Mapping the sensing spots of aerolysin for single oligonucleotides analysis.
Mapping the sensing spots of aerolysin for single oligonucleotides analysis.
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DOI：
10.1038/s41467-018-05108-5
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发表时间：
2018-07-19
影响因子：
16.6
通讯作者：
Long YT
中科院分区：
文献类型：
--
作者：
Cao C;Li MY;Cirauqui N;Wang YQ;Dal Peraro M;Tian H;Long YT
纳米孔传感是一种强大的单分子DNA和蛋白质测序方法。最近的研究表明，气溶素表现出高灵敏度的单分子检测。然而，气溶素孔的原子分辨率结构的缺乏阻碍了对其传感能力的理解。在这里，我们整合纳米孔实验结果和分子模拟的基础上，最近的孔结构模型，精确地映射这种毒素的ssDNA易位的传感点。合理探测ssDNA的长度和组成孔易位提供了新的重要见解气溶素纳米孔的分子决定因素。计算和实验结果揭示了两个关键的传感点（R220，K238）产生两个收缩点沿着孔腔。利用传感点，所有四个核碱基，胞嘧啶甲基化和鸟嘌呤氧化可以清楚地确定在混合物样品。这些结果为气溶素作为DNA测序纳米传感器的潜力提供了证据。纳米孔是一种新兴的强大的单分子DNA测序方法。在这里，作者绘制了气溶素的结构，用作纳米孔，并显示了修饰和未修饰的核碱基的检测。
Nanopore sensing is a powerful single-molecule method for DNA and protein sequencing. Recent studies have demonstrated that aerolysin exhibits a high sensitivity for single-molecule detection. However, the lack of the atomic resolution structure of aerolysin pore has hindered the understanding of its sensing capabilities. Herein, we integrate nanopore experimental results and molecular simulations based on a recent pore structural model to precisely map the sensing spots of this toxin for ssDNA translocation. Rationally probing ssDNA length and composition upon pore translocation provides new important insights for molecular determinants of the aerolysin nanopore. Computational and experimental results reveal two critical sensing spots (R220, K238) generating two constriction points along the pore lumen. Taking advantage of the sensing spots, all four nucleobases, cytosine methylation and oxidation of guanine can be clearly identified in a mixture sample. The results provide evidence for the potential of aerolysin as a nanosensor for DNA sequencing. Nanopores are an emerging powerful single-molecule method of DNA sequencing. Here the authors map the structure of aerolysin for use as a nanopore and show detection of modified and unmodified nucleobases.
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