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At least 973 records · Page 54Linked to original sources
PubMed2001

IsCT, a novel cytotoxic linear peptide from scorpion Opisthacanthus madagascariensis.

A novel cytotoxic linear peptide, IsCT, was characterized from scorpion Opisthacanthus madagascariensis. It is a linear peptide with a molecular weight of 1501.9 Da composed of 13 amino acid residues without cysteines. MS/MS analysis showed that its C-terminal is amidated. The identity of IsCT is re-confirmed by comparing the chemical synthesized peptide with the natural one. IsCT demonstrated antimicrobial activity against both gram-positive and gram-negative bacteria and hemolytic activity to sheep red blood cells. Also, it can release histamine from rat peritoneal mast cells. The CD absorption suggested that IsCT had an alpha-helix configuration in aqueous TFE. IsCT is one of the shortest natural cytotoxic peptides described, and it will be a suitable model for studying peptide-lipid interactions.

Amino Acid Sequence↗
PubMed2001

Automatic assignment of NOESY cross peaks and determination of the protein structure of a new world scorpion neurotoxin using NOAH/DIAMOD.

The 3D NMR structures of the scorpion neurotoxin, CsE-v5, were determined from the same NOESY spectra with NOAH/DIAMOD, an automated assignment and 3D structure calculation software package, and with a conventional manual assignment combined with a distance geometry/simulated annealing (X-PLOR) refinement method. The NOESY assignments and the 3D structures obtained from the two independent methods were compared in detail. The NOAH/DIAMOD program suite uses feedback filtering and self-correcting distance geometry methods to automatically assign NOESY spectra and to calculate the 3D structure of a protein. NOESY cross peaks were automatically picked using a standard software package and combined with 74 manually assigned NOESY peaks to start the NOAH/DIAMOD calculations. After 63 NOAH/DIAMOD cycles, using REDAC procedures in the last 8 cycles, and final FANTOM constrained energy minimization, a bundle of 20 structures with the smallest target functions has a RMSD of 0.81 A for backbone atoms and 1.11 A for all heavy atoms to the mean structure. Despite some missing chemical shifts of side chain protons, 776 (including 74 manually assigned) of 1130 NOE peaks were unambiguously assigned, 150 peaks have more than one possible assignment compatible with the bundle structures, and only 30 peaks could not be assigned within the given chemical shift tolerance ranges in either the D1 or the D2 dimension. The remaining 174, mainly weak NOE peaks were not compatible with the final 20 best bundle structures at the last NOAH/DIAMOD cycle. The automatically determined structures agree well with the structures determined independently using the conventional method and the same NMR spectra, with the mean RMSD in well-defined regions of 0.84 A for bb and 1.48 A for all heavy atoms from residues 2-5, 18-26, 32-36, and 39-45. This study demonstrates the potential of the NOAH/DIAMOD program suite to automatically assign NMR data for proteins and determine their structure.

Amino Acids↗
PubMedDate not supplied

Three-dimensional immunoelectron microscopy of scorpion hemocyanin labeled with a monoclonal Fab fragment.

An immunocomplex of the 4 x 6-meric hemocyanin of the scorpion Androctonus australis with the monoclonal Fab fragment L104 was reconstructed from electron micrographs of a negatively stained specimen, using the double-carbon-layer technique. The resulting structure enables a clear visualization of the Fab fragments bound to the four copies of the Aa6 subunit and directly confirms a previous localization of the L104 epitope deduced from two-dimensional image processing. Despite a strong flattening effect produced by the negative-staining technique the orientations of the Fab fragments are well characterized. Moreover, the observation of a central hole within the elbow bends of the Fab fragments provides information about the disposition of the Fabs around their main axis.

Animals↗
PubMed1999

Expression and purification of the BmK M1 neurotoxin from the scorpion Buthus martensii Karsch.

The gene encoding a neurotoxin (BmK M1) from the scorpion Buthus martensii Karsch was expressed in Saccharomyces cerevisiae at a high level with the alcohol dehydrogenase promoter. SDS-PAGE of the culture confirmed expression and showed secretion into medium from yeast. Recombinant BmK M1 was purified rapidly and efficiently by ion exchange and gel filtration chromatography to homogeneity, produced a single band on tricine-SDS-PAGE, and processed the homologous N-terminus. Amino acid analysis and N-terminal sequencing demonstrated that the recombinant toxin was processed correctly from the alpha-mating factor leader sequence and was chemically identical to the native form. The expressed recombinant BmK M1 was toxic for mice, which indicated that it was biologically active. Quantitative estimation showed that recombinant BmK M1 had an LD(50) similar to that of the native toxin.

