Cloning and characterization of BmK86, a novel K+-channel blocker from scorpion venom
Cloning and characterization of BmK86, a novel K+-channel blocker from scorpion venom
复制标题
BmK86（一种来自蝎毒的新型 K 通道阻断剂）的克隆和表征。
DOI：
10.1016/j.bbrc.2007.06.108
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发表时间：
2007-09-07
影响因子：
3.1
通讯作者：
Li, Wenxin
中科院分区：
文献类型：
--
作者：
Mao, Xin;Cao, Zhijian;Li, Wenxin
蝎子毒液代表了一个巨大的迄今未开发的资源，以了解离子通道。BmK86是从马氏Mesobuthus martensii Karsch的cDNA文库中分离到的一个新的K+通道毒素基因，编码一个包含22个氨基酸残基的信号肽和一个包含35个残基的含3个二硫桥的成熟毒素。BmK86的基因组序列由两个被72bp的内含子破坏的外显子组成。与其他蝎子毒素相比，BmK86序列相似性较低。GST-BmK86融合蛋白在大肠杆菌中成功表达。融合蛋白经肠激酶裂解，重组BmK86经高效液相色谱纯化。利用全细胞膜片钳记录，发现重组BmK86能抑制COS7细胞中mKv1.3通道钾电流的表达。这些结果表明，BmK86属于一个新的α - ktxs亚家族的代表成员。BmK86的系统编号为alpha-KTx26.1。(c) 2007爱思唯尔公司版权所有。
Scorpion venom represents a tremendous hitherto unexplored resource for understanding ion channels. BmK86 is a novel K+-channel toxin gene isolated from a cDNA library of Mesobuthus martensii Karsch, which encodes a signal peptide of 22 amino acid residues and a mature toxin of 35 residues with three disulfide bridges. The genomic sequence of BmK86 consists of two exons disrupted by an intron of 72 bp. Comparison with the other scorpion toxins BmK86 shows low sequence similarity. The GST-BmK86 fusion protein was successfully expressed in Escherichia coli. The fusion protein was cleaved by enterokinase and the recombinant BmK86 was purified by HPLC. Using whole-cell patch-clamp recording, the recombinant BmK86 was found to inhibit the potassium current of mKv1.3 channel expressed in COS7 cells. These results indicated that BmK86 belongs to a representative member of a novel subfamily of alpha-KTxs. The systematic number assigned to BmK86 is alpha-KTx26.1. (c) 2007 Elsevier Inc. All rights reserved.