Spatial Venomics─Cobra Venom System Reveals Spatial Differentiation of Snake Toxins by Mass Spectrometry Imaging

Hempel, Benjamin-Florian; Damm, Maik; Petras, Daniel; Kazandjian, Taline D.; Szentiks, Claudia A.; Fritsch, Guido; Nebrich, Grit; Casewell, Nicholas R.; Klein, Oliver

doi:10.1021/acs.jproteome.2c00424

Cited by 10 publications

(6 citation statements)

References 52 publications

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“…These proteins have been found in submandibular glands of other nonvenomous shrews such as Suncus murinus and Crocidura horsfieldi [ 63 , 64 ], and in rodents (rats, mice, hamsters, and voles) [ 64 – 66 ]. NGFs are also important components of snake venoms [ 67 ]. NGF expression is increased in mice submandibular salivary glands in response to chronic stress.…”

Section: Discussionmentioning

confidence: 99%

Proteins from shrews’ venom glands play a role in gland functioning and venom production

Kowalski,

Marciniak,

Nekaris

et al. 2024

Zoological Lett

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Venom production has evolved independently many times in the animal kingdom, although it is rare among mammals. Venomous shrews produce venom in their submandibular salivary glands and use it for food acquisition. Only a few toxins have been identified in shrew venoms thus far, and their modes of action require investigation. The biological and molecular processes relating to venom production and gland functioning also remain unknown. To address this gap, we investigated protein content in extracts from venom glands of two shrew species, Neomys fodiens and Sorex araneus, and interpreted their biological functions. Applying a proteomic approach coupled with Gene Ontology enrichment analysis, we identified 313 and 187 putative proteins in venom glands of N. fodiens and S. araneus, respectively. A search of the UniProt database revealed that most of the proteins found in both shrew species were involved in metabolic processes and stress response, while GO enrichment analysis revealed more stress-related proteins in the glands of S. araneus. Molecules that regulate molecule synthesis, cell cycles, and cell divisions are necessary to enable venom regeneration and ensure its effectiveness in predation and food hoarding. The presence of proteins involved in stress response may be the result of shrews’ high metabolic rate and the costs of venom replenishment. Some proteins are likely to promote toxin spreading during envenomation and, due to their proteolytic action, reinforce venom toxicity. Finally, finding numerous proteins involved in immune response suggests a potential role of shrew venom gland secretions in protection against pathogens. These findings open up new perspectives for studying biological functions of molecules from shrew venom glands and extend our knowledge on the functioning of eulipotyphlan venom systems. Because the majority of existing and putative venomous mammals use oral venom systems to inject venom into target species, the methods presented here provide a promising avenue for confirming or discovering new taxa of venomous mammals.

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Section: Discussionmentioning

confidence: 99%

Proteins from shrews’ venom glands play a role in gland functioning and venom production

Kowalski,

Marciniak,

Nekaris

et al. 2024

Zoological Lett

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“…The used bottom-up protocol is adapted from published protocols. , In short, 1 mg lyophilized venom was fractionated by HPLC, observed at 214 nm. Collected peaks were submitted to SDS-PAGE profiling and in-gel tryptic digestion, followed by LC–MS/MS measurements.…”

Section: Methodsmentioning

confidence: 99%

“…The used top-down protocol is adapted from published protocols. , In short, 100 μg lyophilized venom was measured reduced and nonreduced. Ten μL of each sample was injected into an Q Exactive HF mass spectrometer (Thermo, Bremen, Germany) via a Vanquish ultrahigh performance liquid chromatography (UHPLC) system (Agilent Technologies, Waldbronn, Germany) using a reversed-phase Supelco Discovery BIO wide C18 (2.0 × 150 mm; 3 μm particle size; 300 Å pore size) column thermostated at 30 °C.…”

Section: Methodsmentioning

confidence: 99%

Venomics and Peptidomics of Palearctic Vipers: A Clade-Wide Analysis of Seven Taxa of the Genera Vipera, Montivipera, Macrovipera, and Daboia across Türkiye

