The Peptide Venom Composition of the Fierce Stinging Ant Tetraponera aethiops (Formicidae: Pseudomyrmecinae)

In the mutualisms involving certain pseudomyrmicine ants and different myrmecophytes (i.e., plants sheltering colonies of specialized “plant-ant” species in hollow structures), the ant venom contributes to the host plant biotic defenses by inducing the rapid paralysis of defoliating insects and causing intense pain to browsing mammals. Using integrated transcriptomic and proteomic approaches, we identified the venom peptidome of the plant-ant Tetraponera aethiops (Pseudomyrmecinae). The transcriptomic analysis of its venom glands revealed that 40% of the expressed contigs encoded only seven peptide precursors related to the ant venom peptides from the A-superfamily. Among the 12 peptide masses detected by liquid chromatography-mass spectrometry (LC–MS), nine mature peptide sequences were characterized and confirmed through proteomic analysis. These venom peptides, called pseudomyrmecitoxins (PSDTX), share amino acid sequence identities with myrmeciitoxins known for their dual offensive and defensive functions on both insects and mammals. Furthermore, we demonstrated through reduction/alkylation of the crude venom that four PSDTXs were homo- and heterodimeric. Thus, we provide the first insights into the defensive venom composition of the ant genus Tetraponera indicative of a streamlined peptidome.

Download Full-text

Proteotranscriptomic Insights into the Venom Composition of the Wolf Spider Lycosa tarantula

Toxins ◽

10.3390/toxins12080501 ◽

2020 ◽

Vol 12 (8) ◽

pp. 501

Author(s):

Dominique Koua ◽

Rosanna Mary ◽

Anicet Ebou ◽

Celia Barrachina ◽

Khadija El Koulali ◽

...

Keyword(s):

Prey Capture ◽

Wolf Spider ◽

Secretory Proteins ◽

Crude Venom ◽

Wolf Spiders ◽

Original Source ◽

Venom Composition ◽

Venom Glands ◽

Ambush Predators ◽

Venom Proteins

Spider venoms represent an original source of novel compounds with therapeutic and agrochemical potential. Whereas most of the research efforts have focused on large mygalomorph spiders, araneomorph spiders are equally promising but require more sensitive and sophisticated approaches given their limited size and reduced venom yield. Belonging to the latter group, the genus Lycosa (“wolf spiders”) contains many species widely distributed throughout the world. These spiders are ambush predators that do not build webs but instead rely strongly on their venom for prey capture. Lycosa tarantula is one of the largest species of wolf spider, but its venom composition is unknown. Using a combination of RNA sequencing of the venom glands and venom proteomics, we provide the first overview of the peptides and proteins produced by this iconic Mediterranean spider. Beside the typical small disulfide rich neurotoxins, several families of proteins were also identified, including cysteine-rich secretory proteins (CRISP) and Hyaluronidases. Proteomic analysis of the electrically stimulated venom validated 30 of these transcriptomic sequences, including nine putative neurotoxins and eight venom proteins. Interestingly, LC-MS venom profiles of manual versus electric stimulation, as well as female versus male, showed some marked differences in mass distribution. Finally, we also present some preliminary data on the biological activity of L. tarantula crude venom.

Download Full-text

TOXIC AND IMMUNE ALLERGIC RESPONSES OF ANT VENOM TOXINS: A REVIEW

International Journal of Pharmacy and Pharmaceutical Sciences ◽

10.22159/ijpps.2021v13i11.42227 ◽

2021 ◽

pp. 1-7

Author(s):

SIMRAN SHARMA ◽

RAVI KANT UPADHYAY!

Keyword(s):

Immune Responses ◽

Biological Effects ◽

Therapeutic Agents ◽

Microbial Pathogens ◽

Intense Pain ◽

Venom Toxins ◽

Potential Source ◽

Ant Venom ◽

Peptide Toxins ◽

Promising Source

Present review article explains ant venom components and its allergic and biological effects in man and animals. Red ants or small fire ants secrete and inject venom very swiftly to defend their nest against predators, microbial pathogens, and competitors and to hunt the prey. Ant venom is a mixture of various organic compounds, including peptides, enzymes, and polypeptide toxins. It is highly toxic, allergic, invasive and venomous. It imposes sever paralytic, cytolytic, haemolytic, allergenic, pro-inflammatory, insecticidal, antimicrobial, and pain-producing pharmacologic activities after infliction. Victims show red ring-shaped allergic sign with regional swelling marked with intense pain. Ant venom also contains several hydrolases, oxidoreductases, proteases, Kunitz-like polypeptides, and inhibitor cysteine knot (ICK)-like (knottin) neurotoxins and insect defensins. Ant venom toxins/proteins generate allergic immune responses and employ eosinophils and produce Th2 cytokines, response. These compounds from ant venom could be used as a potential source of new anticonvulsants molecules. Ant venoms contain many small, linear peptides, an untapped source of bioactive peptide toxins. The remarkable insecticidal activity of ant venom could be used as a promising source of additional bio-insecticides and therapeutic agents.

