High-throughput chemical proteomics workflow for profiling protein citrullination dynamics.

ABSTRACT: Citrullination is a post-translational modification implicated in autoimmune and inflammatory diseases, yet its low abundance and lack of effective enrichment tools have limited proteome-wide analysis. Here, we develop a robust chemical proteomics workflow with improved specificity and throughput. This method builds upon glyoxal-based derivatization and incorporates a cleavable biotin linker for efficient peptide enrichment, release, and identification via mass spectrometry. Benchmarking demonstrates a > 10-fold increase in the detection of citrullinated peptides at sub-0.1% abundance. Applying this workflow to primary human neutrophils, we successfully monitor dynamic regulation, quantifying dose-dependent activation and inhibition by the PAD4 inhibitor GSK484. Furthermore, stimulation wi

SUBMITTER: Meelker Gonzalez R 

PROVIDER: S-EPMC12932703 | biostudies-literature | 2026 Feb

REPOSITORIES: biostudies-literature