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Stephen P Mackessy - One of the best experts on this subject based on the ideXlab platform.
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Venom ontogeny in the mexican lance headed rattlesnake crotalus polystictus
Toxins, 2018Co-Authors: Stephen P Mackessy, Jamie Leroy, Estrella Mocinodeloya, Kirk Setser, Robert W Bryson, Anthony J SaviolaAbstract:As trophic adaptations, rattlesnake Venoms can vary in composition depending on several intrinsic and extrinsic factors. Ontogenetic changes in Venom composition have been documented for numerous species, but little is known of the potential age-related changes in many rattlesnake species found in Mexico. In the current study, Venom samples collected from adult and neonate Crotalus polystictus from Estado de Mexico were subjected to enzymatic and electrophoretic analyses, toxicity assays (LD50), and MALDI-TOF mass spectrometry, and a pooled sample of adult Venom was analyzed by shotgun proteomics. Electrophoretic profiles of adult males and females were quite similar, and only minor sex-based variation was noted. However, distinct differences were observed between Venoms from adult females and their neonate offspring. Several prominent bands, including P-I and P-III snake Venom metalloproteinases (SVMPs) and disintegrins (confirmed by MS/MS) were present in adult Venoms and absent/greatly reduced in neonate Venoms. Age-dependent differences in SVMP, kallikrein-like, phospholipase A2 (PLA2), and L-amino acid oxidase (LAAO) activity levels were confirmed by enzymatic activity assays, and like many other rattlesnake species, Venoms from adult snakes have higher SVMP activity than neonate Venoms. Conversely, PLA2 activity was approximately 2.5 × greater in Venoms from neonates, likely contributing to the increased toxicity (neonate Venom LD50 = 4.5 μg/g) towards non-Swiss albino mice when compared to adult Venoms (LD50 = 5.5 μg/g). Thrombin-like (TLE) and phosphodiesterase activities did not vary significantly with age. A significant effect of sex (between adult male and adult female Venoms) was also observed for SVMP, TLE, and LAAO activities. Analysis of pooled adult Venom by LC-MS/MS identified 14 toxin protein families, dominated by bradykinin-inhibitory peptides, SVMPs (P-I, P-II and P-III), disintegrins, PLA2s, C-type-lectins, CRiSPs, serine proteinases, and LAAOs (96% of total Venom proteins). Neonate and adult C. polystictus in this population consume almost exclusively mammals, suggesting that age-based differences in composition are related to physical differences in prey (e.g., surface-to-volume ratio differences) rather than taxonomic differences between prey. Venoms from adult C. polystictus fit a Type I pattern (high SVMP activity, lower toxicity), which is characteristic of many larger-bodied rattlesnakes of North America.
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Venom Ontogeny in the Mexican Lance-Headed Rattlesnake (Crotalus polystictus)
MDPI AG, 2018Co-Authors: Stephen P Mackessy, Jamie Leroy, Kirk Setser, Robert W Bryson, Estrella Mociño-deloya, Anthony J SaviolaAbstract:As trophic adaptations, rattlesnake Venoms can vary in composition depending on several intrinsic and extrinsic factors. Ontogenetic changes in Venom composition have been documented for numerous species, but little is known of the potential age-related changes in many rattlesnake species found in México. In the current study, Venom samples collected from adult and neonate Crotalus polystictus from Estado de México were subjected to enzymatic and electrophoretic analyses, toxicity assays (LD50), and MALDI-TOF mass spectrometry, and a pooled sample of adult Venom was analyzed by shotgun proteomics. Electrophoretic profiles of adult males and females were quite similar, and only minor sex-based variation was noted. However, distinct differences were observed between Venoms from adult females and their neonate offspring. Several prominent bands, including P-I and P-III snake Venom metalloproteinases (SVMPs) and disintegrins (confirmed by MS/MS) were present in adult Venoms and absent/greatly reduced in neonate Venoms. Age-dependent differences in SVMP, kallikrein-like, phospholipase A2 (PLA2), and L-amino acid oxidase (LAAO) activity levels were confirmed by enzymatic activity assays, and like many other rattlesnake species, Venoms from adult snakes have higher SVMP activity than neonate Venoms. Conversely, PLA2 activity was approximately 2.5 × greater in Venoms from neonates, likely contributing to the increased toxicity (neonate Venom LD50 = 4.5 μg/g) towards non-Swiss albino mice when compared to adult Venoms (LD50 = 5.5 μg/g). Thrombin-like (TLE) and phosphodiesterase activities did not vary significantly with age. A significant effect of sex (between adult male and adult female Venoms) was also observed for SVMP, TLE, and LAAO activities. Analysis of pooled adult Venom by LC-MS/MS identified 14 toxin protein families, dominated by bradykinin-inhibitory peptides, SVMPs (P-I, P-II and P-III), disintegrins, PLA2s, C-type-lectins, CRiSPs, serine proteinases, and LAAOs (96% of total Venom proteins). Neonate and adult C. polystictus in this population consume almost exclusively mammals, suggesting that age-based differences in composition are related to physical differences in prey (e.g., surface-to-volume ratio differences) rather than taxonomic differences between prey. Venoms from adult C. polystictus fit a Type I pattern (high SVMP activity, lower toxicity), which is characteristic of many larger-bodied rattlesnakes of North America
