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N-terminal-sequences and CD spectrum of CC-PLA2-1 and CC-PLA2-2 (A) N-terminal sequence homology between CC-PLA2-1, CC-PLA2-2, PLA2_CERCE from Pakistanian Cerastes cerastes, Myotoxin II from Bothrops nummifer and BthA-I-PLA2 from Bothrops jararacussu of group II phospholipases A2 from viperid snake venom. (B) The CD spectra of CC-PLA2-1 (d) and CC-PLA2-2 (--) were measured as described in the Materials and Methods. Data were expressed in variation of molar amino acid residue absorption coefficient (D3).
Source publication
Two non-toxic PLA2s were purified to homogeneity from Cerastes cerastes tunisian snake venom. The purification process employed gel filtration on Sephadex G-75 followed by C18 reverse phase high-pressure liquid chromatography. These two acidic enzymes, namely CC-PLA2-1 and CC-PLA2-2, have a molecular weight of 13737.52 and 13705.63 Da, respectively...
Contexts in source publication
Context 1
... N-terminal amino acid sequences (50 amino acids) of CC-PLA2-1 and CC-PLA2-2 are very similar (90% of identity). Indeed, (Fig. ...
Context 2
... showing sequence similarity with CC-PLA2-1 and CC-PLA2-2 were identified in the non-redundant database with BLAST2 program ( Altschul et al., 1997). Best hits were group IIA PLA2s. These were aligned and sequence similarities were determined using Bioedit program (www.mbio.ncsu.edu/BioEdit/bioedit.html) ( Fig. ...
Context 3
... illustrated in Fig. 2B, the CD spectrum showed a similar structure for both purified PLA2 and indicated a high content in helical structures. The CD spectra show two negative bands at 222 and 205 nm and a positive band at 190 nm that are typical of a helix. However, the high intensity of the a helix signal can hide contribution of other secondary structures ...
Context 4
... show herein the presence in the C. cerastes venom of two PLA2, termed CC-PLA2-1 and CC-PLA2-2. N-terminal sequences of both CC-PLA2s showed a high level of identity with Group II PLA2s (w64-88%), especially with PLA2 CERCE, isolated from Pakistanian C. cerastes (Siddiqui et al., 1991) (Fig. 2A). This data may be an agreement with venom composition variation from different geographical area and may reveal intra-species variability (Chippaux et al., ...
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Three-step chromatography and proteomic analysis have been used to purify and characterize a new basic phospholipase A2 named CC2-PLA2 from the venom of Cerastes cerastes. This phospholipase A2 has been isolated to an extent of about 50-folds and its molecular weight was estimated at 13,534 Da. For CC2-PLA2 identification and LC-MALDI-MS/MS analysi...
L-ascorbic acid upon condensation with palmitic acid in the presence of sulphuric acid results in L-ascorbic acid-6-palmitate (AP). The effect of L-ascorbic acid derivative, AP on the pharmacological activities of purified basic multi-toxic PLA2 enzyme, VRV-PL-VIIIa from Vipera russelli snake venom along with in vitro activities is described. AP in...
Citations
... The venom of the Macrovipera lebetina is an amalgam of different proteins, including proteases, phosphodiesterases, phosphatases, phospholipase A2, disintegrins, and C-type lectin-like proteins [8][9][10][11][12][13][14]. Among the C-type lectin-like proteins from Macrovipera lebetina, lebectin and lebecin, showed a powerful inhibition of platelet aggregation, cell adhesion, migration, invasion, and proliferation by inhibiting the 5 1 and v integrins without inhibition of the collagen 2 1 receptor [15,16]. ...
... Another study showed that Asp-49 PLA 2 from the venom of B. jararacussu has anti-tumor and antimetastatic effects on the same cell line suggesting that this protein may be used for drug development for the treatment of breast cancer [68]. Additionally, CC-PLA2-1 and CC-PLA2-2, isolated from the venom of Cerastes cerastes (Horned desert viper), have been shown to effectively inhibit tumor cell adhesion and migration by interfering with α5β1 and αv integrins, resulting in angiogenesis blocking [69]. ...