Animals↗
PubMed1992

Isolation of a bradykinin-potentiating factor from scorpion Tityus serrulatus venom.

A bradykinin-potentiating factor was isolated and characterized from the scorpion Tityus serrulatus venom by chromatographic techniques and reverse phase followed by biological assays. This factor showed to be able to potentiate the contractile activity of the isolated guinea-pig ileum, inhibited the angiotensin-converting enzyme and potentiated the bradykinin-induced lowering of the arterial blood pressure in the rat.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1994

Neurogenic chloride secretion induced by scorpion venom and veratrine in rabbit colon.

In earlier reconstitution experiments the venom of the scorpion Leiurus quinquestriatus, LQV, was shown to block Ca(2+)-activated high-conductance K+ channels from the basolateral cell membrane of rabbit colonocytes (Turnheim K, Costantin J, Chan S, Schultz SG (1989) J Membrane Biol 112:247-254). These LQV-sensitive K+ channels do not seem to be involved in active Na+ transport across rabbit colon, as absorptive Na+ fluxes were not significantly affected by serosal addition of LQV to isolated epithelia of rabbit descending colon. While Na+ absorption was not changed, LQV and veratrine caused electrogenic Cl- secretion in this tissue by a neural (tetrodotoxin sensitive) mechanism. The secretory effect of LQV was partly inhibited by atropine, suggesting the involvement of m-cholinoceptors, and by a VIP-antagonist. In contrast to the neurogenic secretion in the small intestine of guinea pig, rat and cat, 5-hydroxytryptamine (5-HT) does not seem to be involved in neurogenic secretion in rabbit colon, as 1) several 5-HT receptor antagonists did not inhibit the LQV effect with the exception of high concentrations of tropisetron, 2) exogenous 5-HT had no secretory effect, and 3) there was no significant release of 5-HT from the tissue during neurogenic secretion. The inhibitory effect of tropisetron on intestinal Cl- secretion seems to be unrelated to its property as a 5-HT3 receptor antagonist.

Animals↗
PubMed1982

Neurosecretory cells in the circadian-clock system of the scorpion, Androctonus australis.

The somata of the efferent neurosecretory fibers that control the circadian sensitivity rhythm in the median eyes of the scorpion, Androctonus australis, were detected in the brain by retrograde labeling with Lucifer Yellow CH. A total of 20-40 neurons are arranged in two groups displaying a bilaterally symmetrical, marginal position near the circumesophageal connectives. Half the cells in each group send fibers into the ipsilateral optic nerve; the fibers from the other half enter the contralateral optic nerve.

Animals↗
PubMed1977

Neursecretory fibres in the median eyes of the scorpion, Androctonus australis L.

The retina of the median eyes of the North African scorpion, Androctonus australis L., is supplied with numerous neurosecretory nerve fibres which establish synaptoid contacts on the retinula cells. The number of fibres or profiles of varicosities of fibre terminals associated with a retinular unit (five retinula cells with a fused rhabdom) varies between 10 and 20. Electron-opaque vesicles with a diameter of 80-100 nm are abundant within the axonal profiles. The synaptoid junctions are characterized by postsynaptic electron-dense material on the inner leaflet of the retinula cell membrane and, frequently, presynaptic submembranous dense material. Because of these ultrastructural features, the junctions observed here resemble typical interneuronal synaptic contacts. Hence this kind of neurosecretory junction appears to be unique among arthropods. It is suggested that the neurosecretory fibres within the retina represent the efferent pathways for the control of the circadian pigment movements within the retinula cells.

Animals↗
PubMed1980

The lateral eyes of the scorpion, Androctonus australis.

The dioptric apparatus of the lateral eyes of the scorpion, Androctonus australis, consists of a cuticular lens, but lacks a vitreous body. The retina is formed by (1) retinula cells displaying a contiguous network of rhabdoms; (2) arhabdomeric cells bearing a distal dendrite that contacts retinula cells via numerous projections and ends before the rhabdomere of the retinula cells; (3) pigment cells that ensheath retinula and arhabdomeric cells with the exception of the contact regions; and (4) neurosecretory fibres possibly originating in the supraesophageal ganglion. The ratio of the number of retinula to arhabdomeric cells is determined to be close to 2:1 in the three larger anterolateral eyes, in contrast to the median eyes where the ratio is 5:1. The construction of the dioptric apparatus as well as the anatomy of the retina imply that in the lateral eyes of Androctonus australis visual acuity is reduced. A certain degree of spatial discrimination, however, may be retained by the presence of a relatively high number of arhabdomeric cells. It is suggested that the lateral eyes of A. australis mainly function as light detectors, e.g., for Zeitgeber stimuli.

