scholarly journals Snake Venom Proteomics of Samar Cobra (Naja samarensis) from the Southern Philippines: Short Alpha-Neurotoxins as the Dominant Lethal Component Weakly Cross-Neutralized by the Philippine Cobra Antivenom

2021 ◽  
Vol 12 ◽  
Author(s):  
Praneetha Palasuberniam ◽  
Yi Wei Chan ◽  
Kae Yi Tan ◽  
Choo Hock Tan
Keyword(s):  
Snake Venom ◽  
High Performance ◽  
Lethal Dose ◽  
Secretory Proteins ◽  
Venomous Snake ◽  
Neutralization Activity ◽  
Medical Importance ◽  
Species Specific ◽  
Venom Proteins

The Samar Cobra, Naja samarensis, is endemic to the southern Philippines and is a WHO-listed Category 1 venomous snake species of medical importance. Envenomation caused by N. samarensis results in neurotoxicity, while there is no species-specific antivenom available for its treatment. The composition and neutralization of N. samarensis venom remain largely unknown to date. This study thus aimed to investigate the venom proteome of N. samarensis for a comprehensive profiling of the venom composition, and to examine the immunorecognition as well as neutralization of its toxins by a hetero-specific antivenom. Applying C18 reverse-phase high-performance liquid chromatography (RP-HPLC) and tandem mass spectrometry (LC-MS/MS), three-finger toxins (3FTx) were shown to dominate the venom proteome by 90.48% of total venom proteins. Other proteins in the venom comprised snake venom metalloproteinases, phospholipases A2, cysteine-rich secretory proteins, venom nerve growth factors, L-amino acid oxidases and vespryn, which were present at much lower abundances. Among all, short-chain alpha-neurotoxins (SαNTX) were the most highly expressed toxin within 3FTx family, constituting 65.87% of the total venom proteins. The SαNTX is the sole neurotoxic component of the venom and has an intravenous median lethal dose (LD50) of 0.18 μg/g in mice. The high abundance and low LD50 support the potent lethal activity of N. samarensis venom. The hetero-specific antivenom, Philippine Cobra Antivenom (PCAV, raised against Naja philippinensis) were immunoreactive toward the venom and its protein fractions, including the principal SαNTX. In efficacy study, PCAV was able to cross-neutralize the lethality of SαNTX albeit the effect was weak with a low potency of 0.20 mg/ml (defined as the amount of toxin completely neutralized per milliliter of the antivenom). With a volume of 5 ml, each vial of PCAV may cross-neutralize approximately 1 mg of the toxin in vivo. The findings support the potential para-specific use of PCAV in treating envenomation caused by N. samarensis while underscoring the need to improve the potency of its neutralization activity, especially against the highly lethal alpha-neurotoxins.

scholarly journals ASSESSMENT OF SNAKE ANTIVENOM ACTIVITY OF TAMARINDUS INDICA SEED COAT EXTRACT

2021 ◽  
Vol 9 (09) ◽  
pp. 489-497
Author(s):  
Priyanka D. Mundhe ◽  
◽  
Balasaheb S. Pawade ◽  
Indrasen G. Waykar ◽  
Innus K. Shaikh ◽  
...  
Keyword(s):  
Snake Venom ◽  
Seed Coat ◽  
Rural Areas ◽  
Proteolytic Enzymes ◽  
Lethal Dose ◽  
Medical Emergency ◽  
Dependent Manner ◽  
Tamarindus Indica

