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A Bdolah

Publications and source records attributed to A Bdolah.

At least 37 records · Page 2Linked to original sources
PubMed1990

Different pathways of endothelin/sarafotoxin-stimulated phosphoinositide hydrolysis in myocytes.

Aging of rat heart myocytes in culture is accompanied by approximately 50% reduction in endothelin (ET)/sarafotoxin (SRTX) receptor-binding capacity as well as in the induction of phosphoinositide (PI) hydrolysis. Treatment of aged cultures under conditions yielding myocytes with a lipid composition similar to that in young cultures restored all the ET/SRTX receptors; at the same time it re-established only the endothelin-induced but not the sarafotoxin-induced PI-hydrolysis response. Thus more than one mechanism may stimulate PI metabolism.

Animals↗
PubMed1990

Endothelins are more sensitive than sarafotoxins to neutral endopeptidase: possible physiological significance.

Incubation of endothelins (ETs) with bovine kidney neutral endopeptidase (NEP) resulted in a selective two-step degradation with loss of biochemical activity. The Km of the enzyme indicated high-affinity binding, and hydrolysis was completely inhibited by phosphoramidon. The first step was nicking of the Ser5-Leu6 bond, followed by cleavage at the amino side of Ile19. The nicked peptide exhibited biochemical activities comparable to those of the intact peptide--i.e., binding to the ET receptor, induction of inositol phospholipid hydrolysis, and toxicity. The twice-cleaved product was inactive. The sarafotoxins (SRTXs) were more resistant than the ETs to NEP: for example, the half-time for ET-1 was approximately 1 hr, while it was approximately 4 hr for SRTX-b and even higher for SRTX-c. These in vitro findings may indicate a regulatory role of NEP (or similar enzymes) in the physiological inactivation of ETs. They might also help to explain why under certain physiological conditions ETs may be less toxic than SRTXs.

Amino Acid Sequence↗
PubMed1989

SRTX-d, a new native peptide of the endothelin/sarafotoxin family.

The primary structure of a new sarafotoxin, SRTX-d, from the venom of Atractaspis engaddensis is described. SRTX-d differs from SRTX-b in two substitutions: Ile19 instead of Val and Thr2 instead of Ser. The toxicity of SRTX-d and its vasoconstriction potency are very low in comparison to SRTX-a and SRTX-b, whereas its IC50 for 125I-SRTX-b binding is similar to that of SRTX-b. It is suggested that the Thr to Ser substitution, which is shared by two additional weak members of the endothelin/sarafotoxin family, SRTX-c and ET-3, affects the biological activity of SRTX-d as well.

Amino Acid Sequence↗
PubMed1989

Functional endothelin/sarafotoxin receptors in rat heart myocytes: structure-activity relationships and receptor subtypes.

Functional receptors for the peptides of the endothelin (ET) and sarafotoxin (SRTX) family were characterized in newborn rat heart myocytes using human and rat endothelins (ET-1 and ET-3, respectively), SRTX-b and SRTX-c. Binding studies in intact cells and homogenates revealed significantly higher affinities of ET-1 and SRTX-b than of ET-3 and SRTX-c towards these receptors. This binding profile of ET/SRTX peptides points to their interaction with the receptor subtype designated E-S alpha. All four peptides induced time- and dose-dependent phosphoinositide hydrolysis with the following rank order of potency: ET-1 greater than SRTX-b greater than SRTX-c greater than ET-3. Thus, ET-3 which possesses an intermediate affinity toward the receptor was the least effective with regard to this response. These results confirm and extend our earlier report that the ET/SRTX peptides interact with a newly characterized receptor(s) associated with phosphoinositide metabolism and Ca2+ mobilization. The initiation of inositol phosphate formation is largely independent of extracellular Ca2+, verapamil and nifedipine, indicating that the ET/SRTX peptides are not agonists for the voltage-dependent Ca2+-channels.

Animals↗
PubMed1989

Functional endothelin/sarafotoxin receptors in the rat uterus.

Functional receptors for the peptides of the endothelin (ET) and sarafotoxin (SRTX) families were detected in the rat uterus. These receptors specifically bind 125I-SRTX-b (Bmax = 220 fmol/mg protein), as well as ET-1, ET-3 and SRTX-c (IC50's 10, 5, 300 and 780 nM, respectively). Activation of the uterine ET/SRTX receptors induced dose-dependent phosphoinositide (PI) hydrolysis and three typical contractile responses: 1) increase in the muscle tonic tension; 2) increase in frequency of the spontaneous rhythmic contractions; 3) decrease of relaxation in each spontaneous rhythmic cycle. All three effects appeared at doses as low as 0.5-1 nM. Dose responses yield ED50 values of 5.5, 30 and 680 nM for ET-1, SRTX-b and ET-3, respectively. SRTX-c was the least effective peptide in achieving decrease in relaxation. In view of these results, and since the uterine responses to the peptides were almost immediate and reversible, we suggest that the functional ET/SRTX receptor of the rat uterus that is coupled to PI hydrolysis may be of physiological significance.

Animals↗
PubMed1989

Immunological and structural characterization of sarafotoxin/endothelin family of peptides.

