What killed Karl Patterson Schmidt? Combined venom gland transcriptomic, venomic and antivenomic analysis of the South African green tree snake (the boomslang), Dispholidus typus

Plá, Davinia; Sanz, Libia; Whiteley, Gareth; Wagstaff, Simon C.; Harrison, Robert A.; Casewell, Nicholas R.; Calvete, Juan J.

doi:10.1016/j.bbagen.2017.01.020

Cited by 58 publications

(80 citation statements)

References 84 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The presence of PLA 2 Type IIE is a significant discovery as it is the third type of PLA 2 to be characterised in snake venoms [44]. The molecular weights of identified P-III SVMP correspond to previous transcriptome and proteome data for D. typus [24]. …”

Section: Resultsmentioning

confidence: 81%

Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus)

Debono

Dobson

Casewell

et al. 2017

Toxins

View full text Add to dashboard Cite

Venoms can deleteriously affect any physiological system reachable by the bloodstream, including directly interfering with the coagulation cascade. Such coagulopathic toxins may be anticoagulants or procoagulants. Snake venoms are unique in their use of procoagulant toxins for predatory purposes. The boomslang (Dispholidus typus) and the twig snakes (Thelotornis species) are iconic African snakes belonging to the family Colubridae. Both species produce strikingly similar lethal procoagulant pathologies. Despite these similarities, antivenom is only produced for treating bites by D. typus, and the mechanisms of action of both venoms have been understudied. In this study, we investigated the venom of D. typus and T. mossambicanus utilising a range of proteomic and bioactivity approaches, including determining the procoagulant properties of both venoms in relation to the human coagulation pathways. In doing so, we developed a novel procoagulant assay, utilising a Stago STA-R Max analyser, to accurately detect real time clotting in plasma at varying concentrations of venom. This approach was used to assess the clotting capabilities of the two venoms both with and without calcium and phospholipid co-factors. We found that T. mossambicanus produced a significantly stronger coagulation response compared to D. typus. Functional enzyme assays showed that T. mossambicanus also exhibited a higher metalloprotease and phospholipase activity but had a much lower serine protease activity relative to D. typus venom. The neutralising capability of the available boomslang antivenom was also investigated on both species, with it being 11.3 times more effective upon D. typus venom than T. mossambicanus. In addition to being a faster clotting venom, T. mossambicanus was revealed to be a much more complex venom composition than D. typus. This is consistent with patterns seen for other snakes with venom complexity linked to dietary complexity. Consistent with the external morphological differences in head shape between the two species, CT and MRI analyses revealed significant internal structural differences in skull architecture and venom gland anatomy. This study increases our understanding of not only the biodiscovery potential of these medically important species but also increases our knowledge of the pathological relationship between venom and the human coagulation cascade.

show abstract

Section: Resultsmentioning

confidence: 81%

Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus)

Debono

Dobson

Casewell

et al. 2017

Toxins

View full text Add to dashboard Cite

show abstract

“…PLA 2 toxins, 13-14 kDa in size). However, given that D. typus is the only colubrid venom investigated in this study, and that colubrid PLA 2 s are distinct from those of vipers (24), this hypothesis requires testing in future work. Nonetheless, these findings are interesting because there are no prior reports of D. typus venom exhibiting anticoagulant activity, likely because this activity is masked by the net procoagulant activity of this venom (7).…”

Section: Resultsmentioning

confidence: 99%

“…The procoagulant peak could be correlated to a mass of ∼23 kDa (Table 2). This toxin most likely belongs to the SVMPs family, which are the most dominant toxin type in D. typus venom (24), and have previously been described to be responsible for the potent procoagulant activity of this species (63).…”

Section: Resultsmentioning

confidence: 99%

“…Using the data obtained for the tryptic digests of the samples, protein identification was carried out using MASCOT (Matrix Science, London, United Kingdom) searches against Swiss-Prot, NCBInr and species-specific databases. The latter were generated from previously published transcriptomic data for those species that were available (24–27) and were used for the identification of venom toxins for which activity was observed. The following search parameters were used: ESI-QUAD-TOF as the instrument type, semiTrypsin as the digestion enzyme allowing for one missed cleavage, carbamidomethyl on cysteine as a fixed modification, amidation (protein C-terminus) and oxidation on methionine as variable modifications, ± 0.05 Da fragment mass tolerance and ± 0.2 Da peptide mass tolerance.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

High throughput screening and identification of coagulopathic snake venom proteins and peptides using nanofractionation and proteomics approaches