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colubrid Venom composition an omics perspective
Toxins, 2016Co-Authors: Inacio L M Junqueiradeazevedo, Pollyanna Fernandes Campos, Ana Tung Ching Ching, Stephen P MackessyAbstract:Snake Venoms have been subjected to increasingly sensitive analyses for well over 100 years, but most research has been restricted to front-fanged snakes, which actually represent a relatively small proportion of extant species of advanced snakes. Because rear-fanged snakes are a diverse and distinct radiation of the advanced snakes, understanding Venom composition among “colubrids” is critical to understanding the evolution of Venom among snakes. Here we review the state of knowledge concerning rear-fanged snake Venom composition, emphasizing those toxins for which protein or transcript sequences are available. We have also added new transcriptome-based data on Venoms of three species of rear-fanged snakes. Based on this compilation, it is apparent that several components, including cysteine-rich secretory proteins (CRiSPs), C-type lectins (CTLs), CTLs-like proteins and snake Venom metalloproteinases (SVMPs), are broadly distributed among “colubrid” Venoms, while others, notably three-finger toxins (3FTxs), appear nearly restricted to the Colubridae (sensu stricto). Some putative new toxins, such as snake Venom matrix metalloproteinases, are in fact present in several colubrid Venoms, while others are only transcribed, at lower levels. This work provides insights into the evolution of these toxin classes, but because only a small number of species have been explored, generalizations are still rather limited. It is likely that new Venom protein families await discovery, particularly among those species with highly specialized diets.
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Full-Length Venom Protein cDNA Sequences from Venom-Derived mRNA: Exploring Compositional Variation and Adaptive Multigene Evolution
2016Co-Authors: Cassandra M. Modahl, Stephen P MackessyAbstract:EnVenomation of humans by snakes is a complex and continuously evolving medical emergency, and treatment is made that much more difficult by the diverse biochemical composition of many Venoms. Venomous snakes and their Venoms also provide models for the study of molecular evolutionary processes leading to adaptation and genotype-phenotype relationships. To compare Venom complexity and protein sequences, Venom gland transcriptomes are assembled, which usually requires the sacrifice of snakes for tissue. However, toxin transcripts are also present in Venoms, offering the possibility of obtaining cDNA sequences directly from Venom. This study provides evidence that unknown full-length Venom protein transcripts can be obtained from the Venoms of multiple species from all major Venomous snake families. These unknown Venom protein cDNAs are obtained by the use of primers designed from conserved signal peptide sequences within each Venom protein superfamily. This technique was used to assemble a partial Venom gland transcriptome for the Middle American Rattlesnake (Crotalus simus tzabcan) by amplifying sequences for phospholipases A2, serine proteases, C-lectins, and metalloproteinases from within Venom. Phospholipase A2 sequences were also recovered from the Venoms of several rattlesnakes and an elapid snake (Pseudechis porphyriacus), and three-finger toxin sequences were recovered from multiple rear-fanged snake species, demonstrating that the three major clades of advanced snakes (Elapidae, Viperidae, Colubridae) have stable mRNA present in their Venoms. These cDNA sequences from Venom were then used to explore potential activities derived from protein sequence similarities and evolutionary histories within these large multigene superfamilies. Venom-derived sequences can also be used to aid in characterizing Venoms that lack proteomic profiles and identify sequence characteristics indicating specific enVenomation profiles. This approach, requiring only Venom, provides access to cDNA sequences in the absence of living specimens, even from commercial Venom sources, to evaluate important regional differences in Venom composition and to study snake Venom protein evolution.