Snakebite is a life-threatening medical emergency, and globally responsible for millions of deaths. In snakebites accidents only deaths are not a concern, it leads to more morbidities. Due to scanty healthcare facilities in rural areas of India, many people seek alternative treatment available in ethnic practices. Tamarindus Indica (TI) plant is rich in medicinal value and used to treat many diseases including snakebite treatment traditionally. In view of this TI seed coat extract (TISCE) was evaluated for antivenom activity. The phytochemical screening of TISCE was performed to understand its chemical composition. TISCE was evaluated for antivenom activity against Indian cobra venom (ICV), common krait venom (CKV), Russells viper venom (RVV), and saw-scaled viper venom (SCV) for phospholipase A2 (PLA-2), haemorrhagic in vitro and in vivo, procoagulant, proteolytic activity, and lethality studies. TISCE majorly contains saponins, glycosides, alkaloids, and phenolic compounds. Minimum indirect haemorrhagic dose (MIHD) observed for ICV (12.5 µg), CKV (5.0 µg),RVV (10.0 µg), and SVV (12.5 µg). TISCE inhibits the procoagulant activity of all venoms at a concentration of 18.0 µg. It also shows the neutralization of proteolytic enzymes of venom in a dose-dependent manner. A pre-incubated mixture containing five lethal dose 50 (LD50) of venom and TISCE was injected intravenously, all mice survived as venom neutralized by TISCE. The present study demonstrates the ability of TISCE to neutralize snake venom using suitable in vivo and in vitro methods. Further studies required to unravelling the specific active chemical constituent of TISCE that may used as novel alternative snakebite treatment. TISCE was able to prolong the deaths during the simulation study and may be used in the topical pharmaceutical formulation that will reduce local venom reactions causing much morbidity, which will collectively with Anti-snake venom (ASV), used to treat envenomed patients more effectively.

scholarly journals Cysteine-Rich Secretory Proteins (CRISPs) from Venomous Snakes: An Overview of the Functional Diversity in a Large and Underappreciated Superfamily

Toxins ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 175 ◽  
Author(s):  
Takashi Tadokoro ◽  
Cassandra M. Modahl ◽  
Katsumi Maenaka ◽  
Narumi Aoki-Shioi
Keyword(s):  
Snake Venom ◽  
Inflammatory Responses ◽  
Neutrophil Infiltration ◽  
Secretory Proteins ◽  
Sequence Alignments ◽  
Functional Sites ◽  
Accelerated Evolution ◽  
Evolutionarily Conserved ◽  
Pathogenesis Related ◽  
Venom Proteins

The CAP protein superfamily (Cysteine-rich secretory proteins (CRISPs), Antigen 5 (Ag5), and Pathogenesis-related 1 (PR-1) proteins) is widely distributed, but for toxinologists, snake venom CRISPs are the most familiar members. Although CRISPs are found in the majority of venoms, very few of these proteins have been functionally characterized, but those that have been exhibit diverse activities. Snake venom CRISPs (svCRISPs) inhibit ion channels and the growth of new blood vessels (angiogenesis). They also increase vascular permeability and promote inflammatory responses (leukocyte and neutrophil infiltration). Interestingly, CRISPs in lamprey buccal gland secretions also manifest some of these activities, suggesting an evolutionarily conserved function. As we strive to better understand the functions that CRISPs serve in venoms, it is worth considering the broad range of CRISP physiological activities throughout the animal kingdom. In this review, we summarize those activities, known crystal structures and sequence alignments, and we discuss predicted functional sites. CRISPs may not be lethal or major components of venoms, but given their almost ubiquitous occurrence in venoms and the accelerated evolution of svCRISP genes, these venom proteins are likely to have functions worth investigating.

scholarly journals Venomics of Trimeresurus (Popeia) nebularis, the Cameron Highlands Pit Viper from Malaysia: Insights into Venom Proteome, Toxicity and Neutralization of Antivenom

Toxins ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 95 ◽  
Author(s):  
Choo Tan ◽  
Kae Tan ◽  
Tzu Ng ◽  
Evan Quah ◽  
Ahmad Ismail ◽  
...  
Keyword(s):  
Snake Venom ◽  
Serine Proteases ◽  
Cross Reactivity ◽  
Secretory Proteins ◽  
Proteomic Profiling ◽  
Phospholipases A2 ◽  
Central Highland ◽  
Liquid Chromatography Tandem Mass ◽  
Cameron Highlands ◽  
Venom Proteins