Snake venom is a rich source of bioactive molecules that hold great promise for therapeutic applications. These molecules can be broadly classified into enzymes and non-enzymes, each showcasing unique medicinal properties. Noteworthy compounds such as Bradykinin Potentiating Peptides (BPP) and Three-Finger Toxins (3FTx) are showing therapeutic potential in areas like cardiovascular diseases (CVDs) and pain-relief. Meanwhile, components like snake venom metalloproteinases (SVMP), L-amino acid oxidases (LAAO), and Phospholipase A2s (PLA2) are paving new ways in oncology treatments. The full medicinal scope of these toxins is still emerging. In this review, we discuss drugs derived from snake venoms that address CVDs, cancer, diabetes, strokes, and pain. Further, we outline the toxin distribution across 130 snake species, categorized by their genus within the Crotalidae, Viperidae, and Elapidae families. Conclusively, we spotlight the potential of Deep Learning (DL) in discovering groundbreaking drug prospects from these toxins.
... It has been proven that numerous phospholipases, including basic and acidic PLA 2 s, as well as synthetic peptides derived from PLA 2 homologs, have been shown to have anticancer and antiangiogenic properties [92,93]. Two phospholipases A 2 from the venom of Cerastes cerastes, CC-PLA2-1 and CC-PLA2-2, were recently isolated and characterized [94]. ...
Snake bite is a neglected disease that affects millions of people worldwide. WHO reported approximately 5 million people are bitten by various species of snakes each year, resulting in nearly 1 million deaths and an additional three times cases of permanent disability. Snakes utilize the venom mainly for immobilization and digestion of their prey. Snake venom is a composition of proteins and enzymes which is responsible for its diverse pharmacological action. Snake venom phospholipase A2 (SvPLA2) is an enzyme that is present in every snake species in different quantities and is known to produce remarkable functional diversity and pharmacological action like inflammation, necrosis, myonecrosis, hemorrhage, etc. Arachidonic acid, a precursor to eicosanoids, such as prostaglandins and leukotrienes, is released when SvPLA2 catalyzes the hydrolysis of the sn-2 positions of membrane glycerophospholipids, which is responsible for its actions. Polyvalent antivenom produced from horses or lambs is the standard treatment for snake envenomation, although it has many drawbacks. Traditional medical practitioners treat snake bites using plants and other remedies as a sustainable alternative. More than 500 plant species from more than 100 families reported having venom-neutralizing abilities. Plant-derived secondary metabolites have the ability to reduce the venom's adverse consequences. Numerous studies have documented the ability of plant chemicals to inhibit the enzymes found in snake venom. Research in recent years has shown that various small molecules, such as varespladib and methyl varespladib, effectively inhibit the PLA2 toxin. In the present article, we have overviewed the knowledge of snake venom phospholipase A2, its classification, and the mechanism involved in the pathophysiology of cytotoxicity, myonecrosis, anticoagulation, and inflammation clinical application and inhibitors of SvPLA2, along with the list of studies carried out to evaluate the potency of small molecules like varespladib and secondary metabolites from the traditional medicine for their anti-PLA2 effect.
... The same stability profile was also observed with Wa-PLA 2 -II, for which total activity was maintained between pH 6 and 10, and less than 40% of the original activity was lost at pH 12 or 4 [7]. Cc-PLA 2 purified from the venom of Tunisian Cerastes cerastes were also stable at pH from 3 to 9 [37]. Yuan et al. explained the stability at extreme pH through an RMN study of porcine PLA 2 . ...
Secreted phospholipases A2 are snake-venom proteins with many biological activities, notably anti-tumor activity. Phospholipases from the same snake type but different geographical locations have shown similar biochemical and biological activities with minor differences in protein sequences. Thus, the discovery of a new phospholipase A2 with unique characteristics identified in a previously studied venom could suggest the origins of these differences. Here, a new Group II secreted phospholipase A2 (Cc-PLA2-II) from the snake venom of Saudi Cerastes cerastes gasperetti was isolated and characterized. The purified enzyme had a molecular weight of 13.945 kDa and showed high specific activity on emulsified phosphatidylcholine of 1560 U/mg at pH 9.5 and 50 °C with strict calcium dependence. Interestingly, stability in extreme pH and high temperatures was observed after enzyme incubation at several pH levels and temperatures. Moreover, a significant dose-dependent cytotoxic anti-tumor effect against six human cancer cell lines was observed with concentrations of Cc-PLA2 ranging from 2.5 to 8 µM. No cytotoxic effect on normal human umbilical-vein endothelial cells was noted. These results suggest that Cc-PLA2-II potentially has angiogenic activity of besides cytotoxicity as part of its anti-tumor mechanism. This study justifies the inclusion of this enzyme in many applications for anticancer drug development.
