Novo método de fracionamento da peçonha do escorpião Tityus serrulatus e caracterização eletrofisiológica das toxinas Ts6 e Ts7 / New fractionation procedure of Tityus serrulatus venom and electrophysiological characterization of toxins Ts6 and Ts7

AUTOR(ES)
FONTE

IBICT - Instituto Brasileiro de Informação em Ciência e Tecnologia

DATA DE PUBLICAÇÃO

01/06/2012

RESUMO

In Brazil, Tityus serrulatus (Ts) species is the responsible for the most scorpion accidents and also for the major incidence of accidents caused by venomous animals. About 16 different toxins of Ts venom have been listed so far, being Ts1 the major one. Among these toxins, the neurotoxins with action on sodium and potassium channels are the most interest in the scientific community, due to their effect in the envenomation and ion channel specificity. The neurotoxins with action on potassium channels are composed of an ?-helix and three ?-strands formed by 23-43 amino acid residues. They are classified into four families (?-, ?-, g- and ?- KTx). The ?-KTx family is the most relevant and is divided into 21 subfamilies. These toxins with high specificity for different subtypes of sodium and potassium channels are very important, because they can be used as therapeutic tools to specific target cells. Until now, the fractionation of these toxins was done using a CM-Cellulose-52 column, according to the protocol of Arantes and co-workers (1989). The present work standardized a new method of isolation using the same column, but incorporated the use of HPLC (High Performance Liquid Chromatography), in which was observed a chromatographic profile such as the previous one (XIII Fractions), however with high resolution and more practical. Some fraction of the previous method were divided in two subfractions (VIA-VIB, VIIIA-VIIIB, IXA-IXB e XIA-XIB), showing that the new method also present high resolution. Using the new method, it was isolated the Ts6 and Ts7 toxin. The effects of these toxins were evaluated in 14 different types of potassium channels (Kv1.1, Kv1.2, Kv1.3, Kv1.4, Kv1.5, Kv1.6, Kv2.1, Kv3.1, Kv4.3, Kv7.1, Kv7.2, Kv7.4, hERG and Shaker), which were expressed in Xenopus laevis oocytes using the voltage-clamp technique with twomicroelectrodes. The Ts6 toxin (1?M) shows to act on 11 types of potassium channels (Kv1.1, Kv1.2, Kv1.3, Kv1.5, Kv1.6, Kv4.3, Kv7.1, Kv7.2, Kv7.4, hERG and Shaker), but the blocking of Kv1.2 and Kv1.3 was significantly more intense. Using dose-response experiments, it was possible to confirm this selectivity, in which Ts6 demonstrates to act in both channels in extremely low quantities (IC50 Kv1.2 = 6,19 ± 0,35 nM /IC50 Kv1.3 = 0,55 ± 0,20 nM). The Ts7 toxin (1?M) shows to act on 11 types of potassium channels (Kv1.1, Kv1.2, Kv1.3, Kv1.5, Kv1.6, Kv4.3, Kv7.1, Kv7.2, Kv7.4, hERG and Shaker), but the blocking action on multiple subtypes channels showed no significant differences, showing low selectivity among the channels analyzed. This work was important to improve and facilitate the method of fractionation of Ts venom, as well as evaluate the electrophysiology properties of the toxins Ts6 and Ts7 of interacting with different types of potassium channels. These studies will be essential for future applications of these toxins as drugs to treat channelpathies or as tools to study potassium channels structurally and functionally.

ASSUNTO(S)

?-ktx a-ktx canais para potássio electrophysiology eletrofisiologia fracionamento de toxinas fractionation of toxins neurotoxinas neurotoxins potassium channels tityus serrulatus tityus serrulatus ts6 ts6 ts7 ts7

ACESSO AO ARTIGO

Documentos Relacionados