Animals↗
PubMed1989

Effects of toxin II from the scorpion Androctonus australis Hector on sodium current in neuroblastoma cells and their modulation by oleic acid.

The effects of toxin II (AaH II) isolated from the scorpion Androctonus australis Hector on sodium current in neuroblastoma X glioma NG 108-15 hybrid cells were analysed under patch clamp conditions in the whole cell configuration. AaH II (70 nM) induced a maintained sodium current, as well as increasing both fast and slow inactivation time constants and the amplitude of the peak current. This latter effect occurred via a shift of the activation-voltage curve towards negative voltage values by about 9 mV. Oleic acid (5 microM), which had no effect on INa under control conditions, decreased the AaH II-induced maintained current. It also reversed, or prevented the increase of the peak current induced by AaH II. However, it neither prevented nor modified the AaH II-induced increase in inactivation time constants. The binding of the toxin to its specific site and the number of binding sites for AaH II were not significantly modified by oleic acid. The oleic acid-induced effects could not be related to the activation of protein kinase C since PMA, a potent activator of this enzyme, did not produce oleic acid-like effects. From these results, it is concluded that AaH II has several independent effects on sodium channels, some of which could be modulated by the lipid environment of sodium channels in the membrane.

Animals↗
PubMed1989

Changes in Na channel properties of frog and rat skeletal muscles induced by the AaH II toxin from the scorpion Androctonus australis.

The effects of the mammal toxin II isolated from the venom of the scorpion Androctonus australis Hector (AaH II) were studied under current and voltage clamp conditions in frog (semitendinosus) and rat (fast e.d.l. and slow soleus) skeletal twitch muscle fibres. In both species, AaH II induced a dose-dependent prolongation of the action potential (AP) leading at saturating concentration to APs with long plateaus of about 1.5 s in frog and 5 s in rat e.d.l. and soleus fibres. The concentrations to induce 50% of the maximal effect (K0.5) were 9.1 x 10(-9) M in the frog and 1.4 x 10(-9) M in the rat. AaH II increased the time constants of inactivation of the peak Na current and induced a maintained Na current that was greater in rat e.d.l. and soleus (31.6% of peak current amplitude at -30 mV; K0.5 = 0.8 x 10(-9) M) than in frog (16.5%; K0.5 = 15.5 x 10(-9) M) muscles. Peak and maintained Na currents were TTX-sensitive and had identical threshold and reversal potentials. The half-maximum maintained permeability occurred at a potential 20 mV more positive than the peak permeability. Recovery from inactivation and steady-state inactivation of the inactivating Na current remained unchanged. The maintained current deactivated with normal fast kinetics. The action of the toxin reversed poorly on washout but could be largely removed by conditioning depolarizations more positive than the reversal potential of the Na current. Our results suggest that, in vertebrate skeletal muscle fibres, AaH II affects all the Na channels and are consistent with the hypothesis that the maintained current originates from a reopening of previously inactivated Na channels.

Action Potentials↗
PubMed1975

Pharmacology of scorpion toxin II in the skeletal muscle.

1. Scorpion toxin II is potent in inducing contracture and spontaneous contractions of the chick biventer cervicis muscle. In addition, this toxin induces membrane depolarization and blockade of neuromuscular transmission in this muscle preparation. The purpose of the present study is to explore the possible mechanism of actions of toxin II. 2. The muscle contracture induced by toxin II is moderately accelerated by Ca2+-free Krebs solution, delayed by high Ca2+ (10 mM), high Mg2+ (10 mM) and low Na+ (60mM) Krebs solution. Moreover, this action is inhibited slightly by d-tubocurarine and completely by either procaine or tetrodotoxin, but unaffected by beta-bungarotoxin. All these findings suggest that toxin II induces contracture mainly by increasing the Na+ permeability of the muscle membrane. 3. Spontaneous contractions induced by toxin II are abolished by Ca2+-free Krebs solution, inhibited partially by either d-tubocurarine or beta-bungarotoxin and completely by tetrodotoxin or procaine. These results suggest that toxin II induces spontaneous contractions partially by releasing acetylcholine from nerve endings and partially by increasing the Na+ permeability of the muscle membrane.

Acetylcholine↗
PubMed1986

Toxin gamma of the scorpion Tityus serrulatus modifies both activation and inactivation of sodium permeability of nerve membrane.

