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Updated: Aug 12, 2025

Monitoring Protein Adsorption with Solid-state Nanopores

Monitoring Protein Adsorption with Solid-state Nanopores

Published on: December 2, 2011

Nanopore single-molecule biosensor in protein denaturation analysis.

Hong Sun¹, Chuan Yao¹, Kaibo You¹

¹Key Laboratory of Micro-Nano Materials for Energy Storage and Conversion of Henan Province, Institute of Surface Micro and Nano Materials, College of Chemical and Materials Engineering, Xuchang University, Henan, 461000, PR China.

Analytica Chimica Acta

|January 25, 2023

Summary

This summary is machine-generated.

This study introduces a nanopore biosensor for monitoring protein unfolding dynamics. It experimentally confirms a three-state model for lysozyme denaturation, revealing a stable intermediate.

Keywords:

Denaturation mechanism Folding intermediate Lysozyme Nanopore Protein denaturation

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Area of Science:

Biophysics
Protein Chemistry
Nanotechnology

Background:

Protein stability and denaturation mechanisms remain areas of active research.
Understanding these processes is crucial for various biological and biotechnological applications.

Purpose of the Study:

To develop and apply a nanopore single-molecule biosensor for dynamic monitoring of protein folding and unfolding.
To investigate the denaturation mechanism of lysozyme using this novel approach.

Main Methods:

Utilized a nanopore single-molecule biosensor to record ionic current changes.
Gradually increased the concentration of guanidine hydrochloride (GdmCl) to induce protein unfolding.
Analyzed the resulting signal pulses to infer protein conformational changes.

Main Results:

Observed sensitive responses in lysozyme unfolding with increasing GdmCl concentration.
Detected a biphasic pulse, indicating the presence of a stable intermediate.
Provided the first single-molecule experimental confirmation of dynamic equilibrium between native and intermediate states in lysozyme.
Characterized the intermediate as having a globular-like structure, larger gyration radius, and enhanced positive charge density.

Conclusions:

Consolidated the three-state model for lysozyme denaturation.
Supported a 'direct' denaturation model involving electrostatic and van der Waals forces.
Demonstrated the nanopore biosensor's high sensitivity (280 pM detection limit) and reproducibility for analyzing protein folding intermediates.