Purification and partial characterization of an anticoagulant PLA₂ from the venom of Indian Daboia russelii that induces inflammation through upregulation of proinflammatory mediators

Abstract

The present study describes the purification and partial characterization of a basic anticoagulant PLA enzyme named as Rv(i) PLA from the venom of Indian Daboia russelii. The molecular mass of the protein was found to be 13,659.65 Da, and peptide mass fingerprinting revealed that it belongs to group II PLA family. The peptide sequence showed similarity to uncharacterized basic PLA enzyme having an accession no. of P86368 reported from Sri Lankan D. russelii. Rv(i) PLA exhibited strong phospholipase A and anticoagulant activity. It also induced expression of COX-2 and TNF-α mRNA in a dose-dependent manner in phorbol 12-myristate 13-acetate differentiated THP-1 cells, which play a crucial role during inflammation. Chemical modification of His residue in Rv(i) PLA with p-bromophenacyl bromide abolished the enzymatic, anticoagulant, and inflammatory activities. The result indicates that the catalytic site of Rv(i) PLA might play a vital role in inducing inflammation at the bite site during D. russelii envenomation.

Effect of Vipera ammodytes ammodytes Snake Venom on the Human Cytokine Network

“…These cytokines possess a pronounced proinflammatory effect and are capable of inducing the expression of several other cytokines and chemokines, including IL-8 [ 33 , 34 ]. Our findings partially correlate with data available in the literature, as increased levels of IL-1β have frequently been reported following administration of snake venoms [ 35 , 36 ], svPLA 2 s [ 13 , 37 ], or SVMPs [ 12 , 20 ]. However, the expression of IL-1α following envenomation with snake venoms has not yet been the focus of research.…”

“…The increase in TNF expression was to be expected, considering high levels of TNF-α have been observed in numerous experiments involving snake venoms. These experiments used either crude snake venoms [ 11 , 35 , 36 , 40 , 43 ] or isolated snake venom proteins, including SVMPs [ 19 , 20 , 57 ], svPLA 2 s [ 13 , 37 , 44 ], or LAAOs [ 23 , 58 ].…”

“…Arachidonic acid functions as substrate for the synthesis of various proinflammatory mediators, such as leukotrienes (LT), thromboxane A 2 (TXA 2 ), prostacyclin, and prostaglandins (PG). Thus, the inflammatory effect of svPLA 2 s can be directly linked to their enzymatic activity [ 11 , 12 , 13 ]. However, there are reports of catalytically inactive svPLA 2 s capable of inducing inflammation and nociceptive responses, suggesting the existence of other inflammation-inducing mechanisms not related to the arachidonic pathway [ 14 , 15 , 16 ].…”

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

“…The effect of erythrofibrase on coagulation of platelet-poor plasma (PPP) was measured by calculating recalcification time (RT) as described by Deka et al [ 41 ]. Different doses (0.01–10 µg/mL) of erythrofibrase were preincubated with PPP for 2 min at 37 °C.…”

“…The effect of erythrofibrase on coagulation of platelet-poor plasma (PPP) was measured by calculating recalcification time (RT) as described by Deka et al [41]. Different doses (0.01-10 µg/mL) of erythrofibrase were preincubated with PPP for 2 min at 37 • C. Clot formation was initiated by addition of 50 mM CaCl 2 , and change in OD was monitored at 405 nm every 10 s for 900 s using a MultiSkan GO Spectrophotometer (ThermoFisher Scientific, Waltham, MA, USA).…”