Comparative proteomic analysis of the venom of the taipan snake, Oxyuranus scutellatus, from Papua New Guinea and Australia: Role of neurotoxic and procoagulant effects in venom toxicity
Graphical abstract
Highlights
► The proteomes of venoms of taipan from Australia and Papua New Guinea were compared. ► Phospholipase A2 was the most abundant component in both venoms. ► Monospecific antivenom neutralized effectively venoms from the two populations. ► Neurotoxicity and thrombosis are relevant mechanisms for killing prey.
Introduction
Australia and New Guinea harbor a rich biodiversity of venomous snakes of the family Elapidae, which include species possessing some of the most toxic venoms in the world [1]. Among them, the taipans (Oxyuranus spp.), are distributed in Australia, Papua New Guinea and Indonesian Papua, and produce some of the world's most toxic snake venoms [2]. Combined with their agility, speed, large adult body size (up to 3 m for Oxyuranus scutellatus), high venom output (> 100 mg) and nervous dispositions, taipans are extremely dangerous snakes to encounter. Snakebites by Oxyuranus spp. often result in severe neurotoxic envenoming which may be fatal in the absence of prompt antivenom treatment [3], [4]. There are three species of taipans: (a) O. scutellatus, with populations in northern Australia, southern Papua New Guinea and southern Indonesian Papua [1], [5], [6]; (b) Oxyuranus microlepidotus, the inland taipan, which occurs in south-western Queensland and north-eastern to central-northern South Australia [7]; and (c) Oxyuranus temporalis, known from a handful of specimens collected in the south-eastern deserts of Western Australia [7]. O. scutellatus occurs in a wide variety of habitats including sugarcane fields and woodlands in the eastern and north coast of Australia and in savannah regions of southern New Guinea [5]. Phenotypic differences between the New Guinean and Australian populations of the taipan had prompted their classification as separate subspecies, Oxyuranus scutellatus canni and Oxyuranus s. scutellatus, respectively [8]. However, recent evidence from mitochondrial DNA analysis revealed high similarities between these populations, demonstrating very recent genetic exchange among them, thus questioning their subspecific status [6], [7]. Hence, further studies are required to assess the intraspecies variation between these populations.
Human envenomings by O. scutellatus are not common in northern Australia [1]. However, this species inflicts many severe bites in the southern regions of Papua New Guinea (PNG) [4]. These envenomings are predominantly characterized by a neurotoxic effect of rapid onset, which often leads to respiratory paralysis in the absence of timely administration of antivenom [3], [9]. In addition, coagulopathy associated with spontaneous bleeding has been described in these patients [10], together with myotoxicity and cardiac disturbances in some cases [11]. The toxins responsible for these effects have been isolated and characterized, such as the potent neurotoxic and myotoxic heterotrimeric phospholipase A2 (PLA2) complex taipoxin (named cannitoxin for the West Papuan population) [12], [13], two monomeric PLA2s, named OS1 and OS2 [14], Oscutarin-C, a prothrombin activator responsible for the characteristic coagulopathy [15], post-synaptically acting α-neurotoxins [16], and taicatoxin, a blocker of voltage-dependent calcium channels [17]. In addition, cDNA analysis derived from venom gland transcripts detected various putative toxin genes, including PLA2s, neurotoxins, cysteine-rich secretory proteins (CRISPs), a venom natriuretic peptide and a nerve growth factor [18]. However, besides the proteins previously characterized from this venom, the presence in the venom of these additional components encoded by these transcripts, and their possible biological activity, remain unknown.
The present work presents a comparative proteomic analysis of the venoms of O. scutellatus from Australia and PNG, in order to ascertain whether there are phenotypic differences in the venom composition of these populations. In parallel, the most relevant biological activities of these venoms were determined, and correlated with the venom proteome. Owing to the accelerated processes that characterize the evolution of snake venom toxins [19], [20], [21], it was of interest to assess whether the venom proteome of these very similar but allopatric populations differs, in order to predict possible variations in their biological/toxicological profiles, and in the immunologically cross-reactivity of antivenoms raised against these venoms. In addition, the potential role of neurotoxic and procoagulant components of these venoms in prey immobilization was explored in a mouse model.
Section snippets
Venom fractionation
The venom of O. scutellatus from PNG was a pool obtained from twelve healthy, adult specimens collected in PNG's Milne Bay Province and Central Province. These snakes were maintained in a purpose-built serpentarium at the University of PNG, and venom was collected at 21 day intervals. Venom was obtained using Parafilm-covered Eppendorf tubes. Samples contaminated by blood were discarded, and all samples were handled using plastic pipettes and tubes. Venom was snap-frozen to − 80 °C, before being
Venomics
The RP-HPLC elution profiles of O. scutellatus venoms from PNG and Australia, and the SDS-PAGE migration of the proteins present in each fraction are shown in Fig. 1, Fig. 2. Although the two chromatograms had a similar number of peaks, 23 and 25, the elution patterns revealed differences. Proteins related to seven and nine protein families were identified by mass spectrometry in the venoms of Australian and PNG O. scutellatus, respectively. These families of proteins identified in both venoms
Acknowledgments
The authors thank Dr Andrés Hernández and B.Sc. Julissa Fonseca (Instituto Clodomiro Picado) for their collaboration, as well as Dr Bruno Lomonte for performing some of the mass spectrometry analyses. This study was supported by CONARE, Vicerrectoría de Investigación (Universidad de Costa Rica) (projects 741-A7-611 and 741-A9-506), CRUSA-CSIC (Project 2009CR0021), Ministerio de Ciencia e Innovación, Madrid, Spain (grant BFU2010-17373), Generalitat Valenciana, Valencia, Spain (grant
References (66)
- et al.
