Protein S-palmitoylation is a dynamic and reversible post-translational modification (PTM) that governs diverse cellular processes, yet its study remains constrained by the indirect and ensemble-averaging nature of classical biochemical assays. Here, researchers report a nanopore-based platform, PALM-Scan (Palmitoylation Analysis via Label-free Monitoring and Signature Capture on Nanopore) that directly transduces physicochemical signatures of S-palmitoylated peptides into digital electrical signals. Using an engineered Mycobacterium smegmatis porin A nanopore, PALM-Scan enables sequence-independent identification of S-palmitoylation, and simultaneously differentiates S-palmitoylation from other cysteine-directed PTMs, including S-nitrosylation and S-glutathionylation, without probes or enrichment. Researchers further demonstrate its translational potential by successfully detecting and distinguishing S-palmitoylated subpopulations of disease-relevant biomarkers, including beclin-1 (BECN1)- and glial fibrillary acidic protein (GFAP)-derived peptides, in complex mixtures with minimal sample input. In summary, by integrating label-free operation, single-molecule resolution, and multiplex discrimination, PALM-Scan provides a transformative tool for interrogating S-palmitoylation dynamics in both basic research and biomedical applications.

This work entitled “PALM-Scan Enables Single-Molecule Site-Specific Detection of Protein S-Palmitoylation with a Nanopore” is published on《Molecular Cell》, Dr. Liang Wang (Chongqing Institute of Green and Intelligent Technology, Chinese Academy of Sciences) and Dr. Aimin Yang (Chongqing University) are corresponding authors. The work is supported by the National Key Research and Development Program of China, the National Natural Science Foundation of China, Chongqing Talents-Exceptional Young Talents Project, Natural Science Foundation of Chongqing, Youth Innovation Promotion Association of Chinese Academy of Sciences, the Venture Innovation Support Program for Chongqing Overseas Returnees, and the Fundamental Research Funds for the Central Universities.

Figure 1. PALM-Scan platform（Palmitoylation Analysis via Label-free Monitoring and Signature Capture on Nanopore）