Proteome Decomplexation of Trimeresurus erythrurus Venom from Mizoram, India

Abstract

Green pit vipers are the largest group of venomous vipers in tropical and subtropical Asia, which are responsible for most of the bite cases across this region. Among the green pit vipers of the Indian subcontinent, Trimeresurus erythrurus is the most prevalent; however, limited knowledge is available about its venomics. Proteome decomplexation of T. erythrurus venom using mass spectrometry revealed a blend of 53 different proteins/peptides belonging to 10 snake venom protein families. Phospholipase A 2 and snake venom serine proteases were found to be the major enzymatic families, and Snaclec was the major nonenzymatic family in this venom. These protein families might be responsible for consumptive coagulopathy in victims. Along with these, snake venom metalloproteases, L-amino acid oxidases, disintegrins, and cysteine-rich secretory proteins were also found, which might be responsible for inducing painful edema, tissue necrosis, blistering, and defibrination in patients. Protein belonging to C-type lectins, C-type natriuretic peptides, and glutaminyl-peptide cyclotransfreases were also observed as trace proteins. The crude venom shows platelet aggregation in the absence of any agonist, suggesting their role in alterations in platelet functions. This study is the first proteomic analysis of T. erythrurus venom, contributing an overview of different snake venom proteins/peptides responsible for various pathophysiological disorders obtained in patients. Data are available via ProteomeXchange with the identifier PXD038311.

Isolation and Functional Characterization of Erythrofibrase: An Alfa-Fibrinogenase Enzyme from Trimeresurus erythrurus Venom of North-East India

“…Crude venom of Trimeresurus erythrurus was firstly fractionated using reverse phase chromatography as previously described [ 17 ], using Symmetry C18 column (particle size 3.5 μm, a pore size 300 Å) with a linear gradient of 20−65% and 65–80% acetonitrile containing 0.1% ( v / v ) TFA for 20−100 min and 100−110 min, respectively. The elution was carried out at a flow rate of 0.8 mL/min at 215 nm, and fraction 7, showing highest amidolytic activity on substrates specific for thrombin (S2238, Chromgenix, Bedford, MA, USA) and plasmin (S2251, Chromgenix, Bedford, MA, USA), was collected.…”

“…Mass spectrometry of the purified protein was performed as previously described by Thakur et al [17], according to the method given by Kinter and Sherman [40]. The purified Rp-HPLC peak was firstly reduced by DTT in urea (prepared freshly in 50 mM Tris-HCl, pH 8.0) and subjected to in-solution trypsin digestion.…”

“…Further, painful edema, blistering, necrosis, haemorrhage and platelet alteration can be attributed to snake venom metalloproteases (SVMPs), L-amino acid oxidases and disintegrins [ 16 ]. Previous studies have reported the presence of these protein families in the venom of Indian spot-tailed pit viper Trimeresurus erythrurus [ 17 ]. Further, the clinical ineffectiveness of commercially available polyvalent antivenom in reversing the prolonged coagulopathy has been reported previously [ 14 ].…”