Wang Kun, Yin Shi-Jin, Lu Meng, Yi Hong, Dai Chao, Xu Xiu-Jing, Cao Zhi-Jian, Wu Ying-Liang, Li Wen-Xin
State Key Laboratory of Virology, College of Life Sciences, Wuhan University, Wuhan, Hubei 430072, P.R. China.
Biotechnol Lett. 2006 Nov;28(21):1767-72. doi: 10.1007/s10529-006-9155-y. Epub 2006 Aug 16.
The gene encoding the BmalphaTX14 (alpha-neurotoxin TX14) protein, derived from the cDNA library of the Chinese scorpion Buthus martensii Karsch, was expressed in Pichia pastoris. The recombinant protein was purified by metal chelate affinity chromatography and gel filtration chromatography. Using patch-clamp technique, electrophysiological activity of rBmalphaTX14 was identified. In the neurons isolated from mice trigeminal root ganglion, the Na+ current amplitude was reduced by 80% under whole cell patch-clamp recording. There were no apparent modifications to the gating mechanism in the presence of rBmalphaTX14. Although BmalphaTX14 shared a high amino acid sequence similarity with other typical alpha-toxins, it has different effects on neurons. Further electrophysiological analysis suggested that rBmalphaTX14 selectively blocked Na+ channels and is a member of a new group of scorpion toxins.
从中国蝎子东亚钳蝎的cDNA文库中获得的编码BmalphaTX14（α-神经毒素TX14）蛋白的基因，在毕赤酵母中表达。重组蛋白通过金属螯合亲和层析和凝胶过滤层析进行纯化。使用膜片钳技术鉴定了rBmalphaTX14的电生理活性。在从小鼠三叉神经根神经节分离的神经元中，在全细胞膜片钳记录下，Na+电流幅度降低了80%。在存在rBmalphaTX14的情况下，门控机制没有明显改变。尽管BmalphaTX14与其他典型的α-毒素具有高度的氨基酸序列相似性，但它对神经元有不同的影响。进一步的电生理分析表明，rBmalphaTX14选择性地阻断Na+通道，是一类新的蝎子毒素成员。
