Characterization of the Global Acetylome in Mouse Livers Under a High‐Fat Diet

ABSTRACT Obesity is often linked to protein acetylation and induces intricate changes in post‐translational modifications, affecting mitochondrial regulatory instability. Redox signaling is crucial for maintaining cellular homeostasis. The mechanism by which lysine acetylome drives redox signaling may elucidate the complexity of the biological networks underlying obesity. High‐fat diet (HFD)–fed mouse livers were lysed, immunoprecipitated with lysine‐acetyl antibody‐conjugated beads, and labeled with isotopic 18 O/ 16 O for multiplexing. A total of 2282 proteins and 1384 Kac sites were identified, including 456 newly identified sites that were not matched in the dbPTM and iPTMnet databases. The expression levels of the acetylome were normalized to protein expression, and one‐third of the acetylome was quantified. Differentially expressed acetylated proteins were annotated using Gene Ontology; the upregulated proteins belonged to the peroxisomal pathway. The three proteins involved in redox regulation, glutathione S ‐transferase theta‐1, superoxide dismutase 1, and epoxide hydrolase 1, exhibited significant changes at specific sites with acetyl‐lysine levels in HFD‐fed mouse liver. Our data showed that investigating acetylation pathways can provide insights into the molecular mechanisms driving obesity and offer potential targets for therapeutic interventions.