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evolutionary trends in Venom composition in the western rattlesnakes crotalus viridis sensu lato toxicity vs tenderizers
Toxicon, 2010Co-Authors: Stephen P MackessyAbstract:The Western Rattlesnake (Crotalus viridis sensu lato, now including Crotalus oreganus )i s broadly distributed across the western half of the United States, northwestern Mexico and southwestern Canada, and eight subspecies are currently recognized. Although some Venom characteristics have been noted for most subspecies, a systematic study of Venoms from all subspecies has not been reported. Venom was extracted from snakes collected from approximate geographic range centers for all subspecies and analyzed using SDSPAGE, MALDI-TOF mass spectrometry, enzyme and toxicity assays. Electrophoretic and mass spectrometric analyses demonstrated that small myotoxins, disintegrins and PLA2 were abundant in most Venoms. PIII and PI metalloproteinases (w54 kDa and 23 kDa, respectively) were common to all Venoms except C. o. concolor, C. o. caliginis and C.o. helleri. Metalloproteinase activity was highest in C. o. cerberus and lowest in C. o. concolor Venoms (w100-fold difference). Conversely, C. o. concolor Venom was the most toxic and C. o. cerberus Venom was least toxic (15-fold difference). In general, Venoms with high metalloproteinase activity were less toxic (type I Venoms), while Venoms which were highly toxic showed low protease activity (type II Venoms). Within the C. viridis/oreganus complex, these two extremes of Venom compositional phenotypes are observed, and it appears that high metalloproteinase activity and high toxicity are incompatible qualities of these Venoms. The functional significance of these biochemical characteristics likely relates to characteristics of prey consumed, and Venoms with low metalloproteinase activity may constrain snake prey selection or foraging activity patterns.
Juan J. Calvete - One of the best experts on this subject based on the ideXlab platform.
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Venomic Analysis of the Poorly Studied Desert Coral Snake, Micrurus tschudii tschudii, Supports the 3FTx/PLA2 Dichotomy across Micrurus Venoms
Toxins, 2016Co-Authors: Libia Sanz, Alicia Perez, Davinia Pla, Bruno Lomonte, María Salas, Alfonso Zavaleta, Yania Rodriguez, Juan J. CalveteAbstract:The Venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a Venomic approach, which identified ≥38 proteins belonging to only four snake Venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the Venom proteome), the major protein family of the desert coral snake Venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the Venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent Venom phenotypes, i.e., 3FTx- and PLA2-predominant Venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of Venom phenotypic variability among true sea snake (Hydrophiinae) Venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae Venoms.
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Venomic analysis of the poorly studied desert coral snake micrurus tschudii tschudii supports the 3ftx pla2 dichotomy across micrurus Venoms
Toxins, 2016Co-Authors: Libia Sanz, Alicia Perez, Davinia Pla, Bruno Lomonte, María Salas, Alfonso Zavaleta, Yania Rodriguez, Juan J. CalveteAbstract:The Venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a Venomic approach, which identified ≥38 proteins belonging to only four snake Venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the Venom proteome), the major protein family of the desert coral snake Venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the Venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent Venom phenotypes, i.e., 3FTx- and PLA2-predominant Venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of Venom phenotypic variability among true sea snake (Hydrophiinae) Venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae Venoms.
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Snake Venomics of Micrurus alleni and Micrurus mosquitensis from the Caribbean region of Costa Rica reveals two divergent compositional patterns in New World elapids.
