Abstract

A technique has been developed for obtaining recombinant functionally active peptides Ce1 and Ce4 from the venom of the scorpion Centruroides elegans in the Escherichia coli expression system. The yields of peptides Ce1 and Ce4 were 6.5 and 12 mg per liter of culture, respectively. The properties of the obtained peptides were studied using bioengineered systems based on hybrid channels KcsA-K_v1.x (x = 1, 3, 6) containing blocker binding sites of the corresponding eukaryotic potassium channels of K_v1-family. It has been shown that recombinant Ce1 and Ce4 do not exhibit affinity to the binding sites of K_v1.1 and K_v1.6 channels up to micromolar concentrations and, like natural peptides, selectively interact with the binding site of the K_v1.3 channel: the apparent dissociation constants of KcsA-K_v1.3 complexes with recombinant Ce1 and Ce4 are 50 ± 10 and 200 ± 30 nM (mean ± SEM), respectively.

中文翻译：

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来自 Scorpion Centruroides elegans 毒液的重组肽 Ce1 和 Ce4 及其与混合通道 KcsA-Kv1.x (x = 1, 3, 6) 的相互作用

摘要

已经开发出一种技术，用于从大肠杆菌表达系统中的线虫蝎毒液中获得重组功能活性肽 Ce1 和 Ce4。肽 Ce1 和 Ce4 的产量分别为每升培养物 6.5 和 12 mg。使用基于混合通道 KcsA-K _{v 1.x (x = 1, 3, 6) 的生物工程系统研究获得的肽的特性，该混合通道包含 K v} 1 家族的相应真核钾通道的阻断剂结合位点。_{已经表明，重组 Ce1 和 Ce4 对 K v} 1.1 和 K _v的结合位点没有亲和力1.6 通道高达微摩尔浓度，并且像天然肽一样，选择性地与 K _v 1.3 通道的结合位点相互作用：KcsA-K _v 1.3 与重组 Ce1 和 Ce4 复合物的表观解离常数为 50 ± 10 和 200 ± 30 nM （平均值±SEM），分别。