The Story Akin To AZD2014
To do so, we performed these analyses in two phases. In the univariate phase, we analyzed liver mitochondria from four lean (wild-type) and four obese (leptin?/?, ob/ob) 10-week-old B6 mice ( Figure?1A). This initial phase enabled us to assess the breadth and depth at which our analysis platform could detect changes to the mitochondrial proteome selleck and phosphoproteome. In the multivariate phase, we analyzed 40 mice differing by three variables: age, strain, and obesity ( Figure?1A). These animals included lean (wild-type) and obese (ob/ob) mice from both diabetes-susceptible (BTBR) and diabetes-resistant (B6) strains, each as either adolescents (4?weeks of age) or adults (10?weeks of age). For each phase, we purified mitochondria only to the extent required to achieve near-comprehensive coverage of the liver MitoCarta protein list, allowing us to simultaneously profile copurifying organelles (MitoCarta is a tissue-specific compendium of mitochondrial proteins complied using MS-based proteomic analyses of highly purified mitochondria, machine learning, and GFP microscopy [Pagliarini et?al., 2008]). Our univariate phase identified 3,447 unique proteins and 3,895 unique phosphoisoforms (site-specific phosphorylation patterns) from just over 1 million MS/MS scans. These include 692 of selleck chemical the 700 liver MitoCarta proteins, and 449 mitochondrial phosphoisoforms. Notably, unsupervised hierarchical clustering of the protein abundance measurements appropriately grouped the mice by?their obesity status (Figure?1B). After correcting for multiple hypotheses, significant differences (q?OPHN1 for 1,014 proteins (325 mitochondrial, Figures 1H and see Figure?S1B online) and 720 phosphoisoforms (102 mitochondrial, Figure?1H and Figure?S1D). Our univariate data revealed extensive and reproducible remodeling of the liver mitochondrial proteome and phosphoproteome with obesity (Figures 1B�C1D, Figures S1B�CS1D). For example, fatty acid oxidation and oxidative phosphorylation (OxPhos) proteins were increased with obesity (Figures 1C and 1G), while reactive oxygen species (ROS) detoxification enzymes were decreased (Figure?S1C), consistent with known liver alterations (Buchner et?al., 2011; Deng et?al., 2010; Wang et?al., 2012a). Phosphorylation changes were also prominent in?a range?of central mitochondrial (and nonmitochondrial) pathways (see http://mitomod.biochem.wisc.edu and Table S1, Table S2, Table S3, Table S4, and Table S5). For example, the three most statistically significant changes (q?