Toxicon : official journal of the International Society on Toxinology, 2015Co-Authors: Julián Fernández, José María Gutiérrez, Nancy Vargas-vargas, Davinia Pla, Mahmood Sasa, Paola Rey-suárez, Libia Sanz, Juan J. Calvete, Bruno LomonteAbstract:Protein composition, toxicity, and neutralization of the Venoms of Micrurus alleni and Micrurus mosquitensis, two sympatric monadal coral snakes found in humid environments of the Caribbean region of Costa Rica, were studied. Proteomic profiling revealed that these Venoms display highly divergent compositions: the former dominated by three-finger toxins (3FTx) and the latter by phospholipases A2 (PLA2). Protein family abundances correlated with enzymatic and toxic characteristics of the Venoms. Selective inhibition experiments showed that PLA2s play only a marginal role in the lethal effect of M. alleni Venom, but have a major role in M. mosquitensis Venom. Proteomic data gathered from other Micrurus species evidenced that the two divergent Venom phenotypes are recurrent, and may constitute a general trend across New World elapids. Further, M. mosquitensis, but not M. alleni, Venom contains PLA2-like/Kunitz-type inhibitor complex(es) that resemble the ASIC1a/2-activating MitTx heterodimeric toxin isolated from Micrurus tener Venom. The evolutionary origin and adaptive relevance of the puzzling phenotypic variability of Micrurus Venoms remain to be understood. An antiVenom against the PLA2-predominant Micrurus nigrocinctus Venom strongly cross-recognized and neutralized M. mosquitensis Venom, but only weakly M. alleni Venom.
Alicia Perez - One of the best experts on this subject based on the ideXlab platform.
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Venomic Analysis of the Poorly Studied Desert Coral Snake, Micrurus tschudii tschudii, Supports the 3FTx/PLA2 Dichotomy across Micrurus Venoms
Toxins, 2016Co-Authors: Libia Sanz, Alicia Perez, Davinia Pla, Bruno Lomonte, María Salas, Alfonso Zavaleta, Yania Rodriguez, Juan J. CalveteAbstract:The Venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a Venomic approach, which identified ≥38 proteins belonging to only four snake Venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the Venom proteome), the major protein family of the desert coral snake Venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the Venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent Venom phenotypes, i.e., 3FTx- and PLA2-predominant Venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of Venom phenotypic variability among true sea snake (Hydrophiinae) Venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae Venoms.
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Venomic analysis of the poorly studied desert coral snake micrurus tschudii tschudii supports the 3ftx pla2 dichotomy across micrurus Venoms
Toxins, 2016Co-Authors: Libia Sanz, Alicia Perez, Davinia Pla, Bruno Lomonte, María Salas, Alfonso Zavaleta, Yania Rodriguez, Juan J. CalveteAbstract:The Venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a Venomic approach, which identified ≥38 proteins belonging to only four snake Venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the Venom proteome), the major protein family of the desert coral snake Venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the Venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent Venom phenotypes, i.e., 3FTx- and PLA2-predominant Venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of Venom phenotypic variability among true sea snake (Hydrophiinae) Venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae Venoms.
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snake Venomics and antiVenomics of crotalus durissus subspecies from brazil assessment of geographic variation and its implication on snakebite management
Journal of Proteomics, 2010Co-Authors: Johara Boldrinifranca, Carlos Correanetto, Marliete Silva, Renata Santos Rodrigues, Pilar De La Torre, Alicia Perez, Andreimar M Soares, Russolina B Zingali, Romildo De Albuquerque Nogueira, Veridiana M RodriguesAbstract:Abstract We report the comparative proteomic and antiVenomic characterization of the Venoms of subspecies cascavella and collilineatus of the Brazilian tropical rattlesnake Crotalus durissus . The Venom proteomes of C. d. collilineatus and C. d. cascavella comprise proteins in the range of 4–115 kDa belonging to 9 and 8 toxin families, respectively. Collilineatus and cascavella Venoms contain 20–25 main toxins belonging to the following protein families: disintegrin, PLA 2 , serine proteinase, cysteine-rich secretory protein (CRISP), vascular endothelial growth factor-like (VEGF), l -amino acid oxidase, C-type lectin-like, and snake Venom metalloproteinase (SVMP). As judged by reverse-phase HPLC and mass spectrometry, cascavella and collilineatus share about 90% of their Venom proteome. However, the relative occurrence of the toxin families departs among the two C. durissus subspecies Venoms. The most notable difference is the presence of the myotoxin crotamine in some C. d. collilineatus specimens (averaging 20.8% of the total proteins of pooled Venom), which is absent in the Venom of C. d. cascavella. On the other hand, the neurotoxic PLA 2 crotoxin represents the most abundant protein in both C. durissus Venoms, comprising 67.4% of the toxin proteome in C. d. collilineatus and 72.5% in C. d. cascavella . Myotoxic PLA 