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. 2020 Jun 23:7:100048.
doi: 10.1016/j.toxcx.2020.100048. eCollection 2020 Sep.

Comparative proteomics of geographically distinct saw-scaled viper (Echis carinatus) venoms from India

Siddharth Bhatia  1 , Karthikeyan Vasudevan  1
Affiliations

Comparative proteomics of geographically distinct saw-scaled viper (Echis carinatus) venoms from India

Siddharth Bhatia et al. Toxicon X. .

Abstract

Snakebite is a socio-economic problem in tropical countries and it is exacerbated by geographical venom variation of snakes. We investigated on venom variation in geographically distinct populations of Echis carinatus from three ecologically distinct regions: Tamil Nadu (ECVTN), Goa (ECVGO), and Rajasthan (ECVRAJ). Venom was fractionated by RP-HPLC, combined with SDS-PAGE, and subjected to tandem mass spectrometry. Toxins were identified, and their relative abundance was estimated. Using NCBI database of Echis genus, we queried the MS/MS spectra, and found 69, 38 and 38 proteins in ECVTN, ECVGO and ECVRAJ respectively, belonging to 8-10 different toxin families. The differences in the venom profiles were due to change in the relative composition of the toxin families. Snake venom metalloproteinase (svMP), Snaclecs and Phospholipase A2 (PLA2) were the major venom components in all the venoms. Heteromeric Disintegrins were found in ECVTN and absent in other venoms. ECVRAJ showed higher abundance of low-molecular-weight (>30 kDa) proteins than ECVTN and ECVGO. Cysteine-rich venom protein (CRISP) was highest in ECVRAJ (7.34%), followed by ECVTN (0.01%) and in ECVGO, it was not detected. These findings highlight the need for evaluating the efficacy of the polyvalent anti-venom to neutralize the toxins from geographically distinct venoms of E. carinatus.

Keywords: De-complexation; Envenomation; Mass spectrometry; RP-HPLC; Snake venom; Toxins; Venomics.

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Conflict of interest statement

The authors have no conflict of interest. The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Figures

Image 1
Graphical abstract
Fig. 1
Fig. 1
20μg of Crude ECV from Tamil Nadu (TN), Goa (GO) and Rajasthan (RA) was loaded on 12% SDS-PAGE under reducing and non-reducing condition. Bands from each lane were cut, in-gel digested and subjected to MS/MS to identify toxin families and compared across lanes.
Fig. 2
Fig. 2
Reverse phase-HPLC of ECV from Tamil Nadu (ECVTN). 2–3 mg of venom was loaded and 30 fractions were collected manually, dried and reconstituted with double distilled water.
Fig. 3
Fig. 3
Reverse phase-HPLC of Echis carinatus venom from Goa (ECVGO). 2–3 mg of venom was loaded and 26 fractions were collected manually, dried and reconstituted with double distilled water.
Fig. 4
Fig. 4
Reverse phase-HPLC of Echis carinatus venom from Udaipur, Rajasthan (ECVRAJ). 2–3 mg of venom was loaded and 32 fractions were collected manually, dried and reconstituted with double distilled water.
Fig. 5
Fig. 5
Pie chart representing relative abundance of toxin families for Echis carinatus venom. svMP (snake venom Metalloproteinases), NGF (Neural growth factors), Phospholipase A2 (PLA2), Phospholipase B(PLB), renin-like aspartic protease (AsP), svSP (snake venom Serine proteinase), DIS domain (P-II svMP)- Disintegrin domain of P-II svMP, DC domain (Disintegrin-like and cysteine domain of P-III svMP), HYAL (Hyaluronidase), LAAO (L-amino acid oxidase), PLB (Phospholipase B), CRISP (Cysteine-rich secretory protein).
See this image and copyright information in PMC

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