The binding of the insect selective neurotoxin (AaIT) from scorpion venom to locust synaptosomal membranes |
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Authors: | Dalia Gordon Emmanuel Jover Francois Couraud Eliahu Zlotkin |
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Institution: | Laboratoire de Biochimie, INSERM U 172, Faculté de Médecine, Nord-Marseille France |
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Abstract: | The binding of the radioiodinated insect selective neurotoxin from the venom of the scorpion Androctonus australis (AaIT), to synaptic plasma membrane vesicles derived from osmotically shocked insect synaptosomes was studied under kinetic and equilibrium conditions. The integrity of these vesicles and the existence of membrane potential and its modifiability were demonstrated by assays of the uptake of the lipophilic cation tetraphenylphosphonium. It has been shown that 125I-labeled AaIT binds specifically and reversibly to a single class of noninteracting binding sites of high affinity () and low capacity (1.2–2.0 pmol/mg protein). The values of the rate association and dissociation constants k1 and k?1 are, respectively, 1.36 · 106 M?1 · s?1 and 1.9 · 10?3 s?1, and are in a good accordance with the equilibrium constant. The use of various ionophores and changes in external potassium concentration shown to modify the membrane potential of the present neuronal preparation, did not affect the binding of 125I-AaIT, thus indicating its voltage-independence. Veratridine, tetrodotoxin, sea anemone toxin and the α and β scorpion toxins specific for vertebrates did not affect the binding of 125I-AaIT. Furthermore, the above scorpion toxins were devoid of specific binding to the present insect neuronal preparation. Two additional insect toxins derived from the venom of the scorpion Buthotus judaicus, BjIT1 (spastic-excitatory toxin, homologus to the AaIT) and BjIT2 (flaccidity inducing-depressory toxin), were both shown to displace the 125I-AaIT with a high affinity (Kd = 2.2 and 1.3 nM, respectively). These data are compared and discussed in light of the information concerning the interaction of scorpion venom toxins affecting vertebrates with mammalian neuronal tissues. |
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Keywords: | Membrane-toxin interaction neurotoxin Insect selective toxin Membrane potential Synaptosome (Scorpion Locust) AaIT AmIT AaH II Css II Lqq V tetraphenyl phosphonium ion CCCP To whom correspondence should be addressed |
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