Download Full-text

Novel Pharmacological Properties of Dinoponera quadriceps Giant Ant Venom

Natural Product Communications ◽

10.1177/1934578x1501000930 ◽

2015 ◽

Vol 10 (9) ◽

pp. 1934578X1501000 ◽

Cited By ~ 1

Author(s):

Juliana da Costa Madeira ◽

Yves Patric Quinet ◽

Dayanne Terra Tenório Nonato ◽

Paloma Leão Sousa ◽

Edna Maria Camelo Chaves ◽

...

Keyword(s):

Platelet Aggregation ◽

Anticoagulant Activity ◽

Adenosine Diphosphate ◽

South American ◽

Crude Venom ◽

Leucocyte Migration ◽

Ant Venom ◽

Hymenopteran Species ◽

Anti Inflammatory

The South American giant ant, Dinoponera quadriceps (Hymenoptera, Formicidae, Ponerinae), produces proteinaceous venom that has antinociceptive, neuroprotective and antimicrobial effects, thereby supporting the popular use of these ants to treat asthma, rheumatism, earache and back pain. Anticoagulant activity is another biological property that has been shown for the venom of other hymenopteran species, like wasps. The aim of this study was to assess the anti-inflammatory, anticoagulant and antiplatelet properties of D. quadriceps venom (DqV). DqV anti-inflammatory activity was assessed by intravenous administration in Swiss mice in the models of paw edema and peritonitis. In vitro, DqV was assessed in coagulation (activated partial thromboplastin time) and platelet aggregation tests. DqV inhibited (27–33%) the edema elicited by carrageenan and the leucocyte migration (43%) elicited by zymosan. DqV decreased by 57% and 42%, respectively, the content of malondialdehyde and nitrite in the peritoneal fluid. DqV prolonged (1.8x) the clotting time and decreased (27%) the platelet aggregation induced by adenosine diphosphate. The crude venom of D. quadriceps presents an anti-inflammatory effect in mice and in vitro anticoagulant and antiplatelet effects.

Download Full-text

An Integrated Proteomic and Transcriptomic Analysis Reveals the Venom Complexity of the Bullet Ant Paraponera clavata

Toxins ◽

10.3390/toxins12050324 ◽

2020 ◽

Vol 12 (5) ◽

pp. 324

Author(s):

Samira R. Aili ◽

Axel Touchard ◽

Regan Hayward ◽

Samuel D. Robinson ◽

Sandy S. Pineda ◽

...

Keyword(s):

Mass Spectrometry ◽

Venom Gland ◽

Sequence Database ◽

Liquid Chromatography Mass Spectrometry ◽

Ant Venom ◽

Proteomic Investigation ◽

Paraponera Clavata ◽

And Function ◽

Venom Proteins ◽

Proteins And Peptides

A critical hurdle in ant venom proteomic investigations is the lack of databases to comprehensively and specifically identify the sequence and function of venom proteins and peptides. To resolve this, we used venom gland transcriptomics to generate a sequence database that was used to assign the tandem mass spectrometry (MS) fragmentation spectra of venom peptides and proteins to specific transcripts. This was performed alongside a shotgun liquid chromatography–mass spectrometry (LC-MS/MS) analysis of the venom to confirm that these assigned transcripts were expressed as proteins. Through the combined transcriptomic and proteomic investigation of Paraponera clavata venom, we identified four times the number of proteins previously identified using 2D-PAGE alone. In addition to this, by mining the transcriptomic data, we identified several novel peptide sequences for future pharmacological investigations, some of which conform with inhibitor cysteine knot motifs. These types of peptides have the potential to be developed into pharmaceutical or bioinsecticide peptides.

Download Full-text

Electric Blue: Molecular Evolution of Three-Finger Toxins in the Long-Glanded Coral Snake Species Calliophis bivirgatus

Toxins ◽

10.3390/toxins13020124 ◽

2021 ◽

Vol 13 (2) ◽

pp. 124

Author(s):

Daniel Dashevsky ◽

Darin Rokyta ◽

Nathaniel Frank ◽

Amanda Nouwens ◽

Bryan G. Fry

Keyword(s):

