Venom Composition in a Phenotypically Variable Pit Viper (<i>Trimeresurus insularis</i>) across the Lesser Sunda Archipelago

Jones, Brenda Kathryn; Saviola, Anthony J.; Reilly, Sean B.; Stubbs, Alexander L.; Arida, Evy; Iskandar, Djoko T.; McGuire, Jimmy A.; Yates, John R.; Mackessy, Stephen P.

doi:10.1021/acs.jproteome.9b00077

Cited by 25 publications

(34 citation statements)

References 79 publications

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“…In general, two approaches are used in the analysis of venom proteomes. In the first one, the venom is freeze-dried or dried and suspended in a specific amount of buffer to obtain samples at a specific concentration [24][25][26][27]. The second approach uses crude venom and this approach requires a step to measure the protein concentration of the sample [28][29][30].…”

Section: Introductionmentioning

confidence: 99%

Comparison of Methods for Measuring Protein Concentration in Venom Samples

Bocian

Sławek

Jaromin

et al. 2020

Animals

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Simple Summary: Snake venom is mostly composed of proteins and peptides, which are of interest to many researchers due to their potential pharmacological properties. Due to their biochemical character, these components are analyzed using proteomic techniques such as electrophoresis, chromatography and mass spectrometry. A very important stage of such studies is the measurement of protein concentration in the sample, which is most often performed by colorimetric methods. In the presented article, we used five such techniques on venoms of two snake species, namely Agkistrodon contortrix and Naja ashei. In the case of A. contortrix venom, four methods provide similar concentration values, whereas, in the case of N. ashei, the differences between results are very significant. The source of these differences should probably be seen in the differences in amino acid composition of proteins of these two venoms. With this report, we would like to draw attention to the need to select an appropriate method for measuring the concentration of protein in the venom, especially in the case of Elapid species.Abstract: Snake venom is an extremely interesting natural mixture of proteins and peptides, characterized by both high diversity and high pharmacological potential. Much attention has been paid to the study of venom composition of different species and also detailed analysis of the properties of individual components. Since proteins and peptides are the active ingredients in venom, rapidly developing proteomic techniques are used to analyze them. During such analyses, one of the routine operations is to measure the protein concentration in the sample. The aim of this study was to compare five methods used to measure protein content in venoms of two snake species: the Viperids representative, Agkistrodon contortrix, and the Elapids representative, Naja ashei. The study showed that for A. contortrix venom, the concentration of venom protein measured by four methods is very similar and only the NanoDrop method clearly stands out from the rest. However, in the case of N. ashei venom, each technique yields significantly different results. We hope that this report will help to draw attention to the problem of measuring protein concentration, especially in such a complex mixture as animal venoms.

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Section: Introductionmentioning

confidence: 99%

Comparison of Methods for Measuring Protein Concentration in Venom Samples

Bocian

Sławek

Jaromin

et al. 2020

Animals

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“…Previous analysis of fibrinogen degradation on a very limited range of Trimeresurus species only provided qualitative, descriptive data and did not quantify the rates of fibrinogen chain cleavage (Jones et al, 2019). Further, this prior study did not attempt to determine the effect fibrinogen cleavage had upon clotting or the ability of antivenom to neutralise these pathological effects.…”

Section: Discussionmentioning

confidence: 99%

“…Further, this prior study did not attempt to determine the effect fibrinogen cleavage had upon clotting or the ability of antivenom to neutralise these pathological effects. In addition, prior studies also only measured antivenom binding through the use ELISA or Western Blot, which would reveal only simple antivenom binding to venom proteins, with no kinetic information, and, crucially prior studies did not ascertain whether the antivenom was effective in neutralising the venom's pathological functions (Jones et al, 2019;Tan et al, 2017a). In contrast, in this study we not only quantified the effects upon fibrinogen but also investigated the ability of the Thai Red Cross Green Pit Viper Antivenin to neutralise the coagulotoxic effects of the 13 species of Trimeresurus investigated here.…”

Section: Discussionmentioning

confidence: 99%

“…flavomaculatus, T. insularis, T. macrops, T. purpureomaculatus, and T. stejnegeri (Clark and Davidson, 1997;Jones et al, 2019;Mitrakul, 1973;Rojnuckarin et al, 1999;Tai et al, 2004;Tan et al, 2017a;Tan, 2010;Tan et al, 1994;Visudhiphan et al, 1989;Witharana et al, 2019;Wongtongkam et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

“…This variation has direct implications for the relative clinical usefulness of antivenoms against species not included in the immunising mixture, or varying populations of the species used (Debono et al, 2017a;Dobson et al, 2018;Fry et al, 2003a;Lister et al, 2017;Rogalski et al, 2017;Sousa et al, 2018;Sunagar et al, 2014;Tan et al, 2017a;Yang et al, 2016;Zdenek et al, 2019). Many current antivenomic strategies are mostly limited to the immunoreactivity of the antivenom towards the venom components, these do not ascertain whether the antivenom is effective in neutralising venom pathological functions (Jones et al, 2019;Williams et al, 2011;Xu et al, 2017). Out of the ten commonly described Gloydius species, antivenom exists only for four (G. blomhoffii, G. brevicaudus, G. halys and G. ussuriensis) ( Supplementary Table 6.1).…”

