advanced to accommodate data on the Iranian scorpion fauna. From there, we propose future community efforts to include a larger diversity of scorpions and scorpion venom components. ScorpDb holds the promise to become a valuable resource for different professionals from a variety of research fields, like toxinologists, arachnologists, and pharmacologists. The database is available at https://www.scorpdb.com/.

First transcriptome analysis of the venom glands of the scorpion Hottentotta zagrosensis (Scorpions: Buthidae) with focus on venom lipolysis activating peptides.

Scorpion venom is a rich source of biological active peptides and proteins. Transcriptome analysis of the venom gland provides detailed insights about peptide and protein venom components. Following the transcriptome analysis of different species in our previous studies, our research team has focused on the Hottentotta zagrosensis as one of the endemic scorpions of Iran to obtain information about its venom proteins, in order to develop biological research focusing on medicinal applications of scorpion venom components and antivenom production. To gain insights into the protein composition of this scorpion venom, we performed transcriptomic analysis. Transcriptomic analysis of the venom gland of H. zagrosensis, prepared from the Khuzestan province, was performed through Illumina paired-end sequencing (RNA-Seq), Trinity de novo assembly, CD-Hit-EST clustering, and annotation of identified primary structures using bioinformatics approaches. Transcriptome analysis showed the presence of 96.4% of complete arthropod BUSCOs, indicating a high-quality assembly. From total of 45,795,108 paired-end 150 bp trimmed reads, the clustering step resulted in the generation of 101,180 de novo assembled transcripts with N50 size of 1,149 bp. 96,071 Unigenes and 131,235 transcripts had a significant similarity (E-value 1e-3) with known proteins from UniProt, Swissprot, Animal toxin annotation project, and the Pfam database. The results were validated using InterProScan. These mainly correspond to ion channel inhibitors, metalloproteinases, neurotoxins, protease inhibitors, protease activators, Cysteine-rich secretory proteins, phospholipase A enzymes, antimicrobial peptides, growth factors, lipolysis-activating peptides, hyaluronidase, and, phospholipase D. Our venom gland transcriptomic approach identified several biologically active peptides including five LVP1-alpha and LVP1-beta isoforms, which we named HzLVP1_alpha1, HzLVP1_alpha2, HzLVP1_alpha3, HzLVP1_beta1, and HzLVP1_beta and have extremely characterized here. Except for HzLVP1_beta1, all other identified LVP1s are predicted to be stable proteins (instability index <40). Moreover, all isoform of LVP1s alpha and beta subunits are thermostable, with the most stability for HzLVP1_alpha2 (aliphatic index = 71.38). HzLVP1_alpha2 has also the highest half-life. Three-dimensional structure of all identified proteins compacts with three disulfide bridges. The extra cysteine residue may allow the proteins to form a hetero- or homodimer. LVP1 subunits of H. zagrosensis potentially interact with adipose triglyceride lipase (ATGL) and hormone-sensitive lipase (HSL), two key enzymes in regulation of lipolysis in adipocytes, suggesting pharmacological properties of these identified proteins.

Snake venom weakens neurovascular integrity and promotes vulnerability to neuroinflammation

Snake envenomation poses significant medical challenges, particularly in subtropical and tropical regions, with long-term impacts on neurovascular integrity and neuroinflammation remaining underexplored. This study investigates the effects of venom from four species of venomous snakes in southern China-Zhoushan Cobra (Naja atra, NA), Many-banded Krait (Bungarus multicinctus, BM), Five-paced Pit Viper (Deinagkistrodon acutus, DA), and Chinese Moccasin (Protobothrops mucrosquamatus, PM) - on the blood-brain barrier (BBB) and chronic neuroinflammation. Using mass spectrometry, we analyzed venom protein compositions, while cytotoxic effects on mouse brain endothelial cells (bEND.3) were evaluated to determine IC50 values. In vitro BBB models and in vivo experiments in C57BL/6J mice revealed that NA venom, in particular, significantly compromised BBB integrity without inducing large-scale apoptosis, leading to persistent BBB disruption characterized by increased permeability and selective degradation of extracellular matrix and tight junction proteins. Moreover, to simulate secondary infections that often occur following snakebites, we combined venom exposure with lipopolysaccharide (LPS) treatment, which exacerbated neuroinflammatory responses by intensifying microglial activation and promoting a pro-inflammatory phenotype. These findings highlight the role of snake venom in compromising neurovascular integrity and promoting vulnerability to chronic neuroinflammation, emphasizing the need for further research into venom-induced neuroinflammatory pathways and their potential as therapeutic targets.

Exploring secretory signal sequences useful in excreting recombinant proteins in Beauveria bassiana as biocontrol fungus.

Entomopathogenic fungi excrete a group of proteins to assimilate nutrients and defeat the host immune defense. Functional secretory signal sequences are needed for efficient secretion of the virulence-related proteins in recombinant strain. In this study, secretome analysis was used to explore the secreted proteins of Beauveria bassiana. Enrichment analysis indicated that B. bassiana secretome was mainly associated with metabolism of glucoside, polysaccharide, extracellular ester compound, and so on. In addition, proteins associated with biogenesis of cellular components were also enriched, including those involved in biogenesis of cell wall and vacuole. Then, four secretory signal sequences were functionally verified with green fluorescent protein as reporter. Finally, a signal sequence was used to excrete three insect venom protein serpins in B. bassiana, in which over-expression of serpin 8 gene resulted in a significant increase in fungal virulence. This study highlights that functional secretory signal sequences are potential molecular elements useful in excretion of virulence-related proteins in insect pathogenic fungi.

Regulation of melanization in aphids by parasitoid wasp venom proteins enhances mummification.

Interactions between parasitic insects and their hosts demonstrate the complexity of evolutionary processes. Specifically, the parasitoid wasp Aphidius ervi manipulates its host, the pea aphid Acyrthosiphon pisum, through strategic venom injection to enhance mummification. This study explores how this venom affects the aphid's immune system, particularly targeting the activity of the phenoloxidase (PO) enzyme. Following the injection of venom from A. ervi, significant changes were observed in the expression of immune-related genes in A. pisum, especially notable expression changes of ApPPOs and a reduction of PO activity. Multi-omics sequencing identified 74 potential venom proteins in the venom gland of A. ervi, including serine protease homolog 1 (AeSPH1) and serine protease inhibitor (AeSPN1), hypothesized to regulate PO activity. The injection of recombinant protein AeSPH1 and AeSPN1 into the A. pisum hemocoel selectively reduced the expression of ApPPO1, without affecting ApPPO2, and effectively suppressed melanization. Moreover, RNAi targeting AeSPH1 significantly reduced the mummification rate in A. pisum population parasitized by A. ervi. Our findings clarify the complex biochemical mechanisms underlying host-wasp interactions and highlight potential avenues for developing targeted biological control strategies. © 2024 Society of Chemical Industry.