An engineered ClyA nanopore detects folded target proteins by selective external association and pore entry.
An engineered ClyA nanopore detects folded target proteins by selective external association and pore entry.
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DOI：
10.1021/nl3024438
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发表时间：
2012-09-12
期刊：
影响因子：
10.8
通讯作者：
Maglia G
中科院分区：
文献类型：
--
作者：
Soskine M;Biesemans A;Moeyaert B;Cheley S;Bayley H;Maglia G
纳米孔已被用于无标记的单分子研究，包括化学反应、核酸分析和在传感方面的应用。作为传感器，生物纳米孔通常比人造纳米孔表现得更好，但它们有一些缺点，包括直径固定。在这里，我们介绍了一种生物纳米孔ClyA，它足够宽，可以采样和区分进入孔腔的大样本蛋白质。值得注意的是，尽管人和牛的凝血酶有86%的序列相同，但仍能引起特征性的离子电流阻断，在−50 mV时，它们的主电流水平相差26±1pA。DNA适体或水飞蓟素作为配基的使用进一步区分了蛋白质分析物。最后，我们构建了带有共价连接适配子的ClyA纳米孔。这些纳米孔选择性地捕获并内化同源蛋白分析物，但排除非同源蛋白分析物，这一过程类似于核孔的运输。
Nanopores have been used in label-free single-molecule studies, including investigations of chemical reactions, nucleic acid analysis and applications in sensing. Biological nanopores generally perform better than artificial nanopores as sensors, but they have disadvantages including a fixed diameter. Here we introduce a biological nanopore ClyA that is wide enough to sample and distinguish large analytes proteins, which enter the pore lumen. Remarkably, human and bovine thrombins, despite 86% sequence identity, elicit characteristic ionic current blockades, which at −50 mV differ in their main current levels by 26 ± 1 pA. The use of DNA aptamers or hirudin as ligands further distinguished the protein analytes. Finally, we constructed ClyA nanopores decorated with covalently attached aptamers. These nanopores selectively captured and internalized cognate protein analytes, but excluded non-cognate analytes, in a process that resembles transport by nuclear pores.
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影响因子：
--
作者：
Maglia, Giovanni;Heron, Andrew J.;Stoddart, David;Japrung, Deanpen;Bayley, Hagan
通讯作者：
Bayley, Hagan
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
影响因子：
46.9
作者：
Movileanu, L;Howorka, S;Bayley, H
通讯作者：
Bayley, H
影响因子：
64.8
作者：
Bayley, H;Cremer, PS
通讯作者：
Cremer, PS
影响因子：
10.8
作者：
Firnkes, Matthias;Pedone, Daniel;Rant, Ulrich
通讯作者：
Rant, Ulrich