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At least 883 records · Page 49Linked to original sources
PubMed1995

The molecular cloning of a phospholipase A2 from Bothrops jararacussu snake venom: evolution of venom group II phospholipase A2's may imply gene duplications.

The sequence coding for a snake venom phospholipase A2 (PLA2), BJUPLA2, has been cloned from a Bothrops jararacussu venom gland cDNA library. The cDNA sequence predicts a precursor containing a 16-residue signal peptide followed by a molecule of 122 amino acid residues with a strong sequence similarity to group II snake venom PLA2's. A striking feature of the cDNA is the high sequence conservation of the 5' and 3' untranslated regions in cDNAs coding for PLA2's from a number of viper species. The greatest sequence variation was observed between the regions coding for the mature proteins, with most substitutions occurring in nonsynonymous sites. The phylogenetic tree constructed by alignment of the amino acid sequence of BJUPLA2 with group II PLA2's in general groups them according to current taxonomical divisions and/or functional activity. It also suggests that gene duplications may have occurred at a number of different points during the evolution of snake venom group II PLA2's.

Amino Acid Sequence↗
PubMed1996

Angler Survey Contributes to Socially Acceptable Modification of Harvest Regulations to Preserve Cutthroat Trout Fishery in Snake River, Wyoming, USA

This is a case study that describes a survey of anglers that was used to assist in modifying fishing regulations for indigenous trout in the Snake River, Wyoming. A mail survey of anglers who purchased 1991 Wyoming fishing licenses in the two counties adjacent to the Snake River was conducted during fall 1992. Differences in angler preferences were noted between anglers who purchased licenses in two adjacent counties with different socioeconomic structures, as well as between residents and nonresidents in each county. Anglers who purchased licenses in Teton County, where there is extensive tourism and immigration by relatively wealthy residents, tended to be more specialized and less harvest oriented. Anglers in Lincoln County, which is largely agricultural and has substantially less tourism and immigration of residents, tended to fish in many different ways and indicated more desire to harvest fish. Anglers from the two counties segregated themselves; those from Teton County primarily used the upstream portion of the study reach, and those from Lincoln County primarily used a short downstream portion of the reach. Modification of fishing regulations to reduce harvest of spawning-size cutthroat trout in the Snake River probably was acceptable to most anglers due to spatial segregation and their attitudes toward harvest.KEY WORDS: Rivers; Salmonidae; Trout; Anglers; Regulations; Wyoming

Journal Article↗
PubMed2001

Pulmonary receptors in reptiles: discharge patterns of receptor populations in snakes versus turtles.

This study examines the effects of lung inflation/deflation with and without CO2 on the entire population of pulmonary receptors in the vagus nerve in two species of snakes and two species of turtles. We asked the question, "how does the response of the entire mixed population of pulmonary stretch receptors (PSR) and intrapulmonary chemoreceptors (IPC) in species possessing both differ from that in species with only PSR"? This was studied under conditions of artificial ventilation with the secondary goal of extending observations on the presence/absence of IPC to a further three species. Our results indirectly illustrate the presence of IPC in the Burmese python and South American rattlesnake but not the side necked turtle, adding support to the hypothesis that IPC first arose in diapsid reptiles. In both species of snake, CO2-sensitive discharge (presumably from IPC) predominated almost to the exclusion of CO2-insensitive discharge (presumably arising from PSR) while the opposite was true for both species of turtle. The data suggest that for animals breathing air under conditions of normal metabolism there is little to distinguish between the discharge profiles of the total population of receptors arising from the lungs in the different groups. Interestingly, however, under conditions of elevated environmental CO2 most volume-related feedback from the lungs is abolished in the two species of snakes, while under conditions of elevated metabolic CO2, it is estimated that volume feedback from the lungs would be enhanced in these same species.

Animals↗
PubMed2004

Snake infrared receptors respond to dimethylsulfoxide in the blood stream.