Molecular Characteristics ◽

Substantial Portion ◽

Coral Snake ◽

Spastic Paralysis ◽

Voltage Gated ◽

Venom Composition ◽

The Family ◽

Basal Branch ◽

Venom Glands ◽

Nav Channels

The genus Calliophis is the most basal branch of the family Elapidae and several species in it have developed highly elongated venom glands. Recent research has shown that C. bivirgatus has evolved a seemingly unique toxin (calliotoxin) that produces spastic paralysis in their prey by acting on the voltage-gated sodium (NaV) channels. We assembled a transcriptome from C. bivirgatus to investigate the molecular characteristics of these toxins and the venom as a whole. We find strong confirmation that this genus produces the classic elapid eight-cysteine three-finger toxins, that δδ-elapitoxins (toxins that resemble calliotoxin) are responsible for a substantial portion of the venom composition, and that these toxins form a distinct clade within a larger, more diverse clade of C. bivirgatus three-finger toxins. This broader clade of C. bivirgatus toxins also contains the previously named maticotoxins and is somewhat closely related to cytotoxins from other elapids. However, the toxins from this clade that have been characterized are not themselves cytotoxic. No other toxins show clear relationships to toxins of known function from other species.

Download Full-text

Proteo-Transcriptomic Characterization of Sirex nitobei (Hymenoptera: Siricidae) Venom

Toxins ◽

10.3390/toxins13080562 ◽

2021 ◽

Vol 13 (8) ◽

pp. 562

Author(s):

Chenglong Gao ◽

Lili Ren ◽

Ming Wang ◽

Zhengtong Wang ◽

Ningning Fu ◽

...

Keyword(s):

Coniferous Forest ◽

Venom Gland ◽

Crude Venom ◽

Symbiotic Fungus ◽

Host Trees ◽

Liquid Chromatography Tandem Mass ◽

Venom Glands ◽

Encoding Protein ◽

First Time

The wood-boring woodwasp Sirex nitobei is a native pest in Asia, infecting and weakening the host trees in numerous ecological and commercial coniferous forest plantations. In China, hosts of S. nitobei are diverse, so the pest has spread to several provinces of China, resulting in considerable economic and ecological damage. During female oviposition, S. nitobei venom along with arthrospores of the symbiotic fungus Amylostereum areolatum or A. chaetica is injected into host trees, and the combination of these two biological factors causes the death of xylem host trees. The presence of venom alone causes only the yellowing and wilting of needles. In this study, we constructed the venom gland transcriptome of S. nitobei for the first time and a total of 15,036 unigenes were acquired. From the unigenes, 11,560 ORFs were identified and 537 encoding protein sequences with signal peptides at the N-terminus. Then, we used the venomics approach to characterize the venom composition of female S. nitobei and predicted 1095 proteins by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. We focused on seven proteins that were both highly expressed in the venom gland transcriptome and predicted in the crude venom proteome. These seven proteins are laccase-2, laccase-3, a protein belonging to the Kazal family, chitooligosaccharidolytic β-N-acetylglucosaminidase, beta-galactosidase, icarapin-like protein, and waprin-Thr1-like protein. Using quantitative real-time PCR (qRT-PCR), we also proved that the genes related to these seven proteins are specifically expressed in the venom glands. Finally, we revealed the functional role of S. nitobei venom in the physiological response of host trees. It can not only promote the colonization of symbiotic fungus but contribute to the development of eggs and larvae. This study provides a deeper understanding of the molecular mechanism of the woodwasp–pine interaction.

Download Full-text

New Insectotoxin from Tibellus Oblongus Spider Venom Presents Novel Adaptation of ICK Fold

Toxins ◽

10.3390/toxins13010029 ◽

2021 ◽

Vol 13 (1) ◽

pp. 29

Author(s):

Yuliya Korolkova ◽

Ekaterina Maleeva ◽

Alexander Mikov ◽

Anna Lobas ◽

Elizaveta Solovyeva ◽

...

Keyword(s):

Disulfide Bonds ◽

Water Solution ◽

Conformational Stability ◽

Evolutionary Adaptation ◽

Distinctive Features ◽

E Coli ◽

Cdna Analysis ◽

Venom Composition ◽

Venom Glands ◽

Active Predator

The Tibellus oblongus spider is an active predator that does not spin webs and remains poorly investigated in terms of venom composition. Here, we present a new toxin, named Tbo-IT2, predicted by cDNA analysis of venom glands transcriptome. The presence of Tbo-IT2 in the venom was confirmed by proteomic analyses using the LC-MS and MS/MS techniques. The distinctive features of Tbo-IT2 are the low similarity of primary structure with known animal toxins and the unusual motif of 10 cysteine residues distribution. Recombinant Tbo-IT2 (rTbo-IT2), produced in E. coli using the thioredoxin fusion protein strategy, was structurally and functionally studied. rTbo-IT2 showed insecticidal activity on larvae of the housefly Musca domestica (LD100 200 μg/g) and no activity on the panel of expressed neuronal receptors and ion channels. The spatial structure of the peptide was determined in a water solution by NMR spectroscopy. The Tbo-IT2 structure is a new example of evolutionary adaptation of a well-known inhibitor cystine knot (ICK) fold to 5 disulfide bonds configuration, which determines additional conformational stability and gives opportunities for insectotoxicity and probably some other interesting features.