2 s are also present in the two Venoms albeit in different relative concentrations (18.1% in C. d. cascavella vs. 4.6% in C. d. collilineatus ). The Venom composition accounts for the clinical manifestations caused by C. durissus enVenomations: systemic neurotoxicity and myalgic symptoms and coagulation disturbances, frequently accompanied by myoglobinuria and acute renal failure. The overall compositions of C. d. subspecies cascavella and collilineatus Venoms closely resemble that of C. d. terrificus , supporting the view that these taxa can be considered geographical variations of the same species. Pooled Venom from adult C.d. cascavella and neonate C.d. terrificus lack crotamine, whereas this skeletal muscle cell membrane depolarizing inducing myotoxin accounts for ∼ 20% of the total toxins of Venom pooled from C.d. collilineatus and C.d. terrificus from Southern Brazil. The possible relevance of the observed Venom variability among the tropical rattlesnake subspecies was assessed by antiVenomics using anti-crotalic antiVenoms produced at Instituto Butantan and Instituto Vital Brazil. The results revealed that both antiVenoms exhibit impaired immunoreactivity towards crotamine and display restricted (∼ 60%) recognition of PLA 2 molecules (crotoxin and D49-myotoxins) from C. d. cascavella and C. d. terrificus Venoms. This poor reactivity of the antiVenoms may be due to a combination of factors: on the one hand, an inappropriate choice of the mixture of Venoms for immunization and, on the other hand, the documented low immunogenicity of PLA 2 molecules. C. durissus causes most of the lethal snakebite accidents in Brazil. The implication of the geographic variation of Venom composition for the treatment of bites by different C. durissus subspecies populations is discussed.
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snake Venomics and antiVenomics of crotalus durissus subspecies from brazil assessment of geographic variation and its implication on snakebite management
Journal of Proteomics, 2010Co-Authors: Johara Boldrinifranca, Carlos Correanetto, Marliete Silva, Renata Santos Rodrigues, Pilar De La Torre, Alicia Perez, Andreimar M Soares, Russolina B Zingali, Romildo De Albuquerque Nogueira, Veridiana M RodriguesAbstract:We report the comparative proteomic and antiVenomic characterization of the Venoms of subspecies cascavella and collilineatus of the Brazilian tropical rattlesnake Crotalus durissus. The Venom proteomes of C. d. collilineatus and C. d. cascavella comprise proteins in the range of 4-115 kDa belonging to 9 and 8 toxin families, respectively. Collilineatus and cascavella Venoms contain 20-25 main toxins belonging to the following protein families: disintegrin, PLA(2), serine proteinase, cysteine-rich secretory protein (CRISP), vascular endothelial growth factor-like (VEGF), L-amino acid oxidase, C-type lectin-like, and snake Venom metalloproteinase (SVMP). As judged by reverse-phase HPLC and mass spectrometry, cascavella and collilineatus share about 90% of their Venom proteome. However, the relative occurrence of the toxin families departs among the two C. durissus subspecies Venoms. The most notable difference is the presence of the myotoxin crotamine in some C. d. collilineatus specimens (averaging 20.8% of the total proteins of pooled Venom), which is absent in the Venom of C. d. cascavella. On the other hand, the neurotoxic PLA(2) crotoxin represents the most abundant protein in both C. durissus Venoms, comprising 67.4% of the toxin proteome in C. d. collilineatus and 72.5% in C. d. cascavella. Myotoxic PLA(2)s are also present in the two Venoms albeit in different relative concentrations (18.1% in C. d. cascavella vs. 4.6% in C. d. collilineatus). The Venom composition accounts for the clinical manifestations caused by C. durissus enVenomations: systemic neurotoxicity and myalgic symptoms and coagulation disturbances, frequently accompanied by myoglobinuria and acute renal failure. The overall compositions of C. d. subspecies cascavella and collilineatus Venoms closely resemble that of C. d. terrificus, supporting the view that these taxa can be considered geographical variations of the same species. Pooled Venom from adult C.d. cascavella and neonate C.d. terrificus lack crotamine, whereas this skeletal muscle cell membrane depolarizing inducing myotoxin accounts for approximately 20% of the total toxins of Venom pooled from C.d. collilineatus and C.d. terrificus from Southern Brazil. The possible relevance of the observed Venom variability among the tropical rattlesnake subspecies was assessed by antiVenomics using anti-crotalic antiVenoms produced at Instituto Butantan and Instituto Vital Brazil. The results revealed that both antiVenoms exhibit impaired immunoreactivity towards crotamine and display restricted ( approximately 60%) recognition of PLA(2) molecules (crotoxin and D49-myotoxins) from C. d. cascavella and C. d. terrificus Venoms. This poor reactivity of the antiVenoms may be due to a combination of factors: on the one hand, an inappropriate choice of the mixture of Venoms for immunization and, on the other hand, the documented low immunogenicity of PLA(2) molecules. C. durissus causes most of the lethal snakebite accidents in Brazil. The implication of the geographic variation of Venom composition for the treatment of bites by different C. durissus subspecies populations is discussed.
