ABSTRACT: The hippocampal proteins obtained from both exercised (moderate-intensity running for 6 weeks) and sedentary mice were separately analyzed for Liquid chromatography–tandem mass spectrometry (LC-MS/MS) identification with three replications. Acetylome data identified 3876 acetyl sites and 1764 acetylated proteins, which constituted 31% of the whole hippocampal proteome (totally 5725 proteins identified). In addition, 1305 proteins with 2769 acetyl sites were quantified. When defining the cutoffs for the fold change in abundance of 1.5 and P value < 0.05, 272 acetyl sites on 252 proteins were differentially regulated between exercise and control mice. Analysis of the subcellular localization showed that these differentially acetylated proteins (DAPs) exert different functions in multiple compartments, including the cytoplasm, mitochondria, nucleus and plasma membrane. The 19 up-regulated acetylated proteins mainly resided in mitochondria, while the 253 down-regulated DAPs were most abundant in cytoplasm, and the annotation of molecular functions were significantly enriched in catalytic activity and binding, respectively. Gene Ontology (GO) analysis showed that DAPs were primarily correlated with cytoskeleton, myelin sheath and axons in the cellular component category; and actin cytoskeleton organization, regulation of protein polymerization or depolymerization, and hexose/glucose metabolic process were significantly enriched in the biological process category. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that DAPs were involved in central carbon metabolism, neurodegeneration diseases, synapses, and various signaling pathways. Among them, chronic exercise induced significant alterations in phototransduction, carbon-related metabolism (carbohydrate digestion and absorption, glycolysis/gluconeogenesis, fructose and mannose metabolism, and pyruvate metabolism) and Hippo signaling pathway. Meanwhile, 21 proteins were significantly expressed, which were enriched in the pathway of complement and coagulation cascades.
Shan Wang 
