Screening for voltage-gated sodium channel interacting peptides.

Screening for voltage-gated sodium channel interacting peptides.
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筛选电压门控钠通道相互作用肽。

DOI:
10.1038/srep04569
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发表时间:
2014-04-02
期刊:
影响因子:
4.6
通讯作者:
Zhang DY
Zhang DY
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Meng E;Cai TF;Zhang H;Tang S;Li MJ;Li WY;Huang PF;Liu K;Wu L;Zhu LY;Liu L;Peng K;Dai XD;Jiang H;Zeng XZ;Liang SP;Zhang DY

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电压门控钠通道(VGSC)相互作用肽在基础研究和药物研究中都具有特殊的意义。在这项研究中,我们建立了一种基于酵母-双杂交的策略来检测神经毒肽与VGSC细胞外区域之间的相互作用。使用先前报道的神经毒素JZTX-III作为模型分子,我们证明了JZTX-III与其靶细胞外区域hNav1.5之间的相互作用是可检测的,并且检测到的相互作用与其活性直接相关。我们进一步将这一策略应用于VGSC相互作用肽的筛选。以hNav1.5细胞外区为诱饵,从随机肽库中鉴定出一种新的钠通道抑制剂SSCM-1。该肽在全细胞膜片钳实验中选择性抑制hNav1.5电流。该策略可用于VGSCs或其他离子通道的靶向性相互作用肽的大规模筛选。
The voltage-gated sodium channel (VGSC) interacting peptide is of special interest for both basic research and pharmaceutical purposes. In this study, we established a yeast-two-hybrid based strategy to detect the interaction(s) between neurotoxic peptide and the extracellular region of VGSC. Using a previously reported neurotoxin JZTX-III as a model molecule, we demonstrated that the interactions between JZTX-III and the extracellular regions of its target hNav1.5 are detectable and the detected interactions are directly related to its activity. We further applied this strategy to the screening of VGSC interacting peptides. Using the extracellular region of hNav1.5 as the bait, we identified a novel sodium channel inhibitor SSCM-1 from a random peptide library. This peptide selectively inhibits hNav1.5 currents in the whole-cell patch clamp assays. This strategy might be used for the large scale screening for target-specific interacting peptides of VGSCs or other ion channels.
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