1. We used extracellular recording of the infrared (IR)-sensitive trigeminal ganglion (TG) neurons (primary neurons) of a crotaline snake, Trimeresurus flavoviridis, which has very sensitive thermoreceptors, to examine changes in the IR response induced by dimethylsulfoxide (DMSO), in vivo. 2. The responses in the TG were recorded after each concentration of DMSO (1, 10, and 25%) was administered in the bloodstream. 3. At a constant temperature, DMSO dose-dependently potentiated the IR-triggered discharges of IR-sensitive TG neurons in this snake. 4. It is suggested that the increased IR response to DMSO is due to its chemical effect, or to an indirect effect via its vasoactive role in the thermoreceptors of IR-sensitive snakes.

Action Potentials↗
PubMed1984

Properties and applications of immobilized snake venom NAD glycohydrolase.

The NAD glycohydrolase (NADase) from Bungarus fasciatus snake venom was adsorbed on concanavalin A-Sepharose, and demonstrated to retain both hydrolase and transglycosidase activities in the bound form. The matrix-bound enzyme was stable to repeated washing with buffer and storage at 4 degrees C. The bound enzyme exhibited the same Km value for hydrolysis of nicotinamide-1,N6-ethenoadenine dinucleotide as previously measured with the soluble, purified form of the enzyme. The bound NADase was used repeatedly for a preparative-scale synthesis of 3-acetylpyridine adenine dinucleotide. It was further demonstrated that the immobilized enzyme could be prepared directly from crude snake venom, thus avoiding the time required for purification. The application of the immobilized snake venom NADase for the preparation of pyridine nucleotide coenzyme analogs has many advantages over procedures used previously for analog synthesis.

Catalysis↗
PubMed1995

Snake venom toxins, unlike smaller antagonists, appear to stabilize a resting state conformation of the nicotinic acetylcholine receptor.

Previous studies have shown that the pattern and degree of 3-(trifluoromethyl)-3-(m-[125I]iodophenyl)diazirine ([125I]TID) photoincorporation into the nicotinic acetylcholine receptor (nAChR) can be used as a sensitive measure of nAChR conformation. Upon desensitization by prolonged exposure to agonists, certain drugs and detergents, or reconstitution into desensitizing lipids, the levels of [125I]TID incorporation into the subunits of the nAChR are dramatically reduced. In this study, we characterized the effects of the snake venom proteins alpha-bungarotoxin and alpha-cobrotoxin, as well as the smaller antagonists tubocurarine and gallamine, on [125I]TID incorporation into the subunits of both partially-purified nAChR in native lipids, or affinity-purified nAChR reconstituted into different combinations of lipids. Unlike all other compounds previously tested, alpha-bungarotoxin and alpha-cobrotoxin reproducibly increased the level of [125I]TID incorporation into all four subunits of nAChR reconstituted into dioleoylphosphatidylcholine, dioleoylphosphatidic acid and cholesterol. Gallamine had little or no effect on [125I]TID incorporation at any concentration tested (0.1 microM-5 mM). Tubocurarine had no effect on [125I]TID incorporation at low concentrations, but at higher concentrations reduced the level of [125I]TID labeling. The snake venom proteins may shift the population of nAChR, which exists as a mixture of resting state and desensitized conformations, entirely to the resting state. However, the binding of the snake venom toxins does not appear sufficient to induce the resting state conformation in nAChR which have been desensitized by other means, such as solubilization in desensitizing detergents or reconstitution in densitizing lipids.

Affinity Labels↗
PubMed1993

Activation of snake venom metalloproteinases by a cysteine switch-like mechanism.