Trimeresurus nebularis is a montane pit viper that causes bites and envenomation to various communities in the central highland region of Malaysia, in particular Cameron’s Highlands. To unravel the venom composition of this species, the venom proteins were digested by trypsin and subjected to nano-liquid chromatography-tandem mass spectrometry (LC-MS/MS) for proteomic profiling. Snake venom metalloproteinases (SVMP) dominated the venom proteome by 48.42% of total venom proteins, with a characteristic distribution of P-III: P-II classes in a ratio of 2:1, while P-I class was undetected. Snaclecs constituted the second most venomous protein family (19.43%), followed by snake venom serine proteases (SVSP, 14.27%), phospholipases A2 (5.40%), disintegrins (5.26%) and minor proteins including cysteine-rich secretory proteins, L-amino acid oxidases, phosphodiesterases, 5′-nucleotidases. The venomic profile correlates with local (painful progressive edema) and systemic (hemorrhage, coagulopathy, thrombocytopenia) manifestation of T. nebularis envenoming. As specific antivenom is unavailable for T. nebularis, the hetero-specific Thai Green Pit viper Monovalent Antivenom (GPVAV) was examined for immunological cross-reactivity. GPVAV exhibited good immunoreactivity to T. nebularis venom and the antivenom effectively cross-neutralized the hemotoxic and lethal effects of T. nebularis (lethality neutralizing potency = 1.6 mg venom per mL antivenom). The findings supported GPVAV use in treating T. nebularis envenoming.

scholarly journals Venom Proteome of Spine-Bellied Sea Snake (Hydrophis curtus) from Penang, Malaysia: Toxicity Correlation, Immunoprofiling and Cross-Neutralization by Sea Snake Antivenom

Toxins ◽  
2018 ◽  
Vol 11 (1) ◽  
pp. 3 ◽  
Author(s):  
Choo Tan ◽  
Kae Tan ◽  
Tzu Ng ◽  
Si Sim ◽  
Nget Tan
Keyword(s):  
Liquid Chromatography ◽  
High Performance ◽  
Secretory Protein ◽  
Major Protein ◽  
Sea Snake ◽  
Main Driver ◽  
Liquid Chromatography Tandem Mass ◽  
Cross Neutralization ◽  
Venom Proteins

The venom proteome of Hydrophis curtus (synonym: Lapemis hardwickii) from Penang, Malaysia was investigated with nano-electrospray ionization-liquid chromatography tandem mass spectrometry (ESI-LCMS/MS) of the reverse-phase high-performance liquid chromatography (HPLC) venom fractions. Thirty distinct protein forms were identified as toxins from ten families. The three major protein families were phospholipase A2 (PLA2, 62.0% of total venom proteins), three-finger toxin (3FTX, 26.33%) and cysteine-rich secretory protein (CRiSP, 9.00%). PLA2 comprises diverse homologues (11 forms), predominantly the acidic subtypes (48.26%). 3FTX composed of one short alpha-neurotoxin (SNTX, 22.89%) and four long alpha-neurotoxins (LNTX, 3.44%). Both SNTX and LNTX were lethal in mice (intravenous LD50 = 0.10 and 0.24 μg/g, respectively) but the PLA2 were non-lethal (LD50 >1 μg/g). The more abundant and toxic SNTX appeared to be the main driver of venom lethality (holovenom LD50 = 0.20 μg/g). The heterologous Sea Snake Antivenom (SSAV, Australia) effectively cross-neutralized the venom (normalized potency = 9.35 mg venom neutralized per g antivenom) and the two neurotoxins in vivo, with the LNTX being neutralized more effectively (normalized potency = 3.5 mg toxin/g antivenom) than SNTX (normalized potency = 1.57 mg/g). SSAV immunorecognition was strong toward PLA2 but moderate-to-weak toward the alpha-neurotoxins, indicating that neutralization of the alpha-neurotoxins should be further improved.

scholarly journals Gene Cloning of Rat and Mouse Platelet Glycoprotein V: Identification of Megakaryocyte-Specific Promoters and Demonstration of Functional Thrombin Cleavage