... Despite the growing number of studies using animal toxins for therapeutic and anti-tumor purposes in recent decades, some snake venom proteins and their respective compounds are still poorly studied with regard to the melanoma model. In our systematic review, we found these protein families that have been little explored so far: Serinoproteases [58]; PLA 2 : CC-PLA 2 -1 and CC-PLA 2 -2, two glycosylated, acidic enzymes, non-toxic PLA 2 s [162]; Cathelicidin-like polypeptide: Cathelicidin-BF (BF-30) is a cathelicidin-like polypeptide [68] ; Cystatin: Cystatin (sv-cystatin) [67]; Cytotoxin-II (rCTII) [19]; Crotalase, thrombin-like enzyme [189]. ...
Despite advances in treating patients with melanoma, there are still many treatment challenges to overcome. Studies with snake venom-derived proteins/peptides describe their binding potential, and inhibition of some proliferative mechanisms in melanoma. The combined use of these compounds with current therapies could be the strategic gap that will help us discover more effective treatments for melanoma. The present study aimed to carry out a systematic review identifying snake venom proteins and peptides described in the literature with antitumor, antimetastatic, or antiangiogenic effects on melanoma and determine the mechanisms of action that lead to these anti-tumor effects. Snake venoms contain proteins and peptides which are antiaggregant, antimetastatic, and antiangiogenic. The in vivo results are encouraging, considering the reduction of metastases and tumor size after treatment. In addition to these results, it was reported that these venom compounds could act in combination with chemotherapeutics (Acurhagin-C; Macrovipecetin), sensitizing and preparing tumor cells for treatment. There is a consensus that snake venom is a promising strategy for the improvement of antimelanoma therapies, but it has been little explored in the current context, combined with inhibitors, immunotherapy or tumor microenvironment, for example. We suggest Lebein as a candidate for combination therapy with BRAF inhibitors.
... The involvement of SVPLA 2 in hemorrhage and dermonecrosis was also highlighted for Daboia russelii venom [61]. It was reported that several SVPLA 2 isolated from C. cerastes venom are able to inhibit platelet aggregation and coagulation [49,62]. ...
Envenomation by Cerastes cerastes often results in local dermonecrotic lesions. While immunotherapy is effective in reversing systemic symptoms, this strategy remains deficient in counteracting the extended dermonecrosis induced from the bite site. In this study, the therapeutic effect of pharmacological drugs on the dermonecrotic activity of the venom was investigated. Venom administration caused a marked dermonecrotic lesion with increased levels of oxidative stress biomarkers (MPO, EPO, NO, H2O2, MDA, protein carbonyl, and thiol levels). Antioxidant capacity was decreased, as evidenced by reduced catalase, glutathione, and selenium levels. Histopathological analysis of skin biopsies revealed necrotic lesions accompanied by hemorrhage and epidermis thickening. The efficiency of cyproheptadine (C), dexamethasone (D), and tetracycline (T), as a monotherapy or in association, were evaluated on the dermonecrotic activity of the venom. Most of the treatments (CD, CT, DT, and CDT) largely reduced tissue necrosis to, respectively, 84.29, 87.83, 83.77, and 82.71% and significantly decreased MPO and EPO activities and NO, H2O2, MDA, and protein carbonyl levels in skin tissue homogenates. CT and CDT associations significantly increased the antioxidant status as indicated by enhanced catalase, glutathione, and selenium levels. The second challenge of the pharmacological associations was more effective in improving the oxidative/antioxidative balance. Skin tissue sections from treated animals with CT or CDT revealed tissue structure close to that observed in control animals. Therefore, the synergistic action of all tested drugs on the major pathways of inflammation (phospholipases A2, metalloproteinases, and histamine) seems to be efficient to neutralize the necrotic activity of the venom.