The cDNAs of several snake venom zinc endopeptidases code for a putative propeptide, which includes the conserved cysteine-containing sequence PKMCGVT. It has been suggested that binding of the cysteine thiol function to the active-site zinc, resulting in inactivation of the catalytic domain, occurs in a mode similar to the 'cysteine switch' mechanism proposed for matrix metalloproteinases. In order to confirm this hypothesis, inhibition kinetics have been performed on the metalloproteinase adamalysin II of the venom of the snake Crotalus adamanteus using several cysteine peptides. Among these the synthetic hexapeptide PKMCGV-NH2, corresponding to the conserved sequence portion of the known propeptides, was found to be by far the strongest inhibitor of this proteinase with a Ki of 3.4 microM. The inhibitory potencies of an equivalent peptide with the L-Cys replaced by a D-Cys or by an L-Ser as well as of reduced glutathione, cysteine and two unrelated cysteine peptides were by one to two orders of magnitudes lower. These findings strongly support a cysteine switch-like mechanism even for activation of the snake venom metalloproteinases.

Amino Acid Sequence↗
PubMed1990

Effect of snake venom of Agkistrodon halys on atherosclerosis and blood characteristics in Japanese quail.

Extracts of snake venom have been widely used for the treatment of vascular thrombotic diseases, yet the therapeutic mechanism is not clear. The effect of snake venom fractions on atherosclerosis in Japanese quail was studied. The venom of Agkistrodon halys was fractionated by DEAE-cellulose chromatography and the pooled protein fractions that resulted were injected intravenously into the quail with aortic atherosclerosis induced by dietary cholesterol. After 7 weeks of injections on every other day, the quail were killed, blood clotting times and serum cholesterol levels were determined, and aortic atherosclerosis and fatty liver were scored. The results showed that while no regression of atherosclerosis was observed, the lowering of serum cholesterol, prolonged blood clotting time and reduced fatty liver were significantly affected by the injection of one of the pooled protein fractions. This venom fraction contained two major protein components, one of which had arginine esterase activity. From this study we conclude that snake venom has little effect on the regression of atherosclerosis, but it prolongs blood clotting and lowers serum cholesterol.

Animals↗
PubMed1994

Purification and characterization of fibrolase isoforms from venom of individual southern copperhead (Agkistrodon contortrix Contortrix) snakes.

Fibrolase, a zinc metalloproteinase possessing direct-acting fibrinolytic activity, has been previously purified from southern copperhead (Agkistrodon contortrix contortix) snake venom. We recently reported that a pool of southern copperhead venom from different geographical locations possesses two isoforms of fibrolase (fib1 and fib2) [Loayza, S. L. et al. (1994) J. Chromat. B, in press]. We now report that venom from individual southern copperhead snakes contains the two isoforms which can be separated by a three-step high performance liquid chromatography (HPLC) procedure consisting of hydrophobic interaction chromatography, hydroxylapatite chromatography and weak cation exchange chromatography. Utilizing mass spectrometry we determined that fib1 has a molecular mass of 22,879 atomic mass units (amu) compared to 22,753 amu for fib2. These results support earlier observations during amino acid sequence analysis that a truncated version of the enzyme is produced which is missing the amino-terminal amino acid (< Glu-Arg-Phe-Pro vs. the intact enzyme < Glu-Gln-Arg-Phe-Pro, where < Glu is cyclized glutamine). The truncated version of fibrolase (fib2) has full fibrinolytic activity compared to fib1. EC50 values (concentration of enzyme required to degrade 50% of fibrin in a micro-fibrin plate assay) are 6.4 (+/- 1.0) microM and 5.2 (+/- 0.8) microM for fib 1 and fib2, respectively. Therefore, loss of the amino-terminal amino acid does not appear to influence enzymatic activity. We conclude that the two isoforms of fibrolase arise from variations in the molecular processing of the enzyme by the snake venom gland rather than being caused by the pooling of southern copperhead venoms from different geographical locations.

Agkistrodon↗
PubMed1994

Development of an enzyme-linked immunosorbent assay (ELISA) for identification of venoms from snakes in the Agkistrodon genus.

