Jingzhaotoxin-X, a gating modifier of Kv4.2 and Kv4.3 potassium channels purified from the venom of the Chinese tarantula Chilobrachys jingzhao
Jingzhaotoxin-X, a gating modifier of Kv4.2 and Kv4.3 potassium channels purified from the venom of the Chinese tarantula Chilobrachys jingzhao
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Jingzhaotoxin-X，Kv4.2 和 Kv4.3 钾通道的门控修饰剂，从中国狼蛛 Chilobrachys jingzhao 的毒液中纯化
DOI：
10.1590/1678-9199-jvatitd-2019-0043
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发表时间：
2020-05
影响因子：
2.4
通讯作者：
Liang Songping
中科院分区：
文献类型：
--
作者：
Deng Meichun;Jiang Liping;Luo Xuan;Tao Huai;Liang Songping
摘要背景：狼蛛是中国境内体型最大的毒蜘蛛之一。在以往的研究中，我们至少从京招蛇毒中提纯并鉴定了8个多肽。在本报告中，我们描述了选择性阻断Kv4.2和Kv4.3钾通道的京招毒素-X(JZTX-X)的纯化和性质。方法：采用阳离子交换高效液相色谱法和反相高效液相色谱法相结合的方法纯化JZTX-X。用自动Edman降解法测定氨基酸序列，用质谱仪(MS)进行确证。在瞬时转染各种离子通道结构的HEK293T细胞中记录到电压门控离子通道电流。此外，还观察了解痛通-X的痛敏作用和S毒素对小鼠运动功能的影响。结果：JZTX-X含有31个氨基酸，其中6个半胱氨酸残基在抑制性半胱氨酸结(ICK)拓扑结构中形成3个二硫键。在全细胞电压钳实验中，JZTX-X以浓度和电压依赖的方式抑制Kv4.2和Kv4.3钾通道，而不影响其他离子通道(Kv1.1，1.2，1.3，2.1，延迟整流钾通道，高和低电压激活的钙通道，以及电压门控钠通道Nav1.5和1.7)。JZTX-X还使电压依赖的通道激活移向更多的去极化电位，而极端的去极化导致可逆的毒素与Kv4.2通道结合。JZTX-X使Kv4.2和Kv4.3的活动转向静息状态，因为在静息电位下，毒素完全抑制通道，即使在没有施加物理刺激的情况下也是如此。鞘内或足底注射JZTX-X可导致机械痛觉阈值(痛觉过敏)的长期下降，但对旋转棒测试中评估的运动功能没有影响。结论：JZTX-X以浓度和电压依赖的方式选择性抑制Kv4.2和Kv4.3钾通道活动，并引起持久的机械性痛敏。
Abstract Background: The tarantula Chilobrachys jingzhao is one of the largest venomous spiders in China. In previous studies, we purified and characterized at least eight peptides from C. jingzhao venom. In this report, we describe the purification and characterization of Jingzhaotoxin-X (JZTX-X), which selectively blocks Kv4.2 and Kv4.3 potassium channels. Methods: JZTX-X was purified using a combination of cation-exchange HPLC and reverse-phase HPLC. The amino-acid sequence was determined by automated Edman degradation and confirmed by mass spectrometry (MS). Voltage-gated ion channel currents were recorded in HEK293t cells transiently transfected with a variety of ion channel constructs. In addition, the hyperalgesic activity of JZTX-X and the toxin´s effect on motor function were assessed in mice. Results: JZTX-X contained 31 amino acids, with six cysteine residues that formed three disulfide bonds within an inhibitory cysteine knot (ICK) topology. In whole-cell voltage-clamp experiments, JZTX-X inhibited Kv4.2 and Kv4.3 potassium channels in a concentration- and voltage-dependent manner, without affecting other ion channels (Kv1.1, 1.2, 1.3, 2.1, delayed rectifier potassium channels, high- and low-voltage-activated Ca2+ channels, and voltage-gated sodium channels Nav1.5 and 1.7). JZTX-X also shifted the voltage-dependent channel activation to more depolarized potentials, whereas extreme depolarization caused reversible toxin binding to Kv4.2 channels. JZTX-X shifted the Kv4.2 and Kv4.3 activities towards a resting state, since at the resting potential the toxin completely inhibited the channels, even in the absence of an applied physical stimulus. Intrathecal or intraplantar injection of JZTX-X caused a long-lasting decrease in the mechanical nociceptive threshold (hyperalgesia) but had no effect on motor function as assessed in the rotarod test. Conclusions: JZTX-X selectively suppresses Kv4.2 and Kv4.3 potassium channel activity in a concentration- and voltage-dependent manner and causes long-lasting mechanical hyperalgesia.
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影响因子：
2.9
作者：
Huai Tao;Jin J Chen;Yueyue Xiao;Yuanyuan Wu;Haibo Su;Dan Li;Heng-Yen Wang;M. Deng;Meixia Wang;Zhong-hua Liu;S. Liang
通讯作者：
Huai Tao;Jin J Chen;Yueyue Xiao;Yuanyuan Wu;Haibo Su;Dan Li;Heng-Yen Wang;M. Deng;Meixia Wang;Zhong-hua Liu;S. Liang
DOI：
10.2210/pdb1emx/pdb
发表时间：
2001-01
期刊：
Protein science : a publication of the Protein Society
影响因子：
--
作者：
Cédric Bernard;Christian Legros;G. Ferrat;Ulrike Bischoff;Annette Marquardt;Olaf Pongs;Hervé Darbon
通讯作者：
Cédric Bernard;Christian Legros;G. Ferrat;Ulrike Bischoff;Annette Marquardt;Olaf Pongs;Hervé Darbon
DOI：
10.1016/j.toxicon.2011.11.021
发表时间：
2012-02
期刊：
Toxicon : official journal of the International Society on Toxinology
影响因子：
--
作者：
C. Yuan;Zhonghua Liu;Weijun Hu;T. Gao;S. Liang
通讯作者：
C. Yuan;Zhonghua Liu;Weijun Hu;T. Gao;S. Liang
DOI：
10.1016/j.toxicon.2008.08.018
发表时间：
2008-12
期刊：
Toxicon : official journal of the International Society on Toxinology
影响因子：
--
作者：
Meichi Wang;J. Diao;Jiang Li;J. Tang;Ying Lin;Weijun Hu;Yongqun Zhang;Yucheng Xiao;S. Liang
通讯作者：
Meichi Wang;J. Diao;Jiang Li;J. Tang;Ying Lin;Weijun Hu;Yongqun Zhang;Yucheng Xiao;S. Liang
影响因子：
5.5
作者：
Sun, Jie;Chen, Shao-Rui;Pan, Hui-Lin
通讯作者：
Pan, Hui-Lin