Blood ◽  
1997 ◽  
Vol 89 (9) ◽  
pp. 3253-3262 ◽  
Author(s):  
Catherine Ravanat ◽  
Martine Morales ◽  
David O. Azorsa ◽  
Sylvie Moog ◽  
Simone Schuhler ◽  
...  
Keyword(s):  
High Performance ◽  
Surface Protein ◽  
Platelet Glycoprotein ◽  
Recognition Sequence ◽  
Thrombin Cleavage ◽  
Human Thrombin ◽  
Recognition Motifs ◽  
Species Specific ◽  
Major Surface

Abstract Platelet glycoprotein (GP) V is a major surface protein cleaved during thrombin-induced platelet activation. GPV associates noncovalently with the GPIb-IX complex to form GPIb-V–IX, a receptor for von Willebrand factor and thrombin. We describe the cloning of the genes coding for rat and mouse GPV and compare them with the human gene. The two rodent genes have a similar structure and resemble the human GPV gene with a coding sequence (≈1,700 nucleotides) entirely contained in one exon and a single intron (≈900 nucleotides) in the 5′ untranslated region. Both genes have megakaryocyte-type promoters with conserved tandem Ets and GATA recognition motifs and lack a TATA box. The mature rat and mouse proteins comprise 551 amino acids, have 70% sequence identity, and contain an additional 8–amino acid intracellular segment as compared with the human protein. As in human GPV, there is an NH2 -terminal leucine-rich region of 15 repeats and a thrombin cleavage recognition sequence. Whereas the rat and human thrombin cleavage sites are similar, the mouse cleavage site resembles that of the human thrombin receptor. Functionality of these sites was demonstrated by thrombin cleavage of synthetic peptides and analysis by high-performance liquid chromatography (HPLC) or mass spectrometry. Cleavage of native rat GPV was confirmed by means of a polyclonal antibody directed against the new NH2 -terminal peptide exposed after thrombin cleavage. This antibody specifically recognized thrombin-activated rat platelets by fluorescence-activated cell sorting (FACS) analysis. In addition, we raised monoclonal antibodies specific for rat GPV (88 kD), which recognized the NH2 -terminal soluble fragment (70 kD) liberated after thrombin cleavage. Knowledge of these rodent GPV genes and availability of species-specific peptides and antibodies will be essential to further studies aiming to define the exact in vivo function of platelet GPV using animal models of thrombosis and gene inactivation experiments.

scholarly journals Snake Venom Proteomics, Immunoreactivity and Toxicity Neutralization Studies for the Asiatic Mountain Pit Vipers, Ovophis convictus, Ovophis tonkinensis, and Hime Habu, Ovophis okinavensis

Toxins ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 514
Author(s):  
Choo Hock Tan ◽  
Praneetha Palasuberniam ◽  
Kae Yi Tan
Keyword(s):  
Snake Venom ◽  
Serine Proteases ◽  
Southern China ◽  
Snake Venoms ◽  
Phospholipases A2 ◽  
Limited Efficacy ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Species Specific ◽  
Venom Proteins

Snakebite envenomation is a serious neglected tropical disease, and its management is often complicated by the diversity of snake venoms. In Asia, pit vipers of the Ovophis species complex are medically important venomous snakes whose venom properties have not been investigated in depth. This study characterized the venom proteomes of Ovophis convictus (West Malaysia), Ovophis tonkinensis (northern Vietnam, southern China), and Ovophis okinavensis (Okinawa, Japan) by applying liquid chromatography-tandem mass spectrometry, which detected a high abundance of snake venom serine proteases (SVSP, constituting 40–60% of total venom proteins), followed by phospholipases A2, snake venom metalloproteinases of mainly P-III class, L-amino acid oxidases, and toxins from other protein families which were less abundant. The venoms exhibited different procoagulant activities in human plasma, with potency decreasing from O. tonkinensis > O. okinavensis > O. convictus. The procoagulant nature of venom confirms that consumptive coagulopathy underlies the pathophysiology of Ovophis pit viper envenomation. The hetero-specific antivenoms Gloydius brevicaudus monovalent antivenom (GbMAV) and Trimeresurus albolabris monovalent antivenom (TaMAV) were immunoreactive toward the venoms, and cross-neutralized their procoagulant activities, albeit at variably limited efficacy. In the absence of species-specific antivenom, these hetero-specific antivenoms may be useful in treating coagulotoxic envenomation caused by the different snakes in their respective regions.

