Update publication information. The plateau adaptability and stress resistance of yaks is widely known due to their capacity to survive under severe habitat conditions. However, few studies on brain mitochondria have characterized these adaptations at the proteomic level. In the present study, brain mitochondrial proteins of yaks and cattle were identified and quantified using the iTRAQ and proteomics. Western blotting was used to verify changes in abundant of target proteins. A total of 57 DAPs were identified in the yak brain tissue. GO analysis of molecular functions of these proteins revealed reduced oxidoreductase activity, elevated coenzyme binding, downregulated oxidation–reduction processes, and upregulated small molecule metabolic processes. STRING protein interaction analysis indicated a complex interaction between these enzyme families the dehydrogenase family, the transaminase and the ATP synthetase. Reactome pathway analysis highlighted that the majority of differentially abundant proteins participated in aerobic metabolic pathways such as metabolism, the citric acid cycle, and respiratory electron transport. Immunoblotting confirmed that changes in the FKBP4 and ATPAF2 abundant were consistent with the results of mass spectrometry. In the present study, we performed high-throughput screening to identify DAPs in the brain mitochondria of yak and cattle to study the plateau adaptability of yaks.