Three novel scorpion toxins, Aa1 from Androctonus australis, BmTX3 from Buthus martensi and AmmTX3 from Androctonus mauretanicus were shown able to selectively block A-type K+ currents in cerebellum granular cells or cultured striatum neurons from rat brain. In electrophysiology experiments, the transient A-current completely disappeared when 1 microM of the toxins was applied to the external solution whereas the sustained K+ current was unaffected. The three toxins shared high sequence homologies (more than 94%) and constituted a new 'short-chain' scorpion toxin subfamily: alpha-KTx15. Monoiododerivative of 125I-sBmTX3 specifically bound to rat brain synaptosomes. Under equilibrium binding conditions, maximum binding was 14 fmol/mg of protein and the dissociation constant (Kd) was 0.21 nM. This Kd value was confirmed by kinetic experiments (kon = 6.0 x 10(6) M(-1) s(-1) and koff = 6.0 x 10(-4) s(-1)). Competitions with AmmTX3 and Aa1 with 125I-sBmTX3 bound to its receptor on rat brain synaptosomes showed that they fully inhibited the 125I-sBmTX3 binding (Ki values of 20 and 44 pM, respectively), demonstrating unambiguously that the three molecules shared the same target in rat brain. A panel of toxins described as specific ligands for different K+, Na+ and Ca2+ channels were not able to displace 125I-sBmTX3 from its binding site. Thus, 125I-sBmTX3 is a new ligand for a still unidentified target in rat brain. In autoradiography, the distribution of 125I-sBmTX3 binding sites in the adult rat brain indicated a high density of 125I-sBmTX3 receptors in the striatum, hippocampus, superior colliculus, and cerebellum.
Definition of the alpha-KTx15 family.
2004
Abstract
Three novel scorpion toxins, Aa1 from Androctonus australis, BmTX3 from Buthus martensi and AmmTX3 from Androctonus mauretanicus were shown able to selectively block A-type K+ currents in cerebellum granular cells or cultured striatum neurons from rat brain. In electrophysiology experiments, the transient A-current completely disappeared when 1 microM of the toxins was applied to the external solution whereas the sustained K+ current was unaffected. The three toxins shared high sequence homologies (more than 94%) and constituted a new 'short-chain' scorpion toxin subfamily: alpha-KTx15. Monoiododerivative of 125I-sBmTX3 specifically bound to rat brain synaptosomes. Under equilibrium binding conditions, maximum binding was 14 fmol/mg of protein and the dissociation constant (Kd) was 0.21 nM. This Kd value was confirmed by kinetic experiments (kon = 6.0 x 10(6) M(-1) s(-1) and koff = 6.0 x 10(-4) s(-1)). Competitions with AmmTX3 and Aa1 with 125I-sBmTX3 bound to its receptor on rat brain synaptosomes showed that they fully inhibited the 125I-sBmTX3 binding (Ki values of 20 and 44 pM, respectively), demonstrating unambiguously that the three molecules shared the same target in rat brain. A panel of toxins described as specific ligands for different K+, Na+ and Ca2+ channels were not able to displace 125I-sBmTX3 from its binding site. Thus, 125I-sBmTX3 is a new ligand for a still unidentified target in rat brain. In autoradiography, the distribution of 125I-sBmTX3 binding sites in the adult rat brain indicated a high density of 125I-sBmTX3 receptors in the striatum, hippocampus, superior colliculus, and cerebellum.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
