以水为探针,剖析费摩尔浓度下气溶胶素孔的膜结合机制

IF 9.6 1区 材料科学 Q1 CHEMISTRY, MULTIDISCIPLINARY Nano Letters Pub Date : 2024-11-06 Epub Date: 2024-10-29 DOI:10.1021/acs.nanolett.4c00035
Tereza Roesel, Chan Cao, Juan F Bada Juarez, Matteo Dal Peraro, Sylvie Roke
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引用次数: 0

摘要

气溶胶毒素是一种细菌毒素,可在宿主质膜上形成跨膜孔,具有内径窄、稳定性强等特点。这些优点使其成为检测核酸和肽等生物聚合物的极具潜力的纳米孔。尽管从微生物学和结构的角度来看,人们对气溶胶素已经有了很多了解,但对它的膜关联和孔形成机制还不完全清楚。在这里,我们利用角度分辨二次谐波散射(AR-SHS)和单通道电流测量来研究野生型(wt)气溶蛋白及其突变体如何在飞摩尔浓度的水溶液中与脂质体相互作用。我们的AR-SHS实验非常灵敏,足以检测到膜结合气溶胶素静电特性的变化,这些变化是由pH值变化引起的。我们首次报道了气溶胶素在孔形成机制不同阶段的膜结合亲和力:wt气溶胶素的结合亲和力高达20 fM,而准孔和预孔态的膜结合亲和力逐渐降低,插入不完全,并出现孔开放特征。此外,我们还定量分析了与纳米孔传感应用相关的突变体的膜亲和性。我们的研究为有效筛选生物孔提供了一种无标记方法,这种方法适用于分子传感和测序测量以及孔形成过程的探测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Dissecting the Membrane Association Mechanism of Aerolysin Pores at Femtomolar Concentrations Using Water as a Probe.

Aerolysin is a bacterial toxin that forms transmembrane pores at the host plasma membrane and has a narrow internal diameter and great stability. These assets make it a highly promising nanopore for detecting biopolymers such as nucleic acids and peptides. Although much is known about aerolysin from a microbiological and structural perspective, its membrane association and pore-formation mechanism are not yet fully understood. Here, we used angle-resolved second harmonic scattering (AR-SHS) and single-channel current measurements to investigate how wild-type (wt) aerolysin and its mutants interact with liposomes in aqueous solutions at femtomolar concentrations. Our AR-SHS experiments were sensitive enough to detect changes in the electrostatic properties of membrane-bound aerolysin, which were induced by variations in pH levels. We reported for the first time the membrane binding affinity of aerolysin at different stages of the pore formation mechanism: while wt aerolysin has a binding affinity as high as 20 fM, the quasi-pore and the prepore states show gradually decreasing membrane affinities, incomplete insertion, and a pore opening signature. Moreover, we quantitatively characterized the membrane affinity of mutants relevant for applications to nanopore sensing. Our study provides a label-free method for efficiently screening biological pores suitable for conducting molecular sensing and sequencing measurements as well as for probing pore-forming processes.

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来源期刊
Nano Letters
Nano Letters 工程技术-材料科学:综合
CiteScore
16.80
自引率
2.80%
发文量
1182
审稿时长
1.4 months
期刊介绍: Nano Letters serves as a dynamic platform for promptly disseminating original results in fundamental, applied, and emerging research across all facets of nanoscience and nanotechnology. A pivotal criterion for inclusion within Nano Letters is the convergence of at least two different areas or disciplines, ensuring a rich interdisciplinary scope. The journal is dedicated to fostering exploration in diverse areas, including: - Experimental and theoretical findings on physical, chemical, and biological phenomena at the nanoscale - Synthesis, characterization, and processing of organic, inorganic, polymer, and hybrid nanomaterials through physical, chemical, and biological methodologies - Modeling and simulation of synthetic, assembly, and interaction processes - Realization of integrated nanostructures and nano-engineered devices exhibiting advanced performance - Applications of nanoscale materials in living and environmental systems Nano Letters is committed to advancing and showcasing groundbreaking research that intersects various domains, fostering innovation and collaboration in the ever-evolving field of nanoscience and nanotechnology.
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