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Related Concept Videos

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Sequencing of mRNA from Whole Blood using Nanopore Sequencing
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Ribosome Fingerprinting with a Solid-State Nanopore.

Mukhil Raveendran1, Anna Rose Leach1, Tayah Hopes2,3

  • 1School of Electronic and Electrical Engineering and Pollard Institute, University of Leeds, Leeds LS2 9JT, U.K.

ACS Sensors
|October 28, 2020
PubMed
Summary

Solid-state nanopores can distinguish between individual ribosomes and polysomes. This technique differentiates polysome sizes, offering a rapid method for analyzing these crucial molecular machines.

Keywords:
nanopipettenanoporepolysomeribosomesingle-moleculetranslation

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Area of Science:

  • Molecular Biology
  • Biophysics
  • Nanotechnology

Background:

  • Ribosomes are essential macromolecular machines responsible for protein synthesis via mRNA translation.
  • Analyzing ribosome complexes, such as 80S ribosomes and polysomes, is crucial for understanding cellular function.
  • Current methods for characterizing these complexes can be complex and time-consuming.

Purpose of the Study:

  • To investigate the potential of solid-state nanopores for analyzing and differentiating ribosomal complexes.
  • To characterize 80S ribosomes and polysomes from human and Drosophila melanogaster cells using nanopore technology.
  • To establish whether nanopore measurements can discriminate between different types and sizes of ribosomal complexes.

Main Methods:

  • Utilizing solid-state nanopores to measure the electrical translocation of individual ribosomal complexes.
  • Analyzing the peak amplitude and dwell time characteristics of translocating 80S ribosomes and polysomes.
  • Comparing nanopore data from mixed samples of 80S ribosomes and polysomes.

Main Results:

  • 80S ribosomes and polysomes exhibit distinct peak amplitude and dwell time signatures in nanopore measurements.
  • Solid-state nanopores successfully discriminated between 80S ribosomes and polysomes in mixed samples.
  • The study differentiated between large polysomes (more than seven ribosomes) and smaller polysomes (two to three ribosomes).
  • A correlation was observed between polysome size and measured peak amplitude.

Conclusions:

  • Solid-state nanopore analysis provides a rapid and effective method for detecting and characterizing ribosomal complexes.
  • Nanopore technology offers a promising approach for distinguishing between 80S ribosomes and polysomes based on their unique physical properties.
  • This technique has the potential to advance the study of protein synthesis and translational regulation.