Purification and Characterization of JZTx-14, a Potent Antagonist of Mammalian and Prokaryotic Voltage-Gated Sodium Channels.
Purification and Characterization of JZTx-14, a Potent Antagonist of Mammalian and Prokaryotic Voltage-Gated Sodium Channels.
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DOI：
10.3390/toxins10100408
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发表时间：
2018-10-10
期刊：
影响因子：
4.2
通讯作者：
Liu Z
中科院分区：
文献类型：
--
作者：
Zhang J;Tang D;Liu S;Hu H;Liang S;Tang C;Liu Z
探索配体与电压门控钠通道（NaVs）的相互作用促进了我们对其药理学的理解。在此，我们报告了一种新的非选择性的哺乳动物和细菌的NaVs毒素，JZTx-14，从蜘蛛Chilobrachys jingzhao的毒液的纯化和表征。该毒素对哺乳动物NaV1.2-1.8通道和细菌NaChBac通道的峰电流具有较低的IC 50值（<1 µM），主要抑制NaV1.9通道的快速失活。对NaV1.5/NaV1.9嵌合通道的分析表明，NaV1.5结构域II S3-4环参与毒素缔合。从研究毒素-NaV 1.2通道相互作用获得的动力学数据显示，JZTx-14是一种门控修饰剂，可能将通道捕获在静息状态;然而，它与位点4毒素HNTx-III的不同之处在于不可逆地阻断NaV电流并显示与通道的状态无关性结合。JZTx-14可能稳定地结合到NaV1.2-1.8结构域II电压传感器深处的保守毒素口袋，而不管通道构象变化如何，并且其对NaV的作用需要毒素将S3-4环捕获在其静息状态。对于NaChBac通道，JZTx-14正移其电导-电压（G-V）和稳态失活关系。NaChBac S3-4环的丙氨酸扫描分析揭示，第108位苯丙氨酸（F108）是决定JZTx-14-NaChBac相互作用的关键残基。综上所述，本研究为JZTx-14提供了对祖先细菌NaVs和高度进化的后代哺乳动物NaVs的有效但混杂的抑制活性，并且它是了解NaVs药理学的有用探针。
Exploring the interaction of ligands with voltage-gated sodium channels (NaVs) has advanced our understanding of their pharmacology. Herein, we report the purification and characterization of a novel non-selective mammalian and bacterial NaVs toxin, JZTx-14, from the venom of the spider Chilobrachys jingzhao. This toxin potently inhibited the peak currents of mammalian NaV1.2–1.8 channels and the bacterial NaChBac channel with low IC50 values (<1 µM), and it mainly inhibited the fast inactivation of the NaV1.9 channel. Analysis of NaV1.5/NaV1.9 chimeric channel showed that the NaV1.5 domain II S3–4 loop is involved in toxin association. Kinetics data obtained from studying toxin–NaV1.2 channel interaction showed that JZTx-14 was a gating modifier that possibly trapped the channel in resting state; however, it differed from site 4 toxin HNTx-III by irreversibly blocking NaV currents and showing state-independent binding with the channel. JZTx-14 might stably bind to a conserved toxin pocket deep within the NaV1.2–1.8 domain II voltage sensor regardless of channel conformation change, and its effect on NaVs requires the toxin to trap the S3–4 loop in its resting state. For the NaChBac channel, JZTx-14 positively shifted its conductance-voltage (G–V) and steady-state inactivation relationships. An alanine scan analysis of the NaChBac S3–4 loop revealed that the 108th phenylalanine (F108) was the key residue determining the JZTx-14–NaChBac interaction. In summary, this study provided JZTx-14 with potent but promiscuous inhibitory activity on both the ancestor bacterial NaVs and the highly evolved descendant mammalian NaVs, and it is a useful probe to understand the pharmacology of NaVs.
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影响因子：
64.8
作者：
Payandeh, Jian;El-Din, Tamer M. Gamal;Scheuer, Todd;Zheng, Ning;Catterall, William A.
通讯作者：
Catterall, William A.
影响因子：
64.8
作者：
Jiang D;Gamal El-Din TM;Ing C;Lu P;Pomès R;Zheng N;Catterall WA
通讯作者：
Catterall WA
DOI：
10.1085/jgp.201210779
发表时间：
2012-06
期刊：
The Journal of general physiology
影响因子：
--
作者：
Lee S;Goodchild SJ;Ahern CA
通讯作者：
Ahern CA
影响因子：
3.4
作者：
Pavlov, E;Bladen, C;French, RJ
通讯作者：
French, RJ
影响因子：
4.2
作者：
Green BR;Bulaj G;Norton RS
通讯作者：
Norton RS