... derived from snake venoms possess antitumor activity owing to their inhibitory action against several tumor cells (Rudrammaji and Gowda, 1998;Kang et al., 2000;Rodrigues et al., 2009). The cell-cell interactions (Kang et al., 2000), along with cell adhesion and migration functions (Zouari-Kessentini et al., 2009), are not necessarily dependent on their catalytic activity (Stabeli et al., 2006). Crotoxin (CTX) is the most abundant toxin (nearly 60% of the total venom) in Crotalus durissus terrificus (Laurenti, 1768) venom (CdtV). ...
Antitumor property of Crotoxin (CTX), the major toxin from Crotalus durissus terrificus snake venom, has been demonstrated in experimental animal models and clinical trials. However, the direct action of this toxin on the significant events involved in neovascularization, which are essential for tumor growth and survival, has not been confirmed. This study investigated the effects of CTX on the key parameters of neovascularization in two- and three-dimensional culture models. Murine endothelial cell lines derived from thymus hemangioma (t.End.1) were treated at different concentrations of CTX (6.25–200 nM). Endothelial cell proliferation, cell adhesion, and actin cytoskeletal dynamics on laminin (10 µg/ml), type I collagen (10 µg/ml), and fibronectin (3 µg/ml) were evaluated along with the endothelial cell migration and formation of capillary-like tubes in 3D Matrigel. CTX concentration of 50 nM inhibited tube formation on 3D Matrigel and impaired cell adhesion, proliferation, and migration under both culture medium and tumor-conditioned medium. These actions were not accountable for the loss of cell viability. Inhibition of cell adhesion to different extracellular matrix components was related to the reduction of αv and α2 integrin distribution and cytoskeletal actin polymerization (F-actin), accompanied by inhibition of focal adhesion kinase (FAK), Rac1 (GTPase) signaling proteins, and actin-related protein 2/3 (Arp 2/3) complex. This study proved that CTX inhibits the major events involved in angiogenesis, particularly against tumor stimuli, highlighting the importance of the anti-angiogenic action of CTX in inhibition of tumor progression.
... In-vitro researches have demonstrated that the pharmacological effects of snakes' poison are dose-dependent. The studied species were namely Macrovipera lebetina and Cerastes cerastes, which are major species in our region [7,25,26]. The poison dose is frequently correlated with snake size and/or bites number. ...
Objective: To identify the early predictors of severe envenomation in the southern region of Tunisia.
Methods: It was a retrospective monocentric study including consecutive patients admitted to the emergency department for snakebite envenomation. Snakebite envenomation was defined by a history of snakebite. Predictors of severe envenomation were determined by univariate and multivariate analyses.
Results: Our sample consisted of 109 patients aged 30 (20-44) years with a 1.1:1 sex-ratio (56 males and 53 females). During the 24-hour surveillance period, 25 patients developed severe envenomation (22.9%). The in-hospital mortality rate was 4.6% (n=5). The independent predictors of severe snakebite envenomation were leucocyte count over 11 550/mm3 (OR: 18.7, 95% CI: 3.3-107.8), creatine kinase over 155 IU/L (OR: 6.16, 95% CI: 1.1-35.6), and/or tourniquet before arrival to the ED (OR: 32.14, 95% CI: 3.5-295.9).
Conclusions: This study emphasizes the importance of early evaluation of snakebite envenomation. Further studies are required to approve a severity scale proper to snakebite envenomation in Tunisia.
... Interestingly, SPLA 2 -IB used as a non-glycosylated control, WaPLA 2 -II was found to be glycosylated with a carbohydrate content of 2.5% compared with 2% for WaPLA 2 -I [40]. This finding is in accordance with that obtained by Zouari-Kessentini et al. who reported the first glycosylated PLA2 enzyme purified from venom of the Tunisian snake, Cerastes cerastes [50]. Moreover, a glycosylated form of the marine SPLA2-IIA was also identified with a carbohydrate content of 2.5% (data not shown) [8]. ...
... The NH 2 -terminal amino acid sequence of pure WaPLA 2 -II was determined (NLQFGMIKLTTGKPEALSYNAYGCYCGWGGQGKPQDATDHCCFVHDCC) and, as it is shown in Figure 1D, it exhibited a high similarity (≥83%) with the available sequences of secreted PLA2 G II isolated from the Cerastes cerastes [22], Echis carinatus, Vipera renardi, and Atheris nitschei snake venoms [22,50,51] ( Figure 1D). A relatively low similarity was obtained with sPLA2 G I (≤55%) from Bungarus fasciatus venom [49]. ...