Damm,

Karış,

Petras

et al. 2024

J. Proteome Res.

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Snake venom variations are a crucial factor to understand the consequences of snakebite envenoming worldwide, and therefore it is important to know about toxin composition alterations between taxa. Palearctic vipers of the genera Vipera, Montivipera, Macrovipera, and Daboia have high medical impacts across the Old World. One hotspot for their occurrence and diversity is Turkiye, located on the border between continents, but many of their venoms remain still understudied. Here, we present the venom compositions of seven Turkish viper taxa. By complementary mass spectrometry-based bottom-up and topdown workflows, the venom profiles were investigated on proteomics and peptidomics level. This study includes the first venom descriptions of Vipera berus barani, Vipera darevskii, Montivipera bulgardaghica albizona, and Montivipera xanthina, as well as the first snake venomics profiles of Turkish Macrovipera lebetinus obtusa, and Daboia palaestinae, including an in-depth reanalysis of M. bulgardaghica bulgardaghica venom. Additionally, we identified the modular consensus sequence pEXW(PZ) 1−2 P(EI)/(KV)PPLE for bradykinin-potentiating peptides in viper venoms. For better insights into variations and potential impacts of medical significance, the venoms were compared against other Palearctic viper proteomes, including the first genus-wide Montivipera venom comparison. This will help the risk assessment of snakebite envenoming by these vipers and aid in predicting the venoms' pathophysiology and clinical treatments.

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“…are used). Alternatively, the observed differences in neutralising potencies could be due to the considerably higher abundance of α-NTxs in N. haje venom [54,57].…”

Section: Commercial Antivenomsmentioning

confidence: 99%

Anin vitroassay to investigate venom neurotoxin activity on muscle-type nicotinic acetylcholine receptor activation and for the discovery of toxin-inhibitory molecules

Patel

Clare

Ledsgaard

et al. 2023

Preprint

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Snakebite envenoming is a neglected tropical disease that causes over 100,000 deaths annually. Envenomings result in variable pathologies, but systemic neurotoxicity is among the most serious and is currently only treated with difficult to access and variably efficacious commercial antivenoms. Venom-induced neurotoxicity is often caused by α-neurotoxins antagonising the muscle-type nicotinic acetylcholine receptor (nAChR), a ligand-gated ion channel. Discovery of therapeutics targeting α-neurotoxins is hampered by relying on binding assays that do not reveal restoration of receptor activity or more costly and/or lower throughput electrophysiology-based approaches. Here, we report the validation of a screening assay for nAChR activation using immortalised TE671 cells expressing the γ-subunit containing muscle-type nAChR and a fluorescent dye that reports changes in cell membrane potential. Assay validation using traditional nAChR agonists and antagonists, which either activate or block ion fluxes, was consistent with previous studies. We then characterised antagonism of the nAChR by a variety of elapid snake venoms that cause muscle paralysis in snakebite victims, before defining the toxin-inhibiting activities of commercial antivenoms, and new types of snakebite therapeutic candidates, namely monoclonal antibodies, decoy receptors, and small molecules. Our findings show robust evidence of assay uniformity across 96-well plates and highlight the amenability of this approach for the future discovery of new snakebite therapeutics via screening campaigns. The described assay therefore represents a useful first-step approach for identifying α-neurotoxins and their inhibitors in the context of snakebite envenoming, and it should provide wider value for studying modulators of nAChR activity from other sources.

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Spatial Venomics─Cobra Venom System Reveals Spatial Differentiation of Snake Toxins by Mass Spectrometry Imaging

Cited by 10 publications

References 52 publications

Proteins from shrews’ venom glands play a role in gland functioning and venom production

Proteins from shrews’ venom glands play a role in gland functioning and venom production

Venomics and Peptidomics of Palearctic Vipers: A Clade-Wide Analysis of Seven Taxa of the Genera Vipera, Montivipera, Macrovipera, and Daboia across Türkiye

Anin vitroassay to investigate venom neurotoxin activity on muscle-type nicotinic acetylcholine receptor activation and for the discovery of toxin-inhibitory molecules

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