In vitro effects of Echis carinatus venom on the human plasma proteome

Abstract

Echis carinatus venom (EV) is a complex mixture of toxins that contribute to its lethality. EV proteolytic activity was analyzed by zymography, chromogenic assays, and SDS-PAGE. To understand the molecular mechanism of the envenomation, we investigated the in vitro effect of EV on human plasma proteins. We looked for EV protein substrates and their proteolytic fragments. We analyzed EV proteolytic activity on standard proteins such as prothrombin or fibrinogen. To set up the optimal EV:plasma protein ratio conditions, plasma was incubated with EV (treated plasma), depleted of abundant proteins, and subjected to SDS-PAGE. Samples from control and treated plasma were also analyzed by 2-DE/MALDI-TOF MS, leading to the identification of four classes of plasma proteins cleaved by EV: proteases, protease inhibitors, binding proteins, and transporters. EV mainly proteolyzes entire proteins but can also act on physiological fragments. In summary, the physiological effects of EV proteases involve other important processes in addition to blood coagulation; complement activation and hemoglobin metabolism are also affected. In particular, the cleavage of protease inhibitors appears to be the mechanism through which the venom neutralizes the body's defenses.

Antihyaluronidase and Alkaline Phosphatase (ALP) Activities of Medicinal Plants to Combat Echis carinatus Venom-Induced Toxicities

“…Echis carinatus snake has around 0.6 m length, flat body, pointed tail, and is known as a true viper [7,12]. Echis carinatus envenoming effects anticoagulant or procoagulant activity due to presence of active enzymes in its venom which in turn disturbs the hemostatic system [13,14]. Envenomation of Echis also cause local tissue damage and cell necrosis by the synergistic effect of hydrolytic enzymes hyaluronidases, phospholipases A 2 , and proteases [10].…”

The Role of Conventional Two-Dimensional Electrophoresis (2DE) and Its Newer Applications in the Study of Snake Venoms

“…To investigate the effects of Echis carinatus envenomation on the human plasma proteome, an in vitro model utilizing 2DE as one of its core techniques has been established in two studies Guerranti et al, 2010). The results from these two studies showed that 2DE was capable of demonstrating global proteomic changes when the human plasma was incubated with the E. carinatus venom.…”

In vivo evaluation of homeostatic effects of Echis carinatus snake venom in Iran

“…In the past, it was wrongly assumed that a single ophidian species would generally cause either local or systemic effects and that vipers caused local and/or hemorrhagic effects, while elapids caused purely systemic, non-hemorrhagic effects [6-9]. …”