A highly specific and sensitive radioimmunoassay (RIA) was developed for the potent vasoconstrictor peptides, sarafotoxin-b and human endothelin. The antigenic determinants of the antibodies employed in studies with these assays were found to be localized within the amino acid sequence at positions 4-7. This was confirmed by CNBr cleavage of the methionyl residue at position 6 in the sarafotoxin and at position 7 in the endothelin. The chemically characterized modified peptides showed very low cross reactivity in the RIAs. On the other hand, the binding properties as well as the ability to induce phosphoinositide hydrolysis were very similar in the modified and native peptides, indicating that despite cleavage of the peptide bond the biologically active conformation responsible for either binding or phosphoinositide hydrolysis is retained, probably because of the disulfide bonds. Thus, structural alteration might be a valuable means of curtailing some of the various activities induced by the sarafotoxin/endothelin family of peptides.

Animals↗
PubMed1989

Vasoconstrictor effects of sarafotoxins in rabbit aorta: structure-function relationships.

The sarafotoxins SRTX-a, b and c from the venom of the snake Atractaspis engaddensis are 21-amino acid peptides that affect the cardiovascular system. They are strong vasoconstrictors, the potency of which may be in correlation with their primary structure: SRTX-a, which differs from SRTX-b in a single amino acid residue (Asn instead of Tyr), shows about half of its maximal vasoconstriction, while SRTX-c, which differs in 3 additional residues is a very weak vasoconstrictor and, at high doses, shows vasodilatory effects. Sequential application of the three isotoxins result in a summated response.

Amino Acid Sequence↗
PubMed1989

Sarafotoxin receptors mediate phosphoinositide hydrolysis in various rat brain regions.

Sarafotoxin-b, a potent snake vasoconstrictor peptide homologous to the mammalian endothelial vasoconstrictor endothelin, induces phosphoinositide (PI) hydrolysis in various brain regions of the rat. Sarafotoxin-b induced PI hydrolysis is largely independent of extracellular Ca2+ and is detected in all brain regions where toxin-binding sites are found. These results point to the existence of a hitherto undetected neuroreceptor associated with the PI cycle.

Animals↗
PubMed1988

Characterization and localization of a novel neuroreceptor for the peptide sarafotoxin.

We have recently shown that the rat atrium and brain contain specific high affinity receptors for the novel snake vasoconstrictor peptide sarafotoxin-b (SRTXb), and demonstrated toxin-induced phosphoinositide hydrolysis. Here we report on the characteristics of 125I-SRTXb receptors and their regional distribution in rat brain. 125I-SRTX receptors in the rat brain bind the toxin rapidly and with high affinity. The binding was not inhibited by ligands of known neurotransmitter receptor and ion channels. 125I-SRTX receptors have a distinctive regional distribution. The highest densities were observed in the cerebellum, thalamus and hypothalamus (850, 550 and 450 fmol/mg protein, respectively) and the lowest densities in the caudate and cerebral cortex (82 and 62 fmol/mg protein, respectively). Taken together our results suggest that mammalian brains contain a hitherto undetected neuroreceptor that may operate in neurotransmission with a "SRTX-like" brain peptide, similar to the SRTX homologous vasoconstrictor peptide of the mammalian endothelium endothelin.

Animals↗
PubMed1988

Ascorbic acid and Mg-ATP co-regulate dopamine beta-monooxygenase activity in intact chromaffin granules.

Ascorbic acid and Mg-ATP were found to regulate norepinephrine biosynthesis in intact secretory vesicles synergistically and specifically, using the model system of isolated bovine chromaffin granules. Dopamine uptake into chromaffin granules was shown to be unrelated to the presence of Mg-ATP and ascorbic acid at external dopamine concentrations of 7.5 and 10 mM. Under these conditions of dopamine uptake, norepinephrine biosynthesis was enhanced 5-6-fold by Mg-ATP and ascorbic acid compared to control experiments with dopamine only. Furthermore, norepinephrine formation was enhanced approximately 3-fold by ascorbic acid and Mg-ATP together compared to norepinephrine formation in granules incubated with either substance alone. The action of Mg-ATP and ascorbic acid together was synergistic and independent of dopamine content of chromaffin granules as well as of dopamine uptake. The apparent Km of norepinephrine formation for external ascorbic acid was 376 microM and for external Mg-ATP was 132 microM, consistent with the larger amounts of cytosolic ascorbic acid and ATP that are available to chromaffin granules. Other physiologic reducing agents were not able to increase norepinephrine biosynthesis in the presence or absence of Mg-ATP. In addition, maximum enhancement of norepinephrine biosynthesis occurred only with the nucleotide ATP and the cation magnesium. The mechanism of the effect of ascorbic acid and Mg-ATP on norepinephrine biosynthesis was investigated and appeared to be independent of a positive membrane potential. The effect was also not mediated by direct action of ADP, ATP, or magnesium on the activity of soluble or particulate dopamine beta-monooxygenase. These data indicate that Mg-ATP and ascorbic acid specifically and synergistically co-regulate dopamine beta-monooxygenase activity in intact chromaffin granules, independent of substrate uptake. Although the mechanism is not known, the data are consistent with the possibility that the chromaffin granule ATPase mediates these effects.

