Canopy Venom: Proteomic Comparison among New World Arboreal Pit-Viper Venoms

Debono, Jordan; Cochran, Chip; Kuruppu, Sanjaya; Nouwens, Amanda; Rajapakse, Niwanthi W.; Kawasaki, Minami; Wood, K; Dobson, James; Baumann, Kate; Jouiaei, Mahdokht; Jackson, Timothy N. W.; Koludarov, Ivan; Low, Dolyce H. W.; Ali, Syed Abid; Smith, Allan E.; Barnes, Andrew

doi:10.3390/toxins8070210

Cited by 8 publications

(4 citation statements)

References 126 publications

(202 reference statements)

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“…In addition to differential complexity, the venoms were functionally variable relative to each other in non-procoagulant activities, with D. typus having serine protease activity ( Figure 5 ), while conversely T. mossambicanus had strong PLA 2 activity ( Figure 6 ). It is known that evolution among arboreal specialists further influences venom composition of toxins’ families [ 48 ]. This is usually in the form of prey ecology, dependent on niche occupation.…”

Section: Discussionmentioning

confidence: 99%

“…Our proteomic investigations including using 1D and 2D SDS-PAGE, protein band purification and crude venom Shotgun analysis. They were performed as previously described by us [ 48 ] with the exception of additional LC-MS/MS analysis, following protein extraction from isolated 1D gel bands (described below in Section 4.4.1 and Section 4.4.2 ). Shotgun samples were analyzed using the methods described below ( Section 4.4.3 ).…”

Section: Methodsmentioning

confidence: 99%

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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

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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.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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

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“…The molecular biology, evolution, and pathology of venoms have long been fertile areas for research, particularly animal venoms of clinical significance. Some of this work has attempted to interpret biochemical and molecular evolutionary findings in the context of ecology, for instance, differences in venom composition being explained by some attributes that differ among species such as diet e.g., [ 1 ]. However, much rarer is an explicit attempt to study the evolutionary ecology of venom beyond simply using the ecological aspects as an ad hoc and post hoc explanatory framework.…”

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confidence: 99%

Special Issue: Evolutionary Ecology of Venom

Arbuckle

2021

Toxins

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“…Our proteomic investigations included using a combined approach of 1D and 2D SDS-PAGE, excision of gel bands and spots, and LC-MS/MS identification of the proteins therein. Proteomic methods were performed as previously described by us [21,140,141]. Figure 2.5A shows bands picked for each species for MSMS processing.…”

Section: Proteomicsmentioning

confidence: 99%

Novelty in three-finger toxin evolution

Dashevsky¹

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Snakes are a striking taxon on many levels and have an indelible place in the collective culture of humanity. This is due to a number of unusual features including their limblessness, ability to ingest large prey, but most importantly their venom. From a biological perspective, snakes are equally fascinating, showing how a simple body plan can be used to great effect in an extremely broad variety of niches. Snakes are globally distributed and ecologically diverse; as a result, since recent research has shown that most snakes are venomous to some degree, their venom is equally diverse. This thesis focuses on one major toxin family: three-finger toxins (3FTx). These toxins can be found in virtually all venomous snake lineages and are often neurotoxic. Since their original recruitment there have been numerous instances of structural and functional innovation within this toxin family. The research in this thesis pertains to several such novel adaptations in snakes of the colubrid and elapid families, many of whose venom is composed primarily of these toxins. Chapter 2 deals with the ancestral version 3FTx and how, in the colubrine genus Boiga, simple mutations in toxins can lead to structural changes, in this case additional cysteines enable the formation of dimeric toxins with increased potency. These novel toxins can, in turn, open up new niches to the snakes which possess those toxins. In B. irregularis they facilitated a destructive invasion event and may have similarly assisted in the spread of an ancestral Boiga since our research shows that several other species possess similar dimeric toxins. This echoes a process that occurred in the ancestor of the elapids where the loss of a pair of cysteines and their disulfide bond was followed by an incredible diversification of these toxins and the snakes that produce them. Chapter 3 explores venom composition and activity of the genus Calliophis, the most basal genus of the elapids. C. bivirgatus and C. intestinalis both possess extremely elongated venom glands and each produce 3FTx with novel sodium channel toxicity. However, the two venoms produce opposite effects from one another. These results suggests some intriguing possibilities about the evolutionary history of these novel traits in Calliophis and the general composition of their venoms inform our understanding of how the venoms of more derived elapids evolved.Chapters 4 and 5 focus on the genus Micrurus. This genus from the Americas is by far the most speciose genus of elapids and belongs to the coral snake clade which is the next branch in the phylogeny of the elapids after Calliophis. The venoms of these snakes all contain a high number of unique 3FTx, though some venoms may be dominated by PLA2 toxins in terms of overall composition. Chapter 4 focuses on the sequence diversity of 3FTx from this genus and the selection regimes under which they operate. This diversity may lay the groundwork for rapid changes in the toxin composition of the venom and accompanying changes in venom activity. Chapter 5 directly ...

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Canopy Venom: Proteomic Comparison among New World Arboreal Pit-Viper Venoms

Cited by 8 publications

References 126 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)

Special Issue: Evolutionary Ecology of Venom

Novelty in three-finger toxin evolution

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