Section: Introductionmentioning

confidence: 99%

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A blood bond: evolutionary, pathophysiological and biodiscovery implications of coagulotoxic snake venoms

Debono¹

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Snakebite is a massive global burden and is a neglected area of tropical diseases. Snake venom neurotoxins have been the subject of intense research efforts, albeit significantly less is known about coagulotoxins. Venoms can deleteriously affect any physiological system reachable by the bloodstream, including being able to directly interfere with the coagulation cascade. Such coagulopathic toxins may be either anticoagulant or procoagulant and snake venoms are unique in their use of procoagulant toxins for predatory purposes. Our limited knowledge into the mechanisms of action of snake venom hinders our understanding of the clinical pathologies and also restricts our use of these powerful natural products as lead compounds in drug design and development. Two snake families which exhibit coagulant properties and lead to a range of clinical pathologies are vipers, Viperidae, and non-front-fanged snakes, Colubridae. The primary focus of this thesis was to investigate and compare the coagulotoxicity effects and mechanisms of various species of known venomous snakes that effect coagulation. Species chosen for this investigation have had limited previous research conducted, thus a knowledge gap in the literature remains. These coagulant properties were investigated through a multi-disciplinary approach, highlighting key results from both families.Data chapters 2 and 3 explore venom from the boomslang (Dispholidus typus) and the twig snakes (Thelotornis species) -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. We found that T. mossambicanus produced a significantly stronger coagulation response compared to D. typus, governed by strong prothrombin activation through PIII-SVMPs.Despite similarities in symptoms and clinical manifestations, venom composition differs widely between the two species, recovered from a combined approach of venomics and transcriptomics. 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. Envenomation by Crotalinae such as Asian pit vipers can induce multiple clinical complications resulting from coagulopathic and neuropathic effects. Data chapters 4 -8 investigate the Asian Pit Viper clade. While intense research has been undertaken for some species, such as Calloselasma rhodostoma, functional coagulopathic effects have been neglected. As these species' venoms affect the blood coagulation cascade and are known to produce haemorrhagic shock in envenomed patients, we investigated their effects upon coagulation using venoms of 33 species from the Asian pit viper clade including Azemiops, Calloselasma, Deinagkistrodon, Gloydius, Hypnale, Protobothrops, Trimeresurus and Tropidolaemus. Fibrinogen was the main coagulation target among th...

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Quantification of snake venom proteomes by mass spectrometry‐considerations and perspectives

Calvete

Lomonte

Saviola

et al. 2023

Mass Spectrometry Reviews

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The advent of soft ionization mass spectrometry‐based proteomics in the 1990s led to the development of a new dimension in biology that conceptually allows for the integral analysis of whole proteomes. This transition from a reductionist to a global‐integrative approach is conditioned to the capability of proteomic platforms to generate and analyze complete qualitative and quantitative proteomics data. Paradoxically, the underlying analytical technique, molecular mass spectrometry, is inherently nonquantitative. The turn of the century witnessed the development of analytical strategies to endow proteomics with the ability to quantify proteomes of model organisms in the sense of “an organism for which comprehensive molecular (genomic and/or transcriptomic) resources are available.” This essay presents an overview of the strategies and the lights and shadows of the most popular quantification methods highlighting the common misuse of label‐free approaches developed for model species' when applied to quantify the individual components of proteomes of nonmodel species (In this essay we use the term “non‐model” organisms for species lacking comprehensive molecular (genomic and/or transcriptomic) resources, a circumstance that, as we detail in this review‐essay, conditions the quantification of their proteomes.). We also point out the opportunity of combining elemental and molecular mass spectrometry systems into a hybrid instrumental configuration for the parallel identification and absolute quantification of venom proteomes. The successful application of this novel mass spectrometry configuration in snake venomics represents a proof‐of‐concept for a broader and more routine application of hybrid elemental/molecular mass spectrometry setups in other areas of the proteomics field, such as phosphoproteomics, metallomics, and in general in any biological process where a heteroatom (i.e., any atom other than C, H, O, N) forms integral part of its mechanism.

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Venom Composition in a Phenotypically Variable Pit Viper (Trimeresurus insularis) across the Lesser Sunda Archipelago

Cited by 25 publications

References 79 publications

Comparison of Methods for Measuring Protein Concentration in Venom Samples

Comparison of Methods for Measuring Protein Concentration in Venom Samples

A blood bond: evolutionary, pathophysiological and biodiscovery implications of coagulotoxic snake venoms

Quantification of snake venom proteomes by mass spectrometry‐considerations and perspectives

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