Proteomic Analysis of Ubiquitination Modified Proteins in Brain Tissue of Rats at Different Times after Severe...

Severe traumatic brain injury (sTBI) is a major global health issue associated with significant morbidity and socioeconomic burden. Although proteomic approaches have identified potential sTBI biomarkers, systematic insights into common pathological mechanisms across different post‑injury phases remain limited. Recent studies indicate altered ubiquitin expression following TBI, yet the specific ubiquitinated proteins involved are not well characterized. Using label‑free ubiquitin proteomics on pooled brain tissue samples from a rat-controlled cortical impact model, we identified 42 differentially ubiquitinated proteins and 29 ubiquitination sites that were commonly observed across 24 h, 72 h, and 7 d post‑injury (based on fold‑change thresholds). Bioinformatics analysis revealed enrichment of these proteins in pathways related to synaptic function, calcium signaling, and cytoskeletal regulation. Parallel reaction monitoring (PRM) confirmed the presence and relative abundance trends of two ubiquitination sites on glial fibrillary acidic protein (GFAP). This descriptive dataset provides a time‑dependent ubiquitination profile after sTBI and highlights candidate proteins for future mechanistic exploration.