... However, further deep investigations (such as the determination of the entire sequence) are required to confirm that WaPLA 2 -II belongs to G II sPLA2. It is worthy to notice that G II svPLA2s have always been present in viperid and crotalid snakes, whereas G I svPLA2s are found in elapid and hydrophid snakes [50]. To our knowledge, we reported here for the first time, the purification to homogeneity of this svPLA2 G II from the snake family of Elapidae, from Saudi Arabia. ...
Many venomous species, especially snakes, contain a variety of secreted phospholipases A2 that contribute to venom toxicity and prey digestion. We characterized a novel highly toxic phospholipase A2 of group II, WaPLA2-II, from the snake venom of Saudi Walterinnesia aegyptia (W. aegyptia). The enzyme was purified using a reverse phase C18 column. It is a monomeric protein with a molecular weight of approximately 14 kDa and an NH2-terminal amino acid sequence exhibiting similarity to the PLA2 group II enzymes. WaPLA2-II, which contains 2.5% (w/w) glycosylation, reached a maximal specific activity of 1250 U/mg at pH 9.5 and 55 °C in the presence of Ca2+ and bile salts. WaPLA2-II was also highly stable over a large pH and temperature range. A strong correlation between antimicrobial and indirect hemolytic activities of WaPLA2 was observed. Additionally, WaPLA2-II was found to be significantly cytotoxic only on cancerous cells. However, chemical modification with para-Bromophenacyl bromide (p-BPB) inhibited WaPLA2-II enzymatic activity without affecting its antitumor effect, suggesting the presence of a separate ‘pharmacological site’ in snake venom phospholipase A2 via its receptor binding affinity. This enzyme is a candidate for applications including the treatment of phospholipid-rich industrial effluents and for the food production industry. Furthermore, it may represent a new therapeutic lead molecule for treating cancer and microbial infections.
... Da, respectively, were purified (Fig. 4). These enzymes strongly inhibited coagulation and exhibited a marked inhibitory effect on platelet aggregation induced by ADP and arachidonic acid [48]. Interestingly, CC-PLA2-1 and CC-PLA2-2 impaired in a dose-dependent manner adhesion and migration of HT1080 fibrosarcoma and IGR39 melanoma cells to fibrinogen and fibronectin [48]. ...
... These enzymes strongly inhibited coagulation and exhibited a marked inhibitory effect on platelet aggregation induced by ADP and arachidonic acid [48]. Interestingly, CC-PLA2-1 and CC-PLA2-2 impaired in a dose-dependent manner adhesion and migration of HT1080 fibrosarcoma and IGR39 melanoma cells to fibrinogen and fibronectin [48]. These enzymes also inhibited HBMEC cell adhesion and migration to fibrinogen and fibronectin. ...
... Extensive studies conducted in vitro, ex vivo, or in vivo, have shown that some of these compounds can interfere with hemostasis or display anti-or pro-aggregating platelet properties [15,33,56,57]. Some others have anti-tumor effect via several pathways [12,20,27,48,51]. Other molecules can even, by acting on transmembrane receptors, prevent ischemic or neovascular disorders in a broad range of diseases [16,35,36]. ...
The venoms of Tunisian wildlife snakes are complex mixtures containing proteins/peptides and non-protein molecules. Proteins and peptides are the most abundant compounds responsible for the biological effects of venoms. Snake venoms proteins have enzymatic or non-enzymatic activities, which are grouped into different families including C-type lectin proteins, disintegrins (long, medium and short disintegrins), Kunitz-type serine protease inhibitors, natriuretic-like peptides, vascular endothelial growth factor-related proteins, L-amino acid oxidases, phospholipases A2 and serine proteinases. With technological advancements, the toxic effects of venoms were turned into potential benefits for clinical diagnosis, basic research and development of new research tools and drugs of potential clinical use. Our research team has shown that Macrovipera lebetina and Cerastes cerastes venom components of Tunisian wildlife snakes had great potential for the development of new drugs for the treatment of cancer, angiogenesis disorders or cardiovascular diseases. This review is an overview on snake venom proteins from Macrovipera lebetina and Cerastes cerastes and their biochemical, pharmacological and molecular characterization and their importance as protein resources with therapeutic potential.