Bruno Lomonte - One of the best experts on this subject based on the ideXlab platform.
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ontogenetic changes in the Venom of metlapilcoatlus nummifer the mexican jumping viper
Toxicon, 2020Co-Authors: Belem Garciaosorio, Bruno Lomonte, Melisa Benardvalle, Jorge Lopez De Leon, Luis Romandominguez, Nancy R Mejiadominguez, Felipe Larahernandez, Alejandro Alagon, Edgar NericastroAbstract:The viperid genus Metlapilcoatlus (previously Atropoides) is represented in Mexico by four species: M. olmec, M. mexicanus, M. occidus, and M. nummifer. To date, no studies on their Venoms have been reported. Here, we comparatively characterized the Venom from M. nummifer neonates (≤8 months of age), young adults (18 months) and adults (≥24 months). We performed biological and enzymatic activities, as well as electrophoretic and RP-HPLC profiling combined with proteomic assignment of major fractions. Venoms from neonates and adults differed in their electrophoretic and chromatographic profiles, indicating that an ontogenetic compositional shift occurs in this species. Protein family assignments showed that neonates produce a Venom rich in Snake Venom Metalloproteinases (SVMPs) and Snake Venom Serine Proteases (SVSPs), but lacking Phospholipases A2 (PLA2s). In contrast, adults express abundant Venom PLA2s, and lower molecular weight proteins, as evidenced by SDS-PAGE. Functionally, neonate Venom did not display PLA2 or procoagulant activities, whereas adult Venom did. Hemorrhagic activity was present in both neonate and adult Venoms, with similar potencies. Finally, it is of considerable concern that the lethal activity of neither neonate nor adult Venoms was neutralized by two therapeutic antiVenoms produced in Mexico.
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biological and proteolytic variation in the Venom of crotalus scutulatus scutulatus from mexico
Toxins, 2018Co-Authors: Miguel Borja, Bruno Lomonte, Edgar Nericastro, Gamaliel Castanedagaytan, Jason L Strickland, Christopher L Parkinson, Juan Castanedagaytan, Roberto Poncelopez, Alejandro Olverarodriguez, Alejandro AlagonAbstract:Rattlesnake Venoms may be classified according to the presence/absence and relative abundance of the neurotoxic phospholipases A 2 s (PLA 2 s), such as Mojave toxin, and snake Venom metalloproteinases (SVMPs). In Mexico, studies to determine Venom variation in Mojave Rattlesnakes (Crotalus scutulatus scutulatus) are limited and little is known about the biological and proteolytic activities in this species. Tissue (34) and Venom (29) samples were obtained from C. s. scutulatus from different locations within their distribution in Mexico. Mojave toxin detection was carried out at the genomic (by PCR) and protein (by ELISA) levels for all tissue and Venom samples. Biological activity was tested on representative Venoms by measuring LD 50 and hemorrhagic activity. To determine the approximate amount of SVMPs, 15 Venoms were separated by RP-HPLC and variation in protein profile and proteolytic activity was evaluated by SDS-PAGE (n = 28) and Hide Powder Azure proteolytic analysis (n = 27). Three types of Venom were identified in Mexico which is comparable to the intraspecific Venom diversity observed in the Sonoran Desert of Arizona, USA: Venom Type A (∼Type II), with Mojave toxin, highly toxic, lacking hemorrhagic activity, and with scarce proteolytic activity; Type B (∼Type I), without Mojave toxin, less toxic than Type A, highly hemorrhagic and proteolytic; and Type A + B, containing Mojave toxin, as toxic as Venom Type A, variable in hemorrhagic activity and with intermediate proteolytic activity. We also detected a positive correlation between SVMP abundance and hemorrhagic and proteolytic activities. Although more sampling is necessary, our results suggest that Venoms containing Mojave toxin and Venom lacking this toxin are distributed in the northwest and southeast portions of the distribution in Mexico, respectively, while an intergradation in the middle of both zones is present.
