Proteomic analysis of brain protein expression levels in NF-κβ p50 −/− homozygous knockout mice

Abstract: The role of nuclear factor kappa B (NF-κB) in oxidative stress, and most recently in pro- and anti-apoptotic-related mechanistic pathways, has well been established. Because of the dual nature of NF-κB, the wide range of genes it regulates and the plethora of stimuli that activate it, various studies addressing the functional role of NF-κB proteins have resulted in a number of differing findings. The present study examined the effect of a stimulus-free environment on the frontal cortex of mice brain with the p50 subunit of NF-κB knocked out p50 (−/−). Homozygous p50 mice knockout (KO) and wild type (WT) were used, and at 7–9 weeks they were sacrificed and various brain regions dissected. We analyzed the levels of oxidation in the frontal cortex of both the p50 (−/−) and WT mice. There was a significant reduction in the levels of protein-bound 4-hydroxynonenal (HNE) [a lipid peroxidation product], 3-nitrotyrosine (3NT), and protein carbonyls in the p50 (−/−) mice when compared to the WT. A proteomic profile analysis identified ATP synthase gamma chain, ubiquinol-cyt-C reductase, heat shock protein 10 (Hsp10), fructose bisphosphate aldolase C, and NADH–ubiquinone oxidoreductase as proteins whose expressions were significantly increased in the p50 (−/−) mice compared to the WT. With the reduction in the levels of oxidative stress and the increase in expression of key proteins in the p50 (−/−) brain, this study suggests that the p50 subunit can potentially be targeted for the development of therapeutic interventions in disorders in which oxidative stress plays a key role. [Copyright &y& Elsevier]