An enzyme-linked immunosorbent assay (ELISA) using a purified myotoxin from the venom of Agkistrodon contortrix laticintus (broad-banded copperhead) as immunogen was developed for potential use in the identification of envenomation by snakes belonging to the genus Agkistrodon native to North America. The specificity of the assay was tested using a total of 43 venom samples from snakes of diverse geographic locations. Venom samples used for cross-reactivity determination represent eight snake families including 14 species from the genus Crotalus. The assay detected venom from all Agkistrodon species tested without significant cross-reactivity with other venoms except for samples from two species of Bothrops which do not occur naturally north of Southern Mexico. The detection limit of the assay was 2 ng/ml for homologous crude venom dissolved in normal human serum. The assay was highly accurate in correlating optical densities with venom concentrations (r = 0.997). The presence of the antigen in experimental envenomations was readily detected by the assay at an i.m. injection dosage of 0.1 microgram/g. This ELISA is a promising test for identification of envenomations by species of Agkistrodon found in most of North America. It can also be used to study the kinetics of the myotoxin in experimental envenomations.

Agkistrodon↗
PubMed2005

Specific distribution of VEGF-F in Viperinae snake venoms: isolation and characterization of a VGEF-F from the venom of Daboia russelli siamensis.

Vascular endothelial growth factor exerts multiple functions through binding to two distinct receptor tyrosine kinases, Flt-1 and KDR. We previously demonstrated that snake venom VEGFs, designated VEGF-Fs, exhibit potent biological activities when compared with VEGF165 and that they selectively recognize KDR. We herein report the screening of several snake venoms for VEGF-Fs using antibodies against vammin and HF, VEGF-Fs derived from the venom of Vipera ammodytes ammodytes and Vipera aspis aspis, respectively. Specific immunoreactivity was observed in the venoms of Vipera and Daboia species. The present results suggest that VEGF-Fs are specific components of venoms from the Viperinae snakes that inhabit Europe, India, and Asia. We also report the isolation and characterization of an additional VEGF-F from Daboia russelli siamensis venom.

Amino Acid Sequence↗
PubMed2006

Characterization of a fibrinolytic enzyme (ussurenase) from Agkistrodon blomhoffii ussurensis snake venom: insights into the effects of Ca2+ on function and structure.

Fibrino(geno)lytic enzymes from snake venoms have been identified as high quality therapeutic agents for treatment of blood clots and strokes. They act on fibrinogen and fibrin, leading to defibrinogenation of blood, lysis of fibrin, and a consequent decrease in blood viscosity. In this work, a fibrinolytic enzyme (ussurenase) from China Agkistrodon blomhoffii Ussurensis snake venom, was purified to homogeneity, identified as a stable 23,367.8 Da monomeric protein, and was identified as a new kind of snake venom metalloproteinase. Ussurenase reacts optimally with fibrin clots at pH 7.5-8.3 and a temperature of 33-41 degrees C. Although many fibrinolytic enzymes are known to be zinc-dependent, measurements from inductively coupled plasma-atomic emission spectroscopy (ICP-AES) reveal that ussurenase is a Ca2+-containing protein with a molar ratio of 1:1 ([Ca2+]:[enzyme]). Ca2+ is crucial to the fibrin clot hydrolysis by ussurenase but also plays an important role in maintaining the structural integrity of the enzyme. The addition of Ca2+ to the apoenzyme induces a conformational change making the environments surrounding the Trp residues of the enzyme more hydrophobic. The presence of Ca2+ also increases the structural stability of ussurenase, so that higher concentrations of the denaturant guanidine hydrochloride are required to denature the native ussurenase compared to the apo-form. UV absorption and CD spectroscopy experiments show that Ca2+ increases the thermostability and changes the secondary structure of ussurenase. All these data suggest that Ca2+ is crucial for the correct folding and activity of ussurenase.

Agkistrodon↗
PubMed2004

S1 subsite in snake venom thrombin-like enzymes: can S1 subsite lipophilicity be used to sort binding affinities of trypsin-like enzymes to small-molecule inhibitors?

