Nanopore sensing of botulinum toxin type B by discriminating an enzymatically cleaved Peptide from a synaptic protein synaptobrevin 2 derivative.
Nanopore sensing of botulinum toxin type B by discriminating an enzymatically cleaved Peptide from a synaptic protein synaptobrevin 2 derivative.
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DOI：
10.1021/am5056596
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发表时间：
2015-01-14
影响因子：
9.5
通讯作者：
Gu, Li-Qun
中科院分区：
文献类型：
--
作者：
Wang, Yong;Montana, Vedrana;Grubisic, Vladimir;Stout, Randy F., Jr.;Parpura, Vladimir;Gu, Li-Qun
肉毒杆菌神经毒素是人类已知的最致命的毒素。生物防御需要及早、快速地发现bont。传统上，bont可以通过在注射了人类物质、血清或粪便的小鼠身上寻找肉毒杆菌中毒的迹象来检测。虽然活体动物试验仍然是最敏感的方法，但它成本高，速度慢，并与法律和伦理限制有关。各种生物化学、光学和机械方法已发展用于bont检测，速度提高，但灵敏度较低。在这里，我们报道了一种新型的基于纳米孔的BoNT B型（BoNT-B）传感器，可以监测毒素在其底物（重组突触蛋白synaptobrevin 2衍生物）上的酶活性。通过分析由特定的BoNT-B消化肽作为标记引起的孔电流调制，在几分钟内确定了亚纳摩尔浓度的BoNT-B的存在。纳米孔探测器将填补神经毒素快速、高灵敏度检测的空白，并为探索生物聚合物运输的生物物理机制提供了一个很好的系统。
Botulinum neurotoxins (BoNTs) are the most lethal toxin known to human. Biodefense requires early and rapid detection of BoNTs. Traditionally, BoNTs can be detected by looking for signs of botulism in mice that receive an injection of human material, serum or stool. While the living animal assay remains the most sensitive approach, it is costly, slow and associated with legal and ethical constrains. Various biochemical, optical and mechanical methods have been developed for BoNTs detection with improved speed, but with lesser sensitivity. Here, we report a novel nanopore-based BoNT type B (BoNT-B) sensor that monitors the toxin’s enzymatic activity on its substrate, a recombinant synaptic protein synaptobrevin 2 derivative. By analyzing the modulation of the pore current caused by the specific BoNT-B-digested peptide as a marker, the presence of BoNT-B at a subnanomolar concentration was identified within minutes. The nanopore detector would fill the niche for a much needed rapid and highly sensitive detection of neurotoxins, and provide an excellent system to explore biophysical mechanisms for biopolymer transportation.
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影响因子：
--
作者：
Bayley H;Cronin B;Heron A;Holden MA;Hwang WL;Syeda R;Thompson J;Wallace M
通讯作者：
Wallace M
影响因子：
14.8
作者：
Degiacomi, Matteo T.;Lacovache, Ioan;Dal Peraro, Matteo
通讯作者：
Dal Peraro, Matteo
DOI：
10.1073/pnas.2531778100
发表时间：
2003-12-23
影响因子：
11.1
作者：
Gu, LQ;Cheley, S;Bayley, H
通讯作者：
Bayley, H
DOI：
10.1085/jgp.118.5.481
发表时间：
2001-11
期刊：
The Journal of general physiology
影响因子：
--
作者：
Gu LQ;Cheley S;Bayley H
通讯作者：
Bayley H
影响因子：
46.9
作者：
通讯作者：
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