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Venomic Analysis of the Poorly Studied Desert Coral Snake, Micrurus tschudii tschudii, Supports the 3FTx/PLA2 Dichotomy across Micrurus Venoms
Toxins, 2016Co-Authors: Libia Sanz, Alicia Perez, Davinia Pla, Bruno Lomonte, María Salas, Alfonso Zavaleta, Yania Rodriguez, Juan J. CalveteAbstract:The Venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a Venomic approach, which identified ≥38 proteins belonging to only four snake Venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the Venom proteome), the major protein family of the desert coral snake Venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the Venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent Venom phenotypes, i.e., 3FTx- and PLA2-predominant Venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of Venom phenotypic variability among true sea snake (Hydrophiinae) Venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae Venoms.
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Venomic analysis of the poorly studied desert coral snake micrurus tschudii tschudii supports the 3ftx pla2 dichotomy across micrurus Venoms
Toxins, 2016Co-Authors: Libia Sanz, Alicia Perez, Davinia Pla, Bruno Lomonte, María Salas, Alfonso Zavaleta, Yania Rodriguez, Juan J. CalveteAbstract:The Venom proteome of the poorly studied desert coral snake Micrurus tschudii tschudii was unveiled using a Venomic approach, which identified ≥38 proteins belonging to only four snake Venom protein families. The three-finger toxins (3FTxs) constitute, both in number of isoforms (~30) and total abundance (93.6% of the Venom proteome), the major protein family of the desert coral snake Venom. Phospholipases A2 (PLA2s; seven isoforms, 4.1% of the Venom proteome), 1–3 Kunitz-type proteins (1.6%), and 1–2 l-amino acid oxidases (LAO, 0.7%) complete the toxin arsenal of M. t. tschudii. Our results add to the growing evidence that the occurrence of two divergent Venom phenotypes, i.e., 3FTx- and PLA2-predominant Venom proteomes, may constitute a general trend across the cladogenesis of Micrurus. The occurrence of a similar pattern of Venom phenotypic variability among true sea snake (Hydrophiinae) Venoms suggests that the 3FTx/PLA2 dichotomy may be widely distributed among Elapidae Venoms.
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Snake Venomics of Micrurus alleni and Micrurus mosquitensis from the Caribbean region of Costa Rica reveals two divergent compositional patterns in New World elapids.
Toxicon : official journal of the International Society on Toxinology, 2015Co-Authors: Julián Fernández, José María Gutiérrez, Nancy Vargas-vargas, Davinia Pla, Mahmood Sasa, Paola Rey-suárez, Libia Sanz, Juan J. Calvete, Bruno LomonteAbstract:Protein composition, toxicity, and neutralization of the Venoms of Micrurus alleni and Micrurus mosquitensis, two sympatric monadal coral snakes found in humid environments of the Caribbean region of Costa Rica, were studied. Proteomic profiling revealed that these Venoms display highly divergent compositions: the former dominated by three-finger toxins (3FTx) and the latter by phospholipases A2 (PLA2). Protein family abundances correlated with enzymatic and toxic characteristics of the Venoms. Selective inhibition experiments showed that PLA2s play only a marginal role in the lethal effect of M. alleni Venom, but have a major role in M. mosquitensis Venom. Proteomic data gathered from other Micrurus species evidenced that the two divergent Venom phenotypes are recurrent, and may constitute a general trend across New World elapids. Further, M. mosquitensis, but not M. alleni, Venom contains PLA2-like/Kunitz-type inhibitor complex(es) that resemble the ASIC1a/2-activating MitTx heterodimeric toxin isolated from Micrurus tener Venom. The evolutionary origin and adaptive relevance of the puzzling phenotypic variability of Micrurus Venoms remain to be understood. An antiVenom against the PLA2-predominant Micrurus nigrocinctus Venom strongly cross-recognized and neutralized M. mosquitensis Venom, but only weakly M. alleni Venom.