The effects of the major neurotoxic fraction isolated from scorpion venom of Tityus serrulatus, TiTx gamma, on peripheral nerve membrane of Xenopus laevis were studied under current- and voltage-clamp conditions. 700 nmol/l TiTx gamma depolarized the membrane and induced spontaneous activity (150 s-1, maximum value), which ceased within a few minutes. It reduced the amplitude of the action potentials from 109 mV to 52 mV and increased their duration from 1.25 ms to 4.5 ms. 440 nmol/l TiTx gamma induced inward Na current flow at resting potential. The descending branch of the Na current-voltage curve was flattened and shifted approximately 10 mV to more negative potentials. Maximum Na permeability was reduced to about 20%. Both development of and recovery from inactivation of Na permeability were slowed. The steepness of the steady-state inactivation curve was decreased, but the mid-potential changed only insignificantly. No prepulse was necessary to elicit either a shift of activation or an inward current at resting potential. Expressing the toxin effect either in terms of the decrease of Na peak current or of the slowing of inactivation, half-maximum effects were found with 0.3 +/- 0.1 and 3.7 +/- 0.7 mumol/l TiTx gamma, respectively.

Action Potentials↗
PubMed1981

Potential-dependent effects of sea anemone toxins and scorpion venom on crayfish giant axon.

Effects of two kinds of sea anemone toxin (Parasicyonis actinostoloides and Anemonia sulcata) and scorpion venom (Leiurus quinquestriatus) on crayfish giant axons were examined electrophysiologically. All toxins acted on the axon in a similar manner to prolong the falling phase of the action potential. In all cases the development of toxicity was reduced when the nerve membrane was depolarized by a current injection. However, the ranges of membrane potential where the significant reduction in toxicity took place were different for each toxin. The action of Parasicyonis toxin was also suppressed by depolarization resulting from treatment of the axon with a neurotoxic alkaloid, veratridine. The mechanism of the potential-dependent toxin action is discussed with reference to the present data and relevant works by other investigators.

Animals↗
PubMed1982

Effect of toxins isolated from the venom of the scorpion Centruroides sculpturatus on the Na currents of the node of Ranvier.

1. The effect of various toxin fractions isolated by Watt et al. (1978) from the venom of the scorpion Centruroides sculpturatus Ewing on the Na currents of the node of Ranvier has been studied with the voltage clamp method. 2. The toxin fractions were applied externally. The most potent fractions were toxins III, IV and V which were effective in concentrations of 0.33-3.33 microgram/ml. The effect of toxins III and IV was quite different from that of toxin V. 3. In toxin III or IV - treated nodes a strong depolarizing pulse was followed by a transient shift of the negative resistance branch of the INa (E) curve to more negative potentials. The amount of shift varied between -10 and -60 mV. A 500 ms depolarizing pulse of small amplitude produced a slowly developing Na inward current which slowly decayed after the end of the pulse. Inactivation was incomplete, even with 500 ms pulses to 0 mV. 4. The transient shift of the INa (E) curve was not seen in nodes treated with toxin V. This toxin merely caused slow and incomplete Na inactivation. The effect of toxin IV was not suppressed by a four times higher concentration of toxin V, suggesting that the two toxins act on different receptors. 5. Toxin I acted like toxin IV but was about 10 times less potent. The effect of high concentrations of variants 1, 2, 3, 5, 6 resembled tha of toxin V. 6. All effects observed with toxin III or IV were also seen with the whole venom (cf. Cahalan 1975).

Animals↗
PubMed1984

The effect of Tityus serrulatus scorpion toxin gamma on Na channels in neuroblastoma cells.

The effects of highly purified toxin gamma from the venom of the scorpion Tityus serrulatus (TiTx gamma) on nerve membrane ionic channels have been investigated using the suction electrodes voltage clamp technique on neuroblastoma cells. The amplitude of the normally voltage-dependent Na current is reversible reduced by approximately 50% after 15-105 nM TiTx gamma, whereas even the highest toxin concentrations have no significant effect on the outward K current in the presence of tetrodotoxin. TiTx gamma causes a transient inward current to appear at membrane potentials between -70 and -40 mV, a potential region in which no significant inward current is observed in control experiments. Tetrodotoxin (300 nM) rapidly blocks both the TiTx gamma-induced inward current and the remaining normally voltage-dependent Na current. The binding of radiolabelled TiTx gamma to the Na channels in the neuroblastoma cell membrane is prevented by native TiTx gamma with a K0.5 = 0.75 nM. Both activation and inactivation of the TiTx gamma-induced Na current are shifted 30-40 mV towards more negative potential values as compared to normally voltage-dependent Na current. The TiTx gamma-induced Na current exhibits sigmoidal activation kinetics and relatively slow, exponential inactivation kinetics. The local anesthetic procaine at an external concentration of 1 mM blocks more effectively the remaining normally voltage-dependent Na current than the TiTx gamma-induced Na current. Both Na current components are equally blocked by 1 mM of the local anesthetic propoxycaine. The relation between the effects of TiTx gamma on Nat channels and those of other known neurotoxins those of other known neurotoxins specific of this channel is discussed.(ABSTRACT TRUNCATED AT 250 WORDS)