Slagboom

Mladic

Xie³

et al. 2019

Preprint

Self Cite

View full text Add to dashboard Cite

17Snakebite is a neglected tropical disease that results in a variety of systemic and local pathologies in 18 envenomed victims and is responsible for around 138,000 deaths every year. Many snake venoms cause 19 severe coagulopathy that makes victims vulnerable to suffering life-threating haemorrhage. The 20 mechanisms of action of coagulopathic snake venom toxins are diverse and can result in both anticoagulant 21 and procoagulant effects. However, because snake venoms consist of a mixture of numerous protein and 22 peptide components, high throughput characterizations of specific target bioactives is challenging. In this 23 study, we applied a combination of analytical and pharmacological methods to identify snake venom toxins 24 from a wide diversity of snake species that perturb coagulation. To do so, we used a high-throughput 25 screening approach consisting of a miniaturised plasma coagulation assay in combination with a venom 26 nanofractionation approach. Twenty snake venoms were first separated using reversed-phase liquid 27 chromatography, and a post-column split allowed a small fraction to be analyzed with mass spectrometry, 28 while the larger fraction was collected and dispensed onto 384-well plates before direct analysis using a 29 plasma coagulation assay. Our results demonstrate that many snake venoms simultaneously contain both 30 procoagulant and anticoagulant bioactives that contribute to coagulopathy. In-depth identification analysis 31 from seven medically-important venoms, via mass spectrometry and nanoLC-MS/MS, revealed that 32 phospholipase A 2 toxins are frequently identified in anticoagulant venom fractions, while serine protease 33 and metalloproteinase toxins are often associated with procoagulant bioactivities. The nanofractionation 34 and proteomics approach applied herein seems likely to be a valuable tool for the rational development of 35 next-generation snakebite treatments by facilitating the rapid identification and fractionation of 36 coagulopathic toxins, thereby enabling specific targeting of these toxins by new therapeutics such as 37 monoclonal antibodies and small molecule inhibitors. Classified Personnel Information 39Author summary 40 Snakebite is a neglected tropical disease that results in more than 100,000 deaths every year. 41Haemotoxicity is one of the most common signs of systemic envenoming observed after snakebite, and 42 many snake venoms cause severe impairment of the blood coagulation that makes victims vulnerable to 43 suffering life-threating hemorrhage. In this study, we applied a combination of analytical and 44 pharmacological methods to identify snake venom toxins from a wide diversity of snake species that 45 interfere with blood coagulation. Twenty snake venoms were screened for their effects on the blood 46 coagulation cascade and based on the initial results and the medical relevance of the species, seven 47 venoms were selected for in-depth analysis of the responsible toxins using advanced identification 48 techniques. Our findings reveal a number ...

show abstract

“…However, while all three classes of SVMPs have been described from viper venoms, only P-III SVMPs have been detected in elapid and "colubrid" venoms . While these P-III SVMPs are typically relatively lowly abundant venom components in elapid snakes (e.g., <10% of venom toxins), they can be major components in "colubrids" (Mackessy and Saviola, 2016;Pla et al, 2017;Tasoulis and Isbister, 2017;Modahl et al, 2018a). These abundance differences likely underpin the distinct pathologies observed following envenomings by snakes found in these families.…”

Section: Snake Venom Metalloproteinases (Svmps)mentioning

confidence: 99%

Multifunctional Toxins in Snake Venoms and Therapeutic Implications: From Pain to Hemorrhage and Necrosis

et al. 2019

Self Cite

View full text Add to dashboard Cite

Animal venoms have evolved over millions of years for prey capture and defense from predators and rivals. Snake venoms, in particular, have evolved a wide diversity of peptides and proteins that induce harmful inflammatory and neurotoxic effects including severe pain and paralysis, hemotoxic effects, such as hemorrhage and coagulopathy, and cytotoxic/myotoxic effects, such as inflammation and necrosis. If untreated, many envenomings result in death or severe morbidity in humans and, despite advances in management, snakebite remains a major public health problem, particularly in developing countries. Consequently, the World Health Organization recently recognized snakebite as a neglected tropical disease that affects ∼2.7 million p.a. The major protein classes found in snake venoms are phospholipases, metalloproteases, serine proteases, and three-finger peptides. The mechanisms of action and pharmacological properties of many snake venom toxins have been elucidated, revealing a complex multifunctional cocktail that can act synergistically to rapidly immobilize prey and deter predators. However, despite these advances many snake toxins remain to be structurally and pharmacologically characterized. In this review, the multifunctional features of the peptides and proteins found in snake venoms, as well as their evolutionary histories, are discussed with the view to identifying novel modes of action and improving snakebite treatments.

show abstract

What killed Karl Patterson Schmidt? Combined venom gland transcriptomic, venomic and antivenomic analysis of the South African green tree snake (the boomslang), Dispholidus typus

Cited by 58 publications

References 84 publications

Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus)

Coagulating Colubrids: Evolutionary, Pathophysiological and Biodiscovery Implications of Venom Variations between Boomslang (Dispholidus typus) and Twig Snake (Thelotornis mossambicanus)

High throughput screening and identification of coagulopathic snake venom proteins and peptides using nanofractionation and proteomics approaches

Multifunctional Toxins in Snake Venoms and Therapeutic Implications: From Pain to Hemorrhage and Necrosis

Contact Info

Product

Resources

About