Nanopore-Based High-Resolution Detection of Multiple Post-Translational Modifications in Protein

Area of Science:

• Biochemistry
• Nanotechnology
• Analytical Chemistry

Background:

• Protein post-translational modifications (PTMs) are vital for cellular functions.
• Limited availability of reliable and cost-effective sensing methods hinders PTM research at the proteome level.

Purpose of the Study:

• To develop a straightforward and effective strategy for detecting PTMs on short peptides.
• To establish a host-guest interaction-assisted nanopore sensing approach for PTM identification.

Main Methods:

• Utilized α-hemolysin nanopores and cucurbit[7]uril for host-guest interactions.
• Analyzed current blockage during peptide translocation through the nanopore.
• Incorporated a short peptide probe to enhance discrimination capabilities.

Main Results:

• Successfully identified 13 types of PTMs at a specific position in phenylalanine-containing peptides.
• Demonstrated discrimination of various PTMs, positional isomers, and multiple PTMs using the developed strategy.
• Achieved accurate PTM detection via nanopore sensing.

Conclusions:

• The developed host-guest interaction-assisted nanopore sensing strategy offers a promising method for PTM detection.
• This approach provides an efficient alternative to traditional mass spectrometry for analyzing PTMs in biological samples.