Electrical recognition of the twenty proteinogenic amino acids using an aerolysin nanopore.,Nature Biotechnology

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Electrical recognition of the twenty proteinogenic amino acids using an aerolysin nanopore.
Nature Biotechnology ( IF 33.1 ) Pub Date : 2019-12-16 , DOI: 10.1038/s41587-019-0345-2
Hadjer Ouldali ₁ , Kumar Sarthak ₂ , Tobias Ensslen ₃ , Fabien Piguet _{1,

4} , Philippe Manivet _{5,

6} , Juan Pelta ₇ , Jan C Behrends _{3,

8,

9} , Aleksei Aksimentiev _{2,

10,

11} , Abdelghani Oukhaled ₁

Affiliation

LAMBE UMR 8587, Université de Cergy-Pontoise, CNRS, CEA, Université Paris-Seine, Cergy-Pontoise, France.
Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Laboratory for Membrane Physiology and Technology, Department of Physiology, Faculty of Medicine, University of Freiburg, Freiburg, Germany.
DreamPore S.A.S., 33 Boulevard du Port 95000, Cergy, France.
APHP, GHU APHP.Nord, DMU BioGem, Hôpital Lariboisière, BIOBANK Lariboisière Department BB-0033-00064, Plateforme de BioPathologie et de Technologies Innovantes en Santé, Paris, France.
INSERM UMR 1141 "NeuroDiderot", Université de Paris, Paris, France.
LAMBE UMR 8587, Université d'Evry-Val-d'Essonne, CNRS, CEA, Université Paris-Saclay, Evry, France.
Freiburg Materials Research Centre (FMF), University of Freiburg, Freiburg, Germany.
Freiburg Centre for Interactive Materials and Bioinspired Technologies (FIT), University of Freiburg, Freiburg, Germany.
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, USA.
Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL, USA.

Efforts to sequence single protein molecules in nanopores1-5 have been hampered by the lack of techniques with sufficient sensitivity to discern the subtle molecular differences among all twenty amino acids. Here we report ionic current detection of all twenty proteinogenic amino acids in an aerolysin nanopore with the help of a short polycationic carrier. Application of molecular dynamics simulations revealed that the aerolysin nanopore has a built-in single-molecule trap that fully confines a polycationic carrier-bound amino acid inside the sensing region of the aerolysin. This structural feature means that each amino acid spends sufficient time in the pore for sensitive measurement of the excluded volume of the amino acid. We show that distinct current blockades in wild-type aerolysin can be used to identify 13 of the 20 natural amino acids. Furthermore, we show that chemical modifications, instrumentation advances and nanopore engineering offer a route toward identification of the remaining seven amino acids. These findings may pave the way to nanopore protein sequencing.

中文翻译：

使用气溶素纳米孔对二十种蛋白质氨基酸进行电识别。

由于缺乏足够灵敏的技术来辨别所有二十种氨基酸之间细微的分子差异，对纳米孔 1-5 中的单个蛋白质分子进行测序的努力受到了阻碍。在这里，我们报告了借助短聚阳离子载体对气溶素纳米孔中所有二十种蛋白氨基酸的离子电流检测。分子动力学模拟的应用表明，气溶素纳米孔具有内置的单分子陷阱，可将聚阳离子载体结合的氨基酸完全限制在气溶素的传感区域内。这种结构特征意味着每种氨基酸在孔中停留足够的时间，以便灵敏地测量氨基酸的排除体积。我们证明，野生型气溶素中不同的电流阻断可用于识别 20 种天然氨基酸中的 13 种。此外，我们表明化学修饰、仪器进步和纳米孔工程为鉴定其余七种氨基酸提供了一条途径。这些发现可能为纳米孔蛋白质测序铺平道路。

更新日期：2019-12-17

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