MassIVE MSV000097617

Partial Public PXD062929

High-throughput chemical proteomics workflow for profiling protein citrullination dynamics

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Description

Citrullination is a post-translational modification implicated in autoimmune and inflammatory diseases but remains challenging to analyze biochemically. Its low stoichiometry and the lack of enrichment tools have hindered its global and site-specific profiling. Here, we developed a high-throughput chemical proteomics workflow for citrullination profiling with enhanced sensitivity and specificity. This workflow employs glyoxal-based derivatization of citrullinated peptides, followed by a click reaction with a biotin conjugate, featuring a cleavable linker that enhances peptide recovery and following identification. We examined the derivatization efficiency using synthetic citrullinated peptides and demonstrated its sensitivity and reproducibility in complex biological samples. Mouse brain digest samples were then used for direct comparison to existent enrichment tools. Dose-dependent ionomycin treatments of isolated neutrophils from fresh blood allowed us to profile citrullination during neutrophil extracellular trap (NET) formation. Known and novel sites on different histone proteins, as well as structural proteins mediating nuclear and cytoskeletal reorganization, were found to be regulated. These findings illuminate citrullination dynamics in NET formation and establish a platform for exploring its roles in diverse systems. [doi:10.25345/C5ZS2KR75] [dataset license: CC0 1.0 Universal (CC0 1.0)]

Keywords: Citrullination ; Enrichment ; Chemical derivatization ; Neutrophil activation ; Mouse brain ; Histone ; DatasetType:Proteomics

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Principal Investigators:
(in alphabetical order) 		Dr. Chien-Yun Lee, Young Investigator Group: Mass Spectrometry in Systems Neurosciences, School of Life Sciences,Technical University of Munich, Germany
Submitting User: Rebecca_Meelker
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Distinct protein accessions are counted across all files submitted in the "Statistical Analysis of Quantified Analytes" category having a "Protein" column in this dataset.

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