Download Full-text

Composition and Acute Inflammatory Response from Tetraponera rufonigra Venom on RAW 264.7 Macrophage Cells

Toxins ◽

10.3390/toxins13040257 ◽

2021 ◽

Vol 13 (4) ◽

pp. 257

Author(s):

Suwatjanee Naephrai ◽

Supakit Khacha-ananda ◽

Pornsiri Pitchakarn ◽

Churdsak Jaikang

Keyword(s):

Protein Composition ◽

Raw 264.7 ◽

Dependent Manner ◽

Crude Venom ◽

Venom Protein ◽

Macrophage Cells ◽

Raw 264.7 Macrophage ◽

Ant Venom ◽

Raw 264.7 Macrophage Cells ◽

Cox 2

Tetraponera rufonigra (Arboreal Bicoloured Ant) venom induces pain, inflammation, and anaphylaxis in people and has an increased incident in Southeast Asia regions. The bioactive components and mechanism of action of the ant venom are still limited. The aim of this research was to identify the protein composition and inflammatory process of the ant venom by using RAW 264.7 macrophage cells. The major venom proteins are composed of 5’ nucleotidase, prolyl endopeptidase-like, aminopeptidase N, trypsin-3, venom protein, and phospholipase A2 (PLA2). The venom showed PLA2 activity and represented 0.46 μg of PLA2 bee venom equivalent/μg crude venom protein. The venom induced cytotoxic in a dose- and time-dependent manner with IC20 approximately at 4.01 µg/mL. The increased levels of COX-2 and PGE2 were observed after 1 h of treatment correlating with an upregulation of COX-2 expression. Moreover, the level of mPGES-1 expression was obviously increased after 12 h of venom induction. Hence, our results suggested that the induction of COX-2/mPGEs-1 pathway could be a direct pathway for the ant venom-induced inflammation.

Download Full-text

Bottom-Up Proteomic Analysis of Polypeptide Venom Components of the Giant Ant Dinoponera Quadriceps

Toxins ◽

10.3390/toxins11080448 ◽

2019 ◽

Vol 11 (8) ◽

pp. 448 ◽

Cited By ~ 7

Author(s):

Douglas Oscar Ceolin Mariano ◽

Úrsula Castro de Oliveira ◽

André Junqueira Zaharenko ◽

Daniel Carvalho Pimenta ◽

Gandhi Rádis-Baptista ◽

...

Keyword(s):

Biological Effects ◽

Glucose Dehydrogenase ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Crude Venom ◽

Venom Protein ◽

Bottom Up ◽

Venom Composition ◽

Proteomics Approach ◽

Analytical Approaches

Ant species have specialized venom systems developed to sting and inoculate a biological cocktail of organic compounds, including peptide and polypeptide toxins, for the purpose of predation and defense. The genus Dinoponera comprises predatory giant ants that inoculate venom capable of causing long-lasting local pain, involuntary shaking, lymphadenopathy, and cardiac arrhythmias, among other symptoms. To deepen our knowledge about venom composition with regard to protein toxins and their roles in the chemical–ecological relationship and human health, we performed a bottom-up proteomics analysis of the crude venom of the giant ant D. quadriceps, popularly known as the “false” tocandiras. For this purpose, we used two different analytical approaches: (i) gel-based proteomics approach, wherein the crude venom was resolved by denaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and all protein bands were excised for analysis; (ii) solution-based proteomics approach, wherein the crude venom protein components were directly fragmented into tryptic peptides in solution for analysis. The proteomic data that resulted from these two methodologies were compared against a previously annotated transcriptomic database of D. quadriceps, and subsequently, a homology search was performed for all identified transcript products. The gel-based proteomics approach unequivocally identified nine toxins of high molecular mass in the venom, as for example, enzymes [hyaluronidase, phospholipase A1, dipeptidyl peptidase and glucose dehydrogenase/flavin adenine dinucleotide (FAD) quinone] and diverse venom allergens (homologous of the red fire ant Selenopsis invicta) and venom-related proteins (major royal jelly-like). Moreover, the solution-based proteomics revealed and confirmed the presence of several hydrolases, oxidoreductases, proteases, Kunitz-like polypeptides, and the less abundant inhibitor cysteine knot (ICK)-like (knottin) neurotoxins and insect defensin. Our results showed that the major components of the D. quadriceps venom are toxins that are highly likely to damage cell membranes and tissue, to cause neurotoxicity, and to induce allergic reactions, thus, expanding the knowledge about D. quadriceps venom composition and its potential biological effects on prey and victims.

Download Full-text