Real-time shape approximation and fingerprinting of single proteins using a nanopore

Area of Science:

• Biophysics
• Analytical Chemistry
• Structural Biology

Background:

• Traditional protein characterization methods often require sample modification or cannot analyze individual molecules in solution.
• Ionic current measurements in nanopores offer a way to study single native proteins but have limitations.

Purpose of the Study:

• To develop a novel method for comprehensive single-protein characterization in solution.
• To determine multiple protein properties simultaneously using nanopore technology.

Main Methods:

• Utilizing bilayer-coated solid-state nanopores with zeptolitre sensing volumes.
• Developing a theoretical framework to interpret ionic current modulations caused by protein dynamics.
• Measuring ionic current changes as individual proteins translocate through the nanopore.

Main Results:

• Successfully determined approximate shape, volume, charge, rotational diffusion coefficient, and dipole moment of individual proteins.
• Demonstrated simultaneous measurement of these five key protein parameters.
• Validated the method's potential for identifying and quantifying proteins and protein complexes.

Conclusions:

• The developed nanopore approach enables detailed characterization of single native proteins.
• This method has broad implications for structural biology, proteomics, and biomarker detection.
• Offers a powerful tool for routine protein analysis with potential for high throughput.