Ribosome Fingerprinting with a Solid-State Nanopore

被引:24
作者
Raveendran, Mukhil [1 ,2 ]
Leach, Anna Rose [1 ,2 ]
Hopes, Tayah [3 ,4 ]
Aspden, Julie L. [3 ,4 ]
Actis, Paolo [1 ,2 ,4 ,5 ]
机构
[1] Univ Leeds, Sch Elect & Elect Engn, Leeds LS2 9JT, W Yorkshire, England
[2] Univ Leeds, Pollard Inst, Leeds LS2 9JT, W Yorkshire, England
[3] Univ Leeds, Fac Biol Sci, Sch Mol & Cellular Biol, Leeds LS2 9JT, W Yorkshire, England
[4] Univ Leeds, LeedsOm, Leeds LS2 9JT, W Yorkshire, England
[5] Bragg Ctr Mat Res, Leeds LS2 9JT, W Yorkshire, England
基金
英国惠康基金; 英国生物技术与生命科学研究理事会;
关键词
nanopore; nanopipette; ribosome; single-molecule; polysome; translation;
D O I
10.1021/acssensors.0c01642
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
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 andDrosophila melanogaster 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.
引用
收藏
页码:3533 / 3539
页数:7
相关论文
共 41 条
[1]   Chemically Modified Hydrogel-Filled Nanopores: A Tunable Platform for Single-Molecule Sensing [J].
Al Sulaiman, Dana ;
Cadinu, Paolo ;
Ivanov, Aleksandar P. ;
Edel, Joshua B. ;
Ladame, Sylvain .
NANO LETTERS, 2018, 18 (09) :6084-6093
[2]   Single-Molecule Analysis with Solid-State Nanopores [J].
Albrecht, Tim .
ANNUAL REVIEW OF ANALYTICAL CHEMISTRY, VOL 12, 2019, 12 :371-387
[3]   Proteomic analysis of polyribosomes identifies splicing factors as potential regulators of translation during mitosis [J].
Aviner, Ranen ;
Hofmann, Sarah ;
Elman, Tamar ;
Shenoy, Anjana ;
Geiger, Tamar ;
Elkon, Ran ;
Ehrlich, Marcelo ;
Elroy-Stein, Orna .
NUCLEIC ACIDS RESEARCH, 2017, 45 (10) :5945-5957
[4]  
Bakowska-Zywicka K., 2012, COLD SPRING HARB PER, V4
[5]   Resistive-pulse sensing - From microbes to molecules [J].
Bayley, H ;
Martin, CR .
CHEMICAL REVIEWS, 2000, 100 (07) :2575-2594
[6]   Structural Snapshots of Actively Translating Human Ribosomes [J].
Behrmann, Elmar ;
Loerke, Justus ;
Budkevich, Tatyana V. ;
Yamamoto, Kaori ;
Schmidt, Andrea ;
Penczek, Pawel A. ;
Vos, Matthijn R. ;
Buerger, Joerg ;
Mielke, Thorsten ;
Scheerer, Patrick ;
Spahn, Christian M. T. .
CELL, 2015, 161 (04) :845-857
[7]   Genome-wide analysis of transcript abundance and translation in arabidopsis seedlings subjected to oxygen deprivation [J].
Branco-Price, C ;
Kawaguchi, R ;
Ferreira, RB ;
Bailey-Serres, J .
ANNALS OF BOTANY, 2005, 96 (04) :647-660
[8]   HIGH-RESOLUTION DENSITY GRADIENT SEDIMENTATION ANALYSIS [J].
BRITTEN, RJ ;
ROBERTS, RB .
SCIENCE, 1960, 131 (3392) :32-33
[9]   Analysis of translation using polysome profiling [J].
Chasse, Heloise ;
Boulben, Sandrine ;
Costache, Vlad ;
Cormier, Patrick ;
Morales, Julia .
NUCLEIC ACIDS RESEARCH, 2017, 45 (03) :e15
[10]   Macromolecular Crowding Enhances the Detection of DNA and Proteins by a Solid-State Nanopore [J].
Chau, Chalmers C. ;
Radford, Sheena E. ;
Hewitt, Eric W. ;
Actis, Paolo .
NANO LETTERS, 2020, 20 (07) :5553-5561