Site-specific post-translational modification detection by polar charged engineered MspA nanopores

nanopore sensing-lysine crotonylation and succinylation. Furthermore, the nanopore is capable of distinguishing subtle positional isomers. Coupled with a machine learning algorithm that classifies single-molecule events with >98% accuracy, our findings establish a generalizable principle of electrostatic gating for PTM profiling, offering a versatile chemical tool for next-generation single-molecule proteomics.