scholarly journals Protein Identification of Venoms of the African Spitting Cobras, Naja mossambica and Naja nigricincta nigricincta

Toxins ◽  
2020 ◽  
Vol 12 (8) ◽  
pp. 520
Author(s):  
Ottilie Katali ◽  
Loide Shipingana ◽  
Peter Nyarangó ◽  
Mirva Pääkkönen ◽  
Erastus Haindongo ◽  
...  
Keyword(s):  
High Performance ◽  
Protein Identification ◽  
Sodium Dodecyl ◽  
Secretory Proteins ◽  
Tandem Mass ◽  
Protein Families ◽  
Chromatography Tandem Mass Spectrometry ◽  
Liquid Chromatography Tandem Mass ◽  
Fundamental Value ◽  
Venom Proteins

Cobra snakes, including Naja mossambica and Naja nigricincta nigricincta, are one of the major groups of snakes responsible for snakebites in southern Africa, producing significant cytotoxicity and tissue damage. The venom of N. mossambica has been briefly characterised, but that of N. n. nigricincta is not reported. The current study identifies the venom proteins of N. mossambica and N. n. nigricincta. This is achieved using sodium dodecyl sulphate (SDS)-polyacrylamide gel eletrophroresis (PAGE), followed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Most of the proteins were less than 17 kDa in both snakes. N. mossambica was found to have 75 proteins in total (from 16 protein families), whereas N.n. nigricincta had 73 (from 16 protein families). Of these identified proteins, 57 were common in both snakes. The proteins identified belonged to various families, including the three-finger toxins (3FTx), Cysteine-rich secretory proteins (CRiSP), Phospholipase A2 (PLA2) and Venom metalloproteinase M12B (SVMP). The current study contributes to the profile knowledge of snake venom compositions, which is of fundamental value in understanding the proteins that play a major role in envenomation.

Role of heme modulation in inhibition of Atheris, Atractaspis, Causus, Cerastes, Echis, and Macrovipera hemotoxic venom activity

2018 ◽  
Vol 38 (2) ◽  
pp. 216-226 ◽  
Author(s):  
VG Nielsen ◽  
N Frank
Keyword(s):  
Potential Therapeutic Target ◽  
Venomous Snake ◽  
Human Thrombin ◽  
Specific Variation ◽  
Coagulation Kinetics ◽  
Species Specific ◽  
Kinetics Of

Venomous snake bite and subsequent coagulopathy is a significant source of morbidity and mortality worldwide. The gold standard to treat coagulopathy caused by these venoms is the administration of antivenom; however, despite this therapy, coagulopathy still occurs and recurs. Of interest, our laboratory has demonstrated in vitro and in vivo that coagulopathy-inducing venom exposed to carbon monoxide (CO) is inhibited, potentially by an attached heme. The present investigation sought to determine if venoms derived from snakes of the African genera Atheris, Atractaspis, Causus, Cerastes, Echis, and Macrovipera that have no or limited antivenoms available could be inhibited with CO or with the metheme-inducing agent, O-phenylhydroxylamine (PHA). Assessing changes in coagulation kinetics of human plasma with thrombelastography, venoms were exposed in isolation to CO or PHA. Eight species were found to have procoagulant activity consistent with the generation of human thrombin, while one was likely fibrinogenolytic. All venoms were significantly inhibited by CO/PHA with species-specific variation noted. These data demonstrate indirectly that the heme is likely bound to these disparate venoms as an intermediary modulatory molecule. In conclusion, future investigation is warranted to determine if heme could serve as a potential therapeutic target to be modulated during treatment of envenomation by hemotoxic enzymes.

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