Thrombin-like enzymes isolated from snake venoms comprise a group of serine proteinases responsible for many important coagulation disorders in the envenomed victims. Besides, these proteinases have great biotechnological interest as antithrombotic agents and as diagnostic tools. However, in spite of the recent overflow of snake venom thrombin-like enzymes (SVTLEs) on protein sequence databases, there is a lack of three-dimensional (3D) structural information on this family. Without such 3D structures available many aspects of the biological function and biochemical properties of these enzymes still remain obscure. Therefore, we have gone through a series of computational techniques, which enabled us to identify the set of residues involved in molecular recognition of inhibitors bound to the S1 subsite of snake venom thrombin-like enzymes (SVTLEs) and ultimately conclude that nonpolar (van der Waals) intermolecular interactions and ligand's hydrophobicity are the most important factors affecting binding affinities to the S1 subsite of a SVTLE isolated from the venom of Lachesis muta muta (Lmm-TLE). Consequently, we have proposed that S1 subsite lipophilicity may be used to sort binding affinities of trypsin-like enzymes to small molecules by showing that the inhibitory potency of several S1-directed compounds follows subsite lipophilicity among Lmm-TLE and other three homologous proteases. Noteworthy, in the course of our analyses we determined that thrombin's S1 subsite should, in fact, be considered less lipophilic than that of trypsin if we account for the presence of the sodium-controlled water channel communicating with the S1 subsite in the coagulant enzyme.

Amidines↗
PubMed2004

Neo-clerodane diterpenoid, a new metalloprotease snake venom inhibitor from Baccharis trimera (Asteraceae): anti-proteolytic and anti-hemorrhagic properties.

Many plants are used in traditional medicine as active agents against various effects induced by snakebite. Few attempts have been made however to identify the nature of plant natural products with anti-ophidian properties. Baccharis trimera (Less) DC (Asteraceae), known in Brazil as carqueja, has been popularly used to treat liver diseases, rheumatism, diabetes, as well as digestive, hepatic and renal disorders. The active component was identified as 7alpha-hydroxy-3,13-clerodadiene-16,15:18,19-diolide, C20H28O5, (clerodane diterpenoid, Bt-CD). We report now the anti-proteolytic and anti-hemorrhagic properties against snake venoms of a Bt-CD inhibitor from B. trimera. Bt-CD exhibited full inhibition of hemorrhage and proteolytic activity caused by Bothrops snake venoms. The inhibitor was able to neutralize the hemorrhagic, fibrinogenolytic and caseinolytic activities of class P-I and III metalloproteases isolated from B. neuwiedi and B. jararacussu venoms. No inhibition of the coagulant activity was observed. Bt-CD also partially inhibited the edema induced by other crude venoms, metalloproteases, basic and acidic phospholipases A2. To further elucidate the inhibitory specificity of Bt-CD against metalloproteases isolated from snake venoms, a deeper understanding of its structure and function is necessary. Furthermore, the potential use of these inhibitors to complement anti-venom as an alternative treatment of snakebite envenomations needs to be evaluated in future studies.

Animals↗
PubMed2006

Isolation, characterization and molecular cloning of AnMIP, a new alpha-type phospholipase A2 myotoxin inhibitor from the plasma of the snake Atropoides nummifer (Viperidae: Crotalinae).

A new phospholipase A(2) (PLA(2))-inhibitory protein was isolated from the plasma of Atropoides nummifer, a crotaline snake from Central America. This inhibitor was named AnMIP, given its ability to neutralize the activity of basic PLA(2) myotoxins of its own and related venoms. The cDNA of AnMIP was cloned and sequenced, showing that it belongs to the alpha group of phospholipase A(2) inhibitors (PLIs). AnMIP appears as a homotrimer in the native state, held together by non-covalent forces, with a subunit molecular mass of 22,247-22,301 and an isoelectric point of 4.1-4.7. This trimeric structure is the first observed in a PLIalpha from American crotaline snakes, previously reported only in Asian species. Sequencing, mass spectrometry, and analytical isoelectrofocusing indicated the existence of isoforms, as reported for other PLIalphas isolated from snake plasma. The inhibitory profile of AnMIP showed specificity towards group II PLA(2)s, either belonging to the catalytically-active (D49) or -inactive (K49) subtypes, exemplified in this study by Bothrops asper myotoxin I and A. nummifer myotoxin II, respectively. By phylogenetic analysis it was shown that AnMIP is closely related to CgMIP-II, previously isolated from the plasma of Cerrophidion godmani, showing 93% amino acid sequence identity.

