Macromolecular Crowding Enhances the Detection of DNA and Proteins by a Solid-State Nanopore.,Nano Letters

当前位置： X-MOL 学术 › Nano Lett. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Macromolecular Crowding Enhances the Detection of DNA and Proteins by a Solid-State Nanopore.
Nano Letters ( IF 9.6 ) Pub Date : 2020-06-19 , DOI: 10.1021/acs.nanolett.0c02246
Chalmers C Chau _{1,

2} , Sheena E Radford ₁ , Eric W Hewitt ₁ , Paolo Actis ₂

Affiliation

Nanopore analysis of nucleic acid is now routine, but detection of proteins remains challenging. Here, we report the systematic characterization of the effect of macromolecular crowding on the detection sensitivity of a solid-state nanopore for circular and linearized DNA plasmids, globular proteins (β-galactosidase), and filamentous proteins (α-synuclein amyloid fibrils). We observe a remarkable ca. 1000-fold increase in the molecule count for the globular protein β-galactosidase and a 6-fold increase in peak amplitude for plasmid DNA under crowded conditions. We also demonstrate that macromolecular crowding facilitates the study of the topology of DNA plasmids and the characterization of amyloid fibril preparations with different length distributions. A remarkable feature of this method is its ease of use; it simply requires the addition of a macromolecular crowding agent to the electrolyte. We therefore envision that macromolecular crowding can be applied to many applications in the analysis of biomolecules by solid-state nanopores.

中文翻译：

大分子拥挤增强了固态纳米孔对 DNA 和蛋白质的检测。

核酸的纳米孔分析现已成为常规，但蛋白质的检测仍然具有挑战性。在这里，我们报告了大分子拥挤对固态纳米孔对圆形和线性 DNA 质粒、球状蛋白（β-半乳糖苷酶）和丝状蛋白（α-突触核蛋白淀粉样原纤维）检测灵敏度影响的系统表征。我们观察到一个显着的大约。在拥挤条件下，球状蛋白 β-半乳糖苷酶的分子计数增加了 1000 倍，质粒 DNA 的峰值幅度增加了 6 倍。我们还证明，大分子拥挤有助于研究 DNA 质粒的拓扑结构和不同长度分布的淀粉样原纤维制剂的表征。这种方法的一个显着特点是它的易用性；它只需要在电解液中添加高分子拥挤剂即可。因此，我们设想大分子拥挤可以应用于固态纳米孔分析生物分子的许多应用。

更新日期：2020-07-08

点击分享查看原文

点击收藏

公开下载

阅读更多本刊新发论文本刊介绍/投稿指南