Purification and characterization of a platelet aggregation inhibitor acidic phospholipase A2 from Indian saw-scaled viper (Echis carinatus) venom

Kemparaju, K.; Krishnakanth, T.P.; Gowda, T. Veerabasappa

doi:10.1016/s0041-0101(99)00104-x

Cited by 34 publications

(11 citation statements)

References 29 publications

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“…The sequences of two peptides obtained by trypsin digestion matched the sequence of Russell's viper PLA 2 (Daboia russellii), while the venom we used was certified to be from E. carinatus sochureki. This may be explained by the fact that only one sequence of E. carinatus PLA 2 is deposited in the data base, whereas various isoenzymes are known in literature (13)(14)(15)27), and EV varies with country of origin and other factors (28) as shown in our experiments and declared by our supplier. Since the genera Echis and Daboia are very similar and both belong to the subfamily Viperinae, the E. carinatus PLA 2 that we analyzed may be a genetic variant, the sequence of which is closest to that of Russell's viper.…”

Section: Discussionmentioning

confidence: 84%

“…EV contains proteins with different toxic properties including opposite effects on blood clotting. Well known proteins are: disintegrins EC3 (5), EC6 (6), and echistatin (7), which inhibit the interaction of fibrinogen with the glycoprotein IIb-IIIa receptor on the platelet surface; echicetin (8) and ECLVIX/Xbp (9) with an opposite effect on platelet aggregation; two metalloproteases, ecarin (10,11) and carinactivase (12), which are prothrombin activators and act as procoagulant enzymes; and phospholipase A 2 (PLA 2 ) (13)(14)(15), the most abundant enzyme, which has many effects including inhibition of prothrombin activation by ecarin and carinactivase (16). When injected into mice, this complex mixture of proteins induces disseminated intravascular coagulation leading to death in less than a day.…”

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confidence: 99%

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Proteins from Mucuna pruriens and Enzymes fromEchis carinatus Venom

Guerranti¹,

Aguiyi²,

Neri³

et al. 2002

Journal of Biological Chemistry

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Mucuna pruriens seeds have been widely used against snakebite in traditional medicine. The antivenin property of a water extract of seeds was assessed in vivo in mice. The serum of mice treated with extract was tested for its immunological properties. Two proteins of Echis carinatus venom with apparent molecular masses of 25 and 16 kDa were detected by Western blot analysis carried out using IgG of mice immunized with extract or its partially purified protein fractions. By enzymatic in-gel digestion and electrospray ionization-mass spectrometry/mass spectrometry analysis of immunoreactive venom proteins, phospholipase A 2, the most toxic enzyme of snake venom, was identified. These results demonstrate that the observed antivenin activity has an immune mechanism. Antibodies of mice treated with non-lethal doses of venom reacted against some proteins of M. pruriens extract. Proteins of E. carinatus venom and M. pruriens extract have at least one epitope in common as confirmed by immunodiffusion assay.Snakebite is a considerable problem in certain tropical and subtropical countries. According to World Health Organization estimates, 40,000 of 5 million cases of snakebite are fatal. Antivenins obtained from horses treated with snake venom are one of the principal remedies against snakebite. This therapy has the disadvantage that antivenins must be given immediately, and snakebite victims may develop an adverse reaction including anaphylactic shock (1). The use of endogenous plants with a reputation against snakebite is therefore worth considering (2).In preliminary experiments (3, 4) we demonstrated that extract of M. pruriens (MPE), 1 a medicinal plant widely used in Nigeria for its chemical and pharmacological properties, protects mice against the lethal effect of Echis carinatus venom (EV). Both MPE and EV are heterogeneous mixtures, their interaction represents a complex phenomenon, and there is no information about its biochemical mechanism. EV contains proteins with different toxic properties including opposite effects on blood clotting. Well known proteins are: disintegrins EC3 (5), EC6 (6), and echistatin (7), which inhibit the interaction of fibrinogen with the glycoprotein IIb-IIIa receptor on the platelet surface; echicetin (8) and ECLVIX/Xbp (9) with an opposite effect on platelet aggregation; two metalloproteases, ecarin (10, 11) and carinactivase (12), which are prothrombin activators and act as procoagulant enzymes; and phospholipase A 2 (PLA 2 ) (13-15), the most abundant enzyme, which has many effects including inhibition of prothrombin activation by ecarin and carinactivase (16). When injected into mice, this complex mixture of proteins induces disseminated intravascular coagulation leading to death in less than a day. The composition of the M. pruriens seed is also complex and variable, with 20 -30% protein (lectins, globulins, protease inhibitors), 1-10% fat, 4 -5% ash, 4 -9% water, 4 -7% fiber (17-20), and L-DOPA (21), an interesting non-protein component. The aim of the present study was to stud...

