Nanopore-Based Detection of Protein Posttranslational Modifications

Area of Science:

• Biochemistry
• Molecular Biology
• Analytical Chemistry

Background:

• Protein posttranslational modifications (PTMs) are vital for cellular functions.
• Conventional methods like mass spectrometry (MS) are sensitive but complex.
• Nanopore sequencing offers label-free, high-throughput analysis for single molecules.

Purpose of the Study:

• To review the current state of nanopore-based PTM detection.
• To outline challenges in applying nanopore technology to protein PTMs.
• To present feasible near-term experimental strategies for protein PTM analysis using nanopores.

Main Methods:

• Review of existing literature on nanopore PTM detection.
• Description of two experimental strategies for protein capture and analysis.
• Focus on label-free, single-molecule detection principles.

Main Results:

• Nanopore technology shows potential for PTM detection, but faces hurdles.
• Challenges include protein handling, unfolding, and PTM diversity.
• Two promising workflows are proposed for practical application.

Conclusions:

• Nanopore sequencing is an emerging tool for protein PTM analysis.
• Overcoming technical challenges is key to widespread adoption.
• The proposed strategies offer a path toward practical nanopore-based PTM identification.