Comparative Analysis of the Venom Proteomes of Vipera ammodytes ammodytes and Vipera ammodytes meridionalis

Abstract

The venom proteomics of Vipera ammodytes ammodytes and Vipera ammodytes meridionalis, snakes of public health significance and the most poisonous reptiles in Europe, were analyzed by FPLC, 2-D electrophoresis, sequence analysis, and MS/MS. FPLC analysis showed the presence of l-amino acid oxidase, monomeric and heterodimeric phospholipases A2, C-type lectin protein, and proteinases in the venom of V. a. ammodytes. Representatives of the same protein families were found in the venom of the other subspecies, V. a. meridionalis. N-terminally identical PLA2 neurotoxins were identified in both venoms. Difference in the PLA2 compositions of the venoms was also observed: a monomeric protein with phospholipase A2 activity, identical in the first 20 amino acid residues to the catalitically inactive acidic component of the heterodimeric PLA2 present in both venoms, was found only in that of V. a. meridionalis. Probably, this protein represents an intermediate form of the two components of the heterodimer. 2-D electrophoresis and MS/MS analysis showed that the two venoms shared a number of protein families: monomeric and heterodimeric Group II PLA2s, serine proteinases, Group I, II, and III metalloproteinases, l-amino acid oxidases (LAAOs), cysteine-rich secretory proteins, disintegrins, and growth factors. Totally, 38 venom components of the V. a. ammodytes, belonging to 9 protein families, and 67 components of the V. a. meridionalis venom belonging to 8 protein families were identified. The venom proteome of V. a. ammodytes shows larger diversity of proteins (139) in comparison to that of V. a. meridionalis (104 proteins). Most of the proteins are homologues of known representatives of the respective protein families. The protein compositions explain clinical effects of the V. ammodytes snakebites, such as difficulties in the breathing, paralysis, apoptosis, cloting disorders, hemorrhage, and tissue necrosis. The lists of secreted proteins by the two vipers can be used for further study of structure-function relationships in the toxins and for prediction and treatment of snakebite consequences.

Venomics of Vipera berus berus to explain differences in pathology elicited by Vipera ammodytes ammodytes envenomation: Therapeutic implications

“…Most SVSPs are N-or O-glycosylated, but to different extents. As reported for Vaa venom [19], diverse SVSPs are abundant in Vbb venom also (Table 1). Vbb SVSPs, with apparent molecular masses of 25, 35, 40 and 55 kDa, were identified in PRLP-S fractions 8 to 14 by similarity to SVPSs from related snakes, Vaa, Macrovipera lebetina (M. lebetina), Crotalus atrox (C. atrox), Agkistrodon contortrix contortrix (A. c. contortrix) and Trimeresurus stejnegeri (T. stejnegeri).…”

“…Nevertheless, in particular areas, for example in Mediterranean countries, snake venom poisoning represents considerable public health issue arguing for further improvements [4,5]. Venom proteomes of two European viper subspecies, Vaa and Vipera ammodytes meridionalis (Vam) have been reported so far [19]. The venom proteome of Vbb is presented in this work.…”

Comparative structural studies of two natural isoforms of ammodytoxin, phospholipases A2 from Vipera ammodytes ammodytes which differ in neurotoxicity and anticoagulant activity☆

“…The venom proteome of long-nosed viper Vipera ammodytes ammodytes was recently analyzed and proteins belonging to several different toxin families were identified, including a large diversity of phospholipases A 2 , L-amino acid oxidases, C-type lectins, serineproteinases, metalloproteinases, cysteine-rich secretory proteins, desintegrins, and growth factors (Georgieva et al, 2008). Isoforms A, B, and C of basic ammodytoxins (Atx) from the venom of V. a. ammodytes are potent presynaptically neurotoxic GIIA sPLA 2 s (Gubensek et al, 1980;Ritonja and Gubensek, 1985) capable of blocking the release of acetylcholine from peripheral neurons at the neuromuscular junction (Lee et al, 1984).…”

Snake Venomics, Antivenomics, and Venom Phenotyping: The Ménage à Trois of Proteomic Tools Aimed at Understanding the Biodiversity of Venoms

“…4.3) (Alape-Girón et al, 2008;Angulo et al, 2008;Bazaa et al, 2005;Calvete et al, 2007bCalvete et al, , c, 2009bGutiérrez et al, 2008;Juárez et al, 2004Juárez et al, , 2006Lomonte et al, 2008;Núñez et al, 2009;Sanz et al, 2006Sanz et al, , 2008aTashima et al, 2008;Wagstaff et al, 2009). A few other laboratories have also reported qualitative proteomic studies on several venoms, including those from the European vipers, Vipera ammodytes ammodytes and Vipera ammodytes meridionalis (Georgieva et al, 2008), Vipera aspis (Ferquel et al, 2007), Asian Daboia russelli siamensis (Risch et al, 2009), Amazonian Bothrops atrox (Guércio et al, 2006), and Brazilian Bothrops jararaca (Fox et al, 2006) and Bothrops insularis (Valente et al, 2009). These studies have allowed the inference of phylogenetic alliances within genera Bitis (Calvete et al, 2007c) and Sistrurus ; to rationalize the envenomation profiles of Atropoides and Bothrops species; to reveal gene regulation effects on venom protein expression in Sistrurus rattlesnakes (Gibbs et al, 2009); to correlate geographic intraspecific venom composition variation and reappraisal of Vipera aspis venom neurotoxicity (Ferquel et al, 2007); and to define venom-associated taxonomic markers (Tashima et al, 2008).…”