Ribosome Fingerprinting with a Solid-State Nanopore

ACS Sens. 2020 Nov 25;5(11):3533-3539. doi: 10.1021/acssensors.0c01642. Epub 2020 Oct 28.

Abstract

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.

Keywords: nanopipette; nanopore; polysome; ribosome; single-molecule; translation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Humans
  • Nanopores*
  • Polyribosomes / metabolism
  • Protein Biosynthesis
  • RNA, Messenger / metabolism
  • Ribosomes / metabolism

Substances

  • RNA, Messenger