Proteins from Mucuna pruriens and Enzymes fromEchis carinatus Venom

Abstract

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...

Deturbidization of Petroleum-Produced Water by Novel Mucuna Seed: Characterization, Kinetics, and Statistical Analyses

“…It could be observed that the best performances were obtained at acidic condition. These results of good performance in acidic medium are supported by previous reports [6]. Figure 8 specifically revealed that the maximum efficiency of 96.41% was obtained at pH 4.…”

“…Mucuna is of an annual twinning tropical plant, which contains thickening seed bean that is composed of soluble proteins (20-30%), lipids, fibers, and minerals [6]. Results obtained from the preliminary application of novel Mucuna seed coagulant (MSC) in the treatment of coal washery effluent indicated its prospects for potential utilization in the treatment of produced water (PW), which constitutes aquatic pollutant in Nigeria's Niger Delta region [7,8].…”

Purification and characterization of a glycoprotein inhibitor of toxic phospholipase from Withania somnifera

“…The activity may be attributed to the presence of enzymatic inhibitors, chemical inactivators, or immunomodulators present in the plant isolates [11]. Guerranti et al [12] demonstrated that the presence of proteins from seed extract of Mucuna pruiens protected mice against the toxic effects of Echis carinatus venom. It does so by an immunological mechanism based on a series of specific epitopes common to some vegetal and venom proteins.…”

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“…The mechanisms of the protective effects exerted by M. pruriens seed aqueous extract (MPE) were investigated in detail in a study involving the effects of echis carinatus venom (EV). [15] In vivo experiments on mice showed that protection against the poison is evident at 24 h (short term) and 1 month (long term) after injection of MPE. [16] MPE protects mice against the toxic effects of EV through an immune mechanism.…”

“…M. pruriens plant has many activities which need to elaborate for future cure in Ayurvedic tradition. [15] Antidiabetic Plant seed Ethanol/water (1:1) Cyclitols, oligosaccharides [21] Antioxidant Whole plant Methanol Phenols, tannins [22] Neuroprotective Plant seed, whole plant Ethanol/water (1:1) L-DOPA, amino acids, alkaloids n-propanol Isoquinolines, alkaloids [25] Antimicrobial Plant leave Methanol Tannins, alkaloids, L-DOPA [27] …”