Amino Acid Sequence↗
PubMed2004

Effect of crotapotin on the biological activity of Asp49 and Lys49 phospholipases A(2) from Bothrops snake venoms.

Myonecrosis, in addition to edema and other biological manifestations, are conspicuous effects of Bothrops snake venoms, some of them caused by phospholipases A(2) (PLA(2)s). Asp49-PLA(2)s are catalytically active, whereas Lys49-PLA(2)s, although highly toxic, have little or no enzymatic activity upon artificial substrates, due to a substitution of lysine for aspartic acid at position 49. Crotapotin (CA), the acidic counterpart of crotoxin PLA(2) (CB), is a PLA(2)-like protein from Crotalus durissus terrificus snake venom, and is considered a chaperone protein for CB, able to increase its lethality about ten fold, but to inhibit the formation of the rat paw edema induced by carrageenin and by snake venoms. In this study, we demonstrate that CA significantly inhibits the edema induced by BthTX-I (23% inhibition), BthTX-II (27%), PrTX-I (25%), PrTX-III (35%) and MjTX-II (10%) on the mouse paw. CK levels evoked by isolated Asp49 or Lys49-PLA(2)s were reduced by 40% to 54% in the presence of CA and, in all cases, the membrane damaging activity of the toxins was also reduced. Circular dichroism spectra of the PLA(2)s in the presence and absence of CA showed that there was not any detectable secondary structural modification due to association between CA and the myotoxins. However, Fourier Transformed Infrared (FT-IR) analysis indicated that ionic and hydrophobic contacts contributed to stabilize this interaction.

Animals↗
PubMed2005

Presence of gular and parietal pits in Atretium schistosum (Serpentes, Colubridae), a singular trait not exclusive to psammophine snakes.

The sporadic occurrence of localised pits on parietal plates was recently discovered in different colubrid genera of the subfamily Psammophiinae; these were considered to play a role in sensory perception. In the present study, we describe the presence of similar structures in Atretium schistosum, another colubrid snake reportedly not belonging to the Psammophiinae. As this species is suspected of being phylogenetically distantly related to psammophine snakes, some hypotheses are provided to explain (1) the putative function of these pits, (2) their sporadic occurrence, and (3) to suggest when they may have evolved in the colubroid snake clade.

Animals↗
PubMed2005

Organochlorine detection in the shed skins of snakes.

Lizards and snakes eliminate heavy metals in their shed skins. There are no data with regard to reptilian skin as a depuration route for organochlorine (OC) compounds; however, birds deposit OCs in feathers. Corn snakes (Elaphe guttata) were therefore fed thawed mice that had been injected with a mixture of alpha-chlordane, Aroclor 1254 (a polychlorinated biphenyl (PCB) mixture), and lindane (gamma-hexachlorocyclohexane) at 2, 8, and 4 mg/kg, respectively. Feeding of contaminated mice occurred on the first weekly feeding of each month, with remaining weekly feedings consisting of noncontaminated mice, and was continued for 6 months. Shed skins were evaluated in a multiresidue OC scan by gas chromatography. All three chemical contaminants were readily detected in the shed skins: chlordane, 0.155-0.213 ppm; PCB, 3.49-7.01 ppm; lindane: 0.028-0.042 ppm. These data suggest that the shed skin of snakes may serve as an elimination route for OC contaminants and as such may have utility as a noninvasive, nondestructive indicator tissue for assessing environmental contamination.

Animal Feed↗