Proteomic insight into the venom composition of the largest European rear-fanged snake, Malpolon monspessulanus monspessulanus

Abstract

Snake envenomations constitute a worldwide neglected tropical disease, with the vast majority of lethal bites inflicted by front-fanged snakes from the viperid and elapid groups. Rear-fanged snakes (colubrids) were often considered harmless and as a result, are much less studied, but several documented deaths have suggested potent venom in this group as well. The largest European snake (Malpolon monspessulanus monspessulanus), known as the "Montpellier snake", is such a rear-fanged snake that belongs to the Lamprophiidae family. Its venom remains largely unknown but cases of envenomation with neurological symptoms have been reported. Here, we provide the first insights into the composition of its venom using mass spectrometry methods. First, liquid chromatography coupled mass spectrometry analysis of the manually collected venom samples reveals a complex profile, with the majority of masses encompassing the range 500-3000 Da, 4000-8000 Da, and 10 000-30 000 Da. Next, shotgun proteomics allowed the identification of a total of 42 different known families of proteins, including snake venom metalloproteinases, peptidase M1, and cysteine-rich secretory proteins, as the most prominent. Interestingly, three-finger toxins were not detected, suggesting that neurotoxicity may occur via other, yet to be determined, toxin types. Overall, our results provide the basis for a better understanding of the effects of a peculiar snake venom on human symptomatology, but also on the main prey consumed by this species.

Keywords: Montpellier's snake; Opistoglyph; Proteomics; Toxins.

Fig. 1

The Montpellier snake, Malpolon monspessulanus monspessulanus, from France in its natural environment (top panels show a male specimen, photo credit to Jean-Marie Ballouard). Bottom left panel shows the venom collection procedure (female specimen, photo credit to Sébastien Caron), facilitated by the presence of enlarged grooved teeth (bottom right panel).

Fig. 2

LC-MS analysis of Malpolon monspessulanus monspessulanus venom. The Total Ion Current (TIC) traces obtained from two representative venom samples are shown. Panel A was from the most complex sample, whereas panel B represents the most frequently observed venom profile. Inserts show the mass distribution for each sample.

Fig. 3

16 Snake venom protein families identified in the proteome of M. monspessulanus monspessulanus. SVMP: Snake Venom Metalloproteinase, PEPM1: Peptidase M1, PLA2: Phospholipase A2, CRISP: cysteine-rich secretory protein, NucPyro: Nucleotide pyrophosphatasephosphodiesterase, PL-B: Phospholipase B-like, GH30: Glycosyl hydrolase 30, GH56: Glycosyl hydrolase 56, NatPep: Natriuretic peptide, PGF: PDGFVEGF growth factor, PEPS1: Peptidase S1, VCCH: Venom complement C3 homolog.

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