Adenosine Triphosphate↗
PubMed1988

Sarafotoxin, a novel vasoconstrictor peptide: phosphoinositide hydrolysis in rat heart and brain.

Sarafotoxins, a group of 21-residue cardiotoxic peptides from snake venom that induce coronary vasoconstriction, show high-affinity binding to rat atrial and brain membranes and activate the hydrolysis of phosphoinositides. Neither their binding nor their activity is affected by blockers or activators of known receptors and ion channels, suggesting that sarafotoxins act either directly on the phosphoinositide phosphodiesterase system or on a novel receptor. Their amino acid sequence shows a high degree of homology with that of endothelin, a recently described 21-residue vasoconstrictor peptide found in porcine aortic endothelium. This is remarkable, since endothelin is a natural compound of the mammalian vascular system while sarafotoxins are highly toxic components of snake venom.

Amino Acid Sequence↗
PubMed1988

A novel cardiotoxic polypeptide from the venom of Atractaspis engaddensis (burrowing asp): cardiac effects in mice and isolated rat and human heart preparations.

A new cardiotoxic polypeptide isolated from the venom of the snake Atractaspis engaddensis has an LD50 of 15 micrograms/kg body weight in white mice. Intravenous administration in mice of lethal doses of the toxin causes, within seconds, marked changes in the ECG, consisting primarily of a transient slope elevation of the S-T segment, a temporary diminution of the S-wave and an increase in the amplitudes of the R- and T-waves. Concomitantly, and apparently unrelated to these changes, a severe A-V block develops and leads to complete cardiac arrest within a few min. Studies with rat and human isolated heart preparations showed that the toxin exerts a powerful coronary vasoconstriction (rats), and positive inotropic effects (rats and humans).

Animals↗
PubMed1987

Dietary regulation of digestive enzyme levels in the water snake, Natrix tessellata.

Levels of digestive enzymes were analyzed in water snakes following artificial feeding. A prominent increase of total proteolytic activity in the stomach was evident after feeding with a casein solution or after the snake was offered a fish (9- and 6-fold that of the fasting level, respectively). The activity following feeding with starch was much lower. Increased levels of chymotrypsin(ogen) as well as of amylase were evident in the pancreas 1 day after feeding the snake with fish. A specific induction of increased level of chymotrypsin in the pancreas of adult snakes was achieved by feeding with casein (12-fold that of the fasting level). In the group fed with starch, the chymotrypsin level dropped, while a 3-fold increase of amylase was evident. In newborn snakes, fed for the first time, casein induced a dramatic increase in the level of chymotrypsin in the pancreas (58 times the fasting level); feeding with starch induced an approximate 2-fold increase of chymotrypsin. Histological examination of the pancreas 1 day following casein feeding showed acinar cells loaded with zymogen granules. In starved animals and in snakes fed with starch, a much lower concentration of zymogen granules was observed. The pancreas of the snake may, thus, be most suitable for studying the specific induction of synthesis of digestive enzymes.

Acclimatization↗
PubMed1987

Interactions of the neurotoxic complex from the venom of the false horned viper (Pseudocerastes fieldi) with rat striatal synaptosomes.

The very high lethal potency of the neurotoxic complex (Cb) from the venom of Pseudocerastes fieldi following direct administration into the lateral ventricle of the brain, as compared with potency following i.v. administration, suggests that the toxin acts on the central nervous system. Rat striatal synaptosomes were selected to study interactions of the toxin with nerve endings. CbII, the toxic phospholipase A2 component of the toxin, as well as the reconstituted complex (CbI + CbII), inhibited the high affinity choline transport into synaptosomes. Fifty per cent inhibition was obtained at 10 nM CbII after 20 min preincubation of the synaptosomes at 37 degrees C. Choline uptake was inhibited under conditions of minimal leakage of lactate dehydrogenase and probably very low phospholipase A2 activity (in the absence of Ca2+ with Sr2+ or with EGTA). The inhibition of choline uptake was irreversible and was evident after a short preincubation at 0 degrees C. CbII also enhanced the release of acetylcholine from synaptosomes preloaded with labelled choline, but this effect was markedly reduced in the presence of the acidic component (CbI) of the complex. Binding of 125I-CbII could be demonstrated with synaptosomes and with erythrocytes, however, the reconstituted complex (CbI + CbII) was bound only by the synaptosomes, though less effectively than free 125I-CbII. An increased specific binding was evident with purified synaptosomes as compared with a crude preparation. These results support the notion that the non-toxic subunit increases the specificity of the toxic phospholipase A2.

Animals↗
PubMed1986

Gel permeation patterns of venoms from eleven species of the genus Vipera.

Gel filtration of venoms from eleven species of the genus Vipera shows similar patterns on Sephadex G-75, with three groups of protein peaks. The second group, which contains phospholipase A2, is variable among the species. As found in some other viperids, the venom of V. bornmuelleri shows synergism between phospholipase A and other fractions that considerably increases lethal potency.

Animals↗