Raveendran Mukhil, Leach Anna Rose, Hopes Tayah, Aspden Julie L, Actis Paolo
School of Electronic and Electrical Engineering and Pollard Institute, University of Leeds, Leeds LS2 9JT, U.K.
School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Leeds LS2 9JT, U.K.
ACS Sens. 2020 Nov 25;5(11):3533-3539. doi: 10.1021/acssensors.0c01642. Epub 2020 Oct 28.
Nanopores hold great potential for the analysis of complex biological molecules at the single-entity level. One particularly interesting macromolecular machine is the ribosome, responsible for translating mRNAs into proteins. In this study, we use a solid-state nanopore to fingerprint 80S ribosomes and polysomes from a human neuronal cell line and cultured cells and ovaries. Specifically, we show that the peak amplitude and dwell time characteristics of 80S ribosomes are distinct from polysomes and can be used to discriminate ribosomes from polysomes in mixed samples. Moreover, we are able to distinguish large polysomes, containing more than seven ribosomes, from those containing two to three ribosomes, and demonstrate a correlation between polysome size and peak amplitude. This study highlights the application of solid-state nanopores as a rapid analytical tool for the detection and characterization of ribosomal complexes.
纳米孔在单实体水平分析复杂生物分子方面具有巨大潜力。一种特别有趣的大分子机器是核糖体,它负责将信使核糖核酸(mRNA)翻译成蛋白质。在本研究中,我们使用固态纳米孔对来自人类神经细胞系、培养细胞和卵巢的80S核糖体及多核糖体进行指纹识别。具体而言,我们表明80S核糖体的峰值幅度和驻留时间特征与多核糖体不同,可用于在混合样本中区分核糖体和多核糖体。此外,我们能够区分含有七个以上核糖体的大多核糖体和含有两到三个核糖体的多核糖体,并证明多核糖体大小与峰值幅度之间存在相关性。这项研究突出了固态纳米孔作为一种快速分析工具在核糖体复合物检测和表征方面的应用。