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a survey on some biochemical and pharmacological activities of Venom from two colombian colubrid snakes erythrolamprus bizona double banded coral snake mimic and pseudoboa neuwiedii neuwied s false boa
Toxicon, 2017Co-Authors: Kristian A Torresbonilla, Rafael Stuani Floriano, Raphael Schezaroramos, Lea Rodriguessimioni, Maria Alice Da CruzhoflingAbstract:Abstract Colombian colubrid snake Venoms have been poorly studied. They represent a great resource of biological, ecological, toxinological and pharmacological research. We assessed some enzymatic properties and neuromuscular effects of Erythrolamprus bizona and Pseudoboa neuwiedii Venoms from Colombia. Proteolytic, amidolytic and phospholipase A 2 (PLA 2 ) activities were analyzed using colorimetric assays and the neuromuscular activity was analyzed in chick biventer cervicis (BC) preparations. The Venom of both species showed very low PLA 2 and amidolytic activities; however, both exhibited high proteolytic activity, which in E. bizona Venom surpassed that of P. neuwiedii Venom. E. bizona and P. neuwiedii Venoms provoked partial neuromuscular blockade, which was more prominent in P. neuwiedii Venom. E. bizona Venom (30 μg/ml) induced a significant potentiation of the contracture response to exogenous ACh (110 μM), which was not accompanied by twitch height alteration, whereas the highest Venom concentration (100 μg/ml) inhibited contracture responses to both ACh and KCl (40 mM). In contrast, P. neuwiedii Venom (30 and 100 μg/ml) caused significant reduction in the contracture responses to exogenous ACh and KCl. The morphological analyses showed high myotoxic effects in the muscle fibers of BC incubated with either Venoms; however, they are more prominent in the P. neuwiedii Venom. Our results suggest that the myotoxicity of the Venom of the two Colombian species can be ascribed to their high proteolytic activity. An interesting data was the potentiation of the ACh-induced contracture, but not the twitch height, caused by E. bizona Venom, at a concentration that is harmless to muscle fibers integrity. This phenomenon remains to be further elucidated, and suggest that a possible involvement of post-synaptic receptors cannot be discarded. This work is a contribution to expand the knowledge on colubrid Venoms; it allows envisaging that the two Venoms offer the potential to go further in the identification of their components and biological targets.
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a survey on some biochemical and pharmacological activities of duvernoy s gland secretions from two colombian colubrid snakes erythrolamprus bizona double banded coral snake mimic and pseudoboa neuwiedii neuwied s false boa
Toxicon, 2017Co-Authors: Kristian A Torresbonilla, Rafael Stuani Floriano, Raphael Schezaroramos, Lea Rodriguessimioni, Maria Alice Da CruzhoflingAbstract:Colombian colubrid snake Venoms have been poorly studied. They represent a great resource of biological, ecological, toxinological and pharmacological research. We assessed some enzymatic properties and neuromuscular effects of Erythrolamprus bizona and Pseudoboa neuwiedii Venoms from Colombia. Proteolytic, amidolytic and phospholipase A2 (PLA2) activities were analyzed using colorimetric assays and the neuromuscular activity was analyzed in chick biventer cervicis (BC) preparations. The Venom of both species showed very low PLA2 and amidolytic activities; however, both exhibited high proteolytic activity, which in E. bizona Venom surpassed that of P. neuwiedii Venom. E. bizona and P. neuwiedii Venoms provoked partial neuromuscular blockade, which was more prominent in P. neuwiedii Venom. E. bizona Venom (30 μg/ml) induced a significant potentiation of the contracture response to exogenous ACh (110 μM), which was not accompanied by twitch height alteration, whereas the highest Venom concentration (100 μg/ml) inhibited contracture responses to both ACh and KCl (40 mM). In contrast, P. neuwiedii Venom (30 and 100 μg/ml) caused significant reduction in the contracture responses to exogenous ACh and KCl. The morphological analyses showed high myotoxic effects in the muscle fibers of BC incubated with either Venoms; however, they are more prominent in the P. neuwiedii Venom. Our results suggest that the myotoxicity of the Venom of the two Colombian species can be ascribed to their high proteolytic activity. An interesting data was the potentiation of the ACh-induced contracture, but not the twitch height, caused by E. bizona Venom, at a concentration that is harmless to muscle fibers integrity. This phenomenon remains to be further elucidated, and suggest that a possible involvement of post-synaptic receptors cannot be discarded. This work is a contribution to expand the knowledge on colubrid Venoms; it allows envisaging that the two Venoms offer the potential to go further in the identification of their components and biological targets.