The lethality in mice of dangerous Australian and other snake venoms
Toxicon
(1979) - et al.
Snakes across the Strait: Trans-Torresian phylogeographic relationships in three genera of Australasian snakes (Serpentes: Elapidae: Acantophis, Oxyuranus, and Pseudechis)
Mol Phylogenet Evol
(2005) - et al.
Electrocardiographic abnormalities in patients bitten by taipans (Oxyuranus scutellatus canni) and other elapid snakes in Papua New Guinea
Trans R Soc Trop Med Hyg
(1997) - et al.
Identification and properties of very high affinity brain membrane-binding sites for a neurotoxic phospholipase from the taipan venom
J Biol Chem
(1989) - et al.
Prothrombin activation by an activator from the venom of Oxyuranus scutellatus (Taipan snake)
J Biol Chem
(1986) - et al.
Variations in the pharmacological profile of post-synaptic neurotoxins isolated from the venoms of the Papuan (Oxyuranus scutellatus canni) and coastal (Oxyuranus scutellatus scutellatus) taipans
Neurotoxicology
(2010) - et al.
Isolation and physiological characterization of taicatoxin, a complex toxin with specific effects on calcium channels
Toxicon
(1992) - et al.
Molecular evolution of myotoxic phospholipases A2 from snake venom
Toxicon
(2003) - et al.
Evolution of an arsenal: structural and functional diversification of the venom system in the advanced snakes (Caenophidia)
Mol Cell Proteomics
(2008) - et al.
Pharmacological activities of a toxic phospholipase A isolated from the venom of the snake Bothrops asper
Comp Biochem Physiol C
(1986)
On the quaternary structure of taipoxin and textilotoxin: the advantage of being multiple
Toxicon
The effects of taipoxin and notexin on the function and fine structure of the murine neuromuscular junction
Neuroscience
The neurotoxicity of the venom phospholipases A2, notexin and taipoxin
Exp Neurol
Taipoxin, an extremely potent presynaptic snake venom neurotoxin. Elucidation of the primary structure of the acidic carbohydrate-containing taipoxin subunit, a phospholipase homolog
FEBS Lett
Receptors for a growing family of secreted phospholipases A2
Trends Pharmacol Sci
Feeding in snakes
Digestive properties of the venom of the Australian Coastal Taipan, Oxyuranus scutellatus (Peters, 1867)
Toxicon
Skeletal muscle degeneration induced by venom phospholipases A2: insights into the mechanisms of local and systemic myotoxicity
Toxicon
The intriguing world of prothrombin activators from snake venom
Toxicon
A novel small conductance Ca2 +-activated K+ channel blocker from Oxyuranus scutellatus taipan venom. Re-evaluation of taicatoxin as a selective Ca2 + channel probe
J Biol Chem
Proteome and immunome of the venom of the Thai cobra, Naja kaouthia
Toxicon
Structural considerations of the snake venom metalloproteinases, key members of the M12 reprolysin family of metalloproteinases
Toxicon
Characterization of mocarhagin, a cobra venom metalloproteinase from Naja mocambique mocambique, and related proteins from other Elapidae venoms
Toxicon
Purification, cloning and characterization of a metalloproteinase from Naja atra venom
Toxicon
Structures of two elapid snake venom metalloproteinases with distinct activities highlight the disulfide patterns in the D domain of ADAMalysin family proteins
J Struct Biol
Species specific sensitivity towards the hemorrhagin of Ophiophagus hannah (Elapidae)
Toxicon
A novel prothrombin activator from the venom of Micropechis ikaheka: isolation and characterization
Arch Biochem Biophys
Novel natriuretic peptides from the venom of the inland taipan (Oxyuranus microlepidotus): isolation, chemical and biological characterization
Biochem Biophys Res Commun
Characterization of ‘basparin A’, a prothrombin-activating metalloproteinase, from the venom of the snake Bothrops asper that inhibits platelet aggregation and induces defibrination and thrombosis
Arch Biochem Biophys
The mass of venom injected by two Elapidae: the taipan (Oxyuranus scutellatus) and the Australian tiger snake (Notechis scutatus)
Toxicon
Clinical toxicology of snakebite in Australia and New Guinea
Snake bites by the Papuan taipan (Oxyuranus scutellatus canni): paralysis, hemostatic and electrocardiographic abnormalities, and effects of antivenom
Am J Trop Med Hyg
Snakebite in Papua New Guinea
Cited by (56)
On characterizing the Red-headed Krait (Bungarus flaviceps) venom: Decomplexation proteomics, immunoreactivity and toxicity cross-neutralization by hetero-specific antivenoms
2022, Comparative Biochemistry and Physiology - Part D: Genomics and ProteomicsAnticoagulant Micrurus venoms: Targets and neutralization
2021, Toxicology LettersCoagulotoxic effects by brown snake (Pseudonaja) and taipan (Oxyuranus) venoms, and the efficacy of a new antivenom
2019, Toxicology in VitroCitation Excerpt :However, our examination of the well-neutralised Oxyuranus clade revealed no correlation between antivenom efficacy and clotting time (PGLS: df = 1, t = −0.0670, p = .9492). Herrera et al. (2012) found the coagulotoxicity of the PNG O. scutellatus to be greater than the Australian O. scutellatus population (and the latter was thus better neutralised by ICP antivenom in that study), but with a larger dataset there was no direct correlation between clotting time and antivenom efficacy. In addition to the larger Oxyuranus dataset in this study, Hererra et al. (2012) and others (Vargas et al., 2011) used a manual observation method (MCD: Minimal Coagulant Dose (Theakston and Reid, 1983)) to determine clotting time ‘by eye’ which resulted in large error bars, whereas this study used a robot-operated machine to measure clotting times, which resulted in remarkably small error bars (smaller than the line symbols in most cases).