iProXProteomicsChu WangMus MusculusBos Taurushttp://www.iprox.org/page/project.html?id=IPX0016779000chuwang@pku.edu.cnPeking UniversityiProXfalseSite-specific Profiling of Lysine Itaconylation via a Thiol-based ProbeLysine itaconylation is a novel post-translational modification (PTM) that was recently discovered in macrophage proteomes mediated by the immunoregulatory metabolite, itaconate. However, there is currently an absence of comprehensive and high-accuracy analytical methods for the global identification of itaconylation sites in proteomes, which limits its biological function study. Here, we developed a thiol-based bioorthogonal probe, BMAyne probe, to site-specifically profile itaconylation in macrophage proteomes by an Activity-based Protein Profiling (ABPP) strategy. Notably, 31 endogenous itaconylation sites on 29 proteins were identified in lipopolysaccharide (LPS)-stimulated macrophages using a thiol-based probe, BMAyne probe, which complemented the results obtained by previous promiscuous antibody enrichment. Our effort provides a unique chemical tool to enrich endogenous lysine itaconylation and establishes a rich database for guiding subsequent functional studies of this unique lysine itaconylation PTM.Tue Apr 21 00:00:00 GMT+01:00 2026PXD077428100909913