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Section: Discussionmentioning

confidence: 84%

mentioning

confidence: 99%

Proteins from Mucuna pruriens and Enzymes fromEchis carinatus Venom

Guerranti¹,

Aguiyi²,

Neri³

et al. 2002

Journal of Biological Chemistry

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“…Numerous snake venom sPLA 2 s that modulate platelet function have been characterized [13,14,15,16,17,18,19] and different mechanisms of action shown [6,15,20,21,22,23,24,25,26]. The sPLA 2 s effect on platelet aggregation can be independent or dependent on their catalytic activity.…”

Section: Introductionmentioning

confidence: 99%

Interactions of PLA2-s from Vipera lebetina, Vipera berus berus and Naja naja oxiana Venom with Platelets, Bacterial and Cancer Cells

Samel

Vija

Kurvet

et al. 2013

Toxins

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Secretory phospholipasesA2 (sPLA2s) form a large family of structurally related enzymes widespread in nature. Herein, we studied the inhibitory effects of sPLA2s from Vipera lebetina (VLPLA2), Vipera berus berus (VBBPLA2), and Naja naja oxiana (NNOPLA2) venoms on (i) human platelets, (ii) four different bacterial strains (gram-negative Escherichia coli and Vibrio fischeri; gram-positive Staphylococcus aureus and Bacillus subtilis) and (iii) five types of cancer cells (PC-3, LNCaP, MCF-7, K-562 and B16-F10) in vitro. sPLA2s inhibited collagen-induced platelet aggregation: VBBPLA2 IC50 = 0.054, VLPLA2 IC50 = 0.072, NNOPLA2 IC50 = 0.814 μM. p-Bromophenacylbromide-inhibited sPLA2 had no inhibitory action on platelets. 36.17 μM VBBPLA2 completely inhibited the growth of gram-positive Bacillus subtilis whereas no growth inhibition was observed towards gram-negative Escherichia coli. The inhibitory action of sPLA2s (~0.7 μM and ~7 μM) towards cancer cells depended on both venom and cell type. VBBPLA2 (7.2 μM) inhibited significantly the viability of K-562 cells and the cell death appeared apoptotic. The sPLA2s exhibited no inhibitory effect towards LNCaP cells and some effect (8%–20%) towards other cells. Thus, already sub-μM concentrations of sPLA2s inhibited collagen-induced platelet aggregation and from the current suite of studied svPLA2s and test cells, VBBPLA2 was the most growth inhibitory towards Bacillus subtilis and K-562 cells.

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“…2 Snake venom also contains phospholipase A2, which produces local inflammation and tissue necrosis and hemorrhagin, which causes endothelial damage. 10 Also, neurotoxins and coagulants can release inflammatory mediators causing local inflammation and stasis of bile. Another possible theory postulated is change in the chemical composition of bile probably caused by phospholipase A2, which converts phospholipids in bile into toxic fatty acids and lysolecithin.…”

Section: Discussionmentioning

confidence: 99%

Acute acalculous cholecystitis as a presenting feature of neurotoxic snake bite: a case report and review of literature

2018

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Snakebite is classified by the WHO as a neglected tropical disease. Neurotoxic snake bites pose a great diagnostic challenge to the physicians in tropical and subtropical regions of the world. There is considerable variation between individual patients in the clinical manifestations following envenoming by neurotoxic snake bite. Here authors describe a rare case who presented to emergency department as acute acalculous cholecystitis following unknown neurotoxic snake bite. This acalculous cholecystitis resolved post administration of anti-snake venom. The possibility of this rare but potentially fatal complication needs to be considered in patients with snake bite and abdominal symptoms so that prompt management can prevent mortality in such patients.

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Purification and characterization of a platelet aggregation inhibitor acidic phospholipase A2 from Indian saw-scaled viper (Echis carinatus) venom

Cited by 34 publications

References 29 publications

Proteins from Mucuna pruriens and Enzymes fromEchis carinatus Venom

Proteins from Mucuna pruriens and Enzymes fromEchis carinatus Venom

Interactions of PLA2-s from Vipera lebetina, Vipera berus berus and Naja naja oxiana Venom with Platelets, Bacterial and Cancer Cells

Acute acalculous cholecystitis as a presenting feature of neurotoxic snake bite: a case report and review of literature

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