| 德国小蠊钠离子通道V295N突变对α-蝎毒素LqhαIT敏感性的影响 |
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投稿时间：2024-08-31
修订日期：2025-01-06
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| 引用本文：温淑璇,马媛,陆容材,等.德国小蠊钠离子通道V295N突变对α-蝎毒素LqhαIT敏感性的影响[J].农药学学报,2025,27(1):90-97. |
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| 基金项目:三亚崖州湾菁英人才科技专项项目(SCKJ-JYRC-2023-15). |
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| 中文摘要:电压门控钠离子通道在细胞电信号传导中是必不可少的，它控制神经元和其他可兴奋细胞间动作电位的启动和传播，钠离子通道的快速失活过程是电信号传导的关键。蝎毒素作用于钠离子通道，影响通道的快速失活速度，因此解析蝎毒素与钠离子通道相互作用机制意义重大。通过比对德国小蠊(Blattella germanica)、美洲大蠊（Periplaneta americana）和哺乳动物等钠离子通道的氨基酸序列，发现4个差异氨基酸分别为I291L、Y292F、V295N和L406I，使用定点突变技术在德国小蠊钠离子通道上构建4种氨基酸突变质粒，以非洲爪蟾卵母细胞作为表达载体，经电生理检测发现V295N突变显著降低钠离子通道对α-蝎毒素LqhαIT的敏感性（约21.63倍）。进一步通过3D同源建模和蝎毒素分子对接发现，V295N突变使得钠离子通道M291与LqhαIT的K63位氨基酸形成了氢键，阻碍其与LqhαIT结合。综上，V295N突变间接影响钠离子通道与LqhαIT的结合，进而降低了钠离子通道对LqhαIT的敏感性。 |
| 中文关键词:德国小蠊 钠离子通道 LqhαIT 定点突变 双电极电压钳 同源建模 |
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| Influence of the V295N mutation of the Blattella germanica sodium channel on the sensitivity of the α-scorpion toxin LqhαIT |
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| Abstract:Voltage-gated sodium channels(VGSC) are essential in cell electrical signal transmission, which controls the initiation and propagation of action potentials between neurons and other excitable cells, and rapid inactivation of sodium channels is critical for electrical signaling. Scorpion toxins acting on sodium channels and affect the rapid deactivation rate of the channels. Therefore, it is of great importance to analyze the interaction mechanism between scorpion toxins and sodium channels. By comparing the amino acid sequences of Blattella germanica, Periplaneta americana and mammalian sodium channels, it was found that four different amino acids found are I291L, Y292F, V295N, and L406I Site-directed mutation technique was used to construct four amino acid mutant plasmids on the B. germanica VGSCs, and Xenopus oocytes were employed as vehicles for gene expression. Electrophysiological results showed that the V295N mutation significantly reduced the sensitivity of the sodium channel to α-scorpion toxin LqhIT (approximately 21.63-fold). Further 3D homology modeling and molecular docking of the scorpion toxin revealed that the V295N mutation discovered sodium channel M291 to form a hydrogen bond with the K63 amino acid of LqhαIT, which prevented itsbinding with LqhαIT. In conclusion V295N mutation indirectly affected the binding of the channel to LqhαIT, resulting in a reduction in the sensitivity of the channel to LqhαIT. |
| Key words:Blattella germanica sodium channel LqhαIT site-directed mutagenesis two-electrode voltage clamp homologous modeling |
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