Anesthetics, Local↗
PubMed1986

Effects of toxins VI and VII from the scorpion Centruroides sculpturatus on the Na currents of the frog node of Ranvier.

New toxins, VI and VII, were purified from the venom of the North American scorpion, Centruroides sculpturatus, and studied in the voltage-clamped frog node of Ranvier. These toxins reduced peak inward Na currents and caused a transient, depolarization-induced shift in the voltage dependence of Na activation. Their effects were indistinguishable from those of toxins I, III and IV, as previously described by Meves et al. (1982). Toxin VII, 0.2 microgram/ml, shifted the voltage dependence of steady-state inactivation (h infinity). In four fibers, the mean shift of the h infinity (E) curve was -17 mV, compared to a mean shift of -28 mV in the descending branch of the INa(E) curve. The h infinity (E) curve with toxin VII was monotonic and inactivation was incomplete at positive potentials. Decreasing the pH from 7.4 to 5.7 increased the shift in the voltage dependence of activation with toxin. The increase with toxin VI at pH 5.7 was reversible on returning to pH 7.4, but the increase with toxin VII was not. The lack of reversibility of the effect of pH with toxin VII was quantified in two ways: the mean value of INa (measured at -62 mV with a conditioning pulse) at pH 7.4 was 5.1 times larger after treatment than before treatment with 0.1 microgram/ml of toxin VII at pH 5.7; the average concentration of toxin VII required for a shift of -30 mV at pH 7.4 was decreased by a factor of 4.3, to 0.19 microgram/ml (26 nM), by pretreatment at pH 5.7.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Voltage-dependent effect of a scorpion toxin on sodium current inactivation.

In voltage clamped nodes of Ranvier inactivation of the sodium permeability is slowed by toxin V from the scorpion Centruroides sculpturatus, by sea anemone toxin ATX II or by internally applied KIO3. The slow decay of the Na inward current is markedly accelerated if the test pulse is preceded by a depolarizing conditioning pulse followed by a 10-500 ms pause. This phenomenon was studied in detail, using conditioning pulses of varying amplitude and up to 15 s duration. In nodes treated with toxin V a 20 ms conditioning pulse to positive potentials was sufficient to produce a clear acceleration of the decay of the Na current and a reduction of the inward current remaining at the end of a 50 ms test pulse, i.e. a weakening of the toxin effect. In nodes treated with ATX II or internal KIO3 longer conditioning pulses were required. A similar effect of conditioning pulses on the decaying phase of the Na current was also observed in untreated fibres. To study the phenomenon quantitatively we fitted the decaying phase of the inward Na current with the equation INa = A exp(-t/tau 1) + B exp(-t/tau 2) + C. The effect of depolarizing conditioning pulses could be described as an increase of A, a decrease of B and C and a reduction of the time constants tau 1 and tau 2. I50/Ipeak, the normalised inward current remaining at the end of a 50 ms test pulse, decreased exponentially with increasing duration of the conditioning pulse to a steady-state value. The time constant tau and the steady-state value depended on the potential during the conditioning pulse. For nodes treated with toxin V, tau was 0.24 s at 0 mV and 12 degrees C and half inhibition occurred at -42 mV. The time constant tau was larger for nodes treated with ATX II or internal KIO3. At positive potentials, I50 was reduced to 20% of the control value in toxin V-treated nodes, but only to 70% in KIO3-treated nodes. Recovery from the effect of the conditioning pulse was studied by varying the pause between conditioning pulse and test pulse; recovery was 66-100% complete after 500 ms. The results are interpreted by assuming that a depolarizing conditioning pulse (a) accelerates inactivation of the sodium permeability and (b) causes dissociation of the toxin-receptor complex or transition into an inactive state. The latter effect occurs in toxin V-treated fibres but not in those treated with ATX II or KIO3.

Animals↗