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Nanopore Detection Using Supercharged Polypeptide Molecular Carriers.

Xiaoyi Wang1, Tina-Marie Thomas2,3, Ren Ren1,4

  • 1Department of Chemistry, Imperial College London, Molecular Science Research Hub, London W12 0BZ, U.K.

Journal of the American Chemical Society
|March 10, 2023
PubMed
Summary
This summary is machine-generated.

Supercharged unstructured polypeptides (SUPs) genetically fused to proteins enable controlled nanopore sensing. This method enhances single-molecule protein detection and differentiation, overcoming current limitations in biological analysis.

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Analytical Chemistry

Background:

  • Single-molecule analysis of proteins is crucial for understanding biological processes and diseases, especially for low-abundance targets.
  • Nanopore sensing offers label-free detection of single proteins, with applications in biomarker screening and drug discovery.
  • Current nanopore sensing faces challenges in controlling protein translocation and correlating structure/function with readouts.

Purpose of the Study:

  • To develop a method for enhanced control and detection of proteins using nanopore sensing.
  • To investigate the use of genetically fused supercharged unstructured polypeptides (SUPs) as molecular carriers for proteins.
  • To overcome spatiotemporal limitations in current single-molecule protein analysis.

Main Methods:

  • Genetically fusing supercharged unstructured polypeptides (SUPs) with target proteins.
  • Utilizing cationic SUPs to modulate protein translocation through nanopores via electrostatic interactions.
  • Analyzing nanopore current signals to differentiate proteins based on size and shape.

Main Results:

  • SUPs significantly slow down protein translocation through nanopores.
  • Characteristic subpeaks in nanopore current allow differentiation of individual proteins.
  • Demonstrated a viable method for controlling molecular transport at the single-molecule level.

Conclusions:

  • Genetically engineered SUPs serve as effective molecular carriers for enhanced nanopore detection of proteins.
  • This approach facilitates single-molecule protein differentiation and interaction studies.
  • SUPs offer a promising strategy to overcome limitations in nanopore sensing for biological applications.