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High Fat Diet Related Changes in Mitochondrial Proteome

ABSTRACT: Western diets leads to obesity and its comorbidities, in particular the progression of hepatosteatosis and insulin resistance. In DIO C57/BL6 obese mice, we performed quantitative mitochondrial bioenergetics and proteomic analysis and fused both technologies to understand the functional adjustments of mitochondrial energy metabolism in response to overfeeding.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver

SUBMITTER: Jacek Wisniewski

LAB HEAD: Jacek R. Wisniewski

PROVIDER: PXD002289 | Pride | 2018-02-20

REPOSITORIES: Pride

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			Action	DRS
	20141119_Qep7_JRW_SA_G21L.raw	Raw
	20141119_Qep7_JRW_SA_G21T.raw	Raw
	20141119_Qep7_JRW_SA_G22L.raw	Raw
	20141119_Qep7_JRW_SA_G22T.raw	Raw
	20141119_Qep7_JRW_SA_L_G25.raw	Raw

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Publications

Respiromics - An integrative analysis linking mitochondrial bioenergetics to molecular signatures.

Walheim Ellen E Wiśniewski Jacek R JR Jastroch Martin M

Molecular metabolism 20180105

<h4>Objective</h4>Energy metabolism is challenged upon nutrient stress, eventually leading to a variety of metabolic diseases that represent a major global health burden.<h4>Methods</h4>Here, we combine quantitative mitochondrial respirometry (Seahorse technology) and proteomics (LC-MS/MS-based total protein approach) to understand how molecular changes translate to changes in mitochondrial energy transduction during diet-induced obesity (DIO) in the liver.<h4>Results</h4>The integrative analysi ...[more]

PMID: 29361498

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Project description:Glycolysis is the core metabolic pathway supplying energy to cells. Whereas the vast majority of studies focus on specific aspects of the process, global analyses characterizing simultaneously all enzymes involved in the process are scarce. Here we demonstrate that quantitative label- and standard-free proteomics allows accurate determination of titers of metabolic enzymes and enables simultaneous measurements of titers and maximal enzymatic activities (Amax)c of all glycolytic enzymes and the gluconeogenic fructose 1,6-bisphosphatase in mouse brain, liver and muscle. Despite occurrence of tissue-specific isoenzymes bearing different kinetic properties, the enzyme titers often correlated well with the Amax values. To provide a more general picture of energy metabolism we analyzed titers of the enzymes in additional 7 mouse organs and in human cells. Across the analyzed samples we identified two basic profiles: a 'fast glucose uptake' one in brain and heart, and a 'gluconeogenic rich' one occurring in liver. In skeletal muscles and other organs, we found intermediate profiles. Obtained data highlighted the glucose-flux-limiting role of hexokinase which activity was always 10-100-fold lower than the average activity of all other glycolytic enzymes. A parallel determination of enzyme titers and maximal enzymatic activities allowed determination of kcat values without enzyme purification. Results of our in-depth proteomic analysis of the mouse organs did not support the concepts of regulation of glycolysis by lysine acetylation.

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Proteomic and phosphoproteomic characterization of steatotic liver from diet-induced obese mice in response to dietary fat

Project description:One of the metabolic consequences of obesity is abnormal lipid accumulation in the liver, or hepatosteatosis, which can develop intomore serious diseases in the non-alcoholic liver disease (NAFLD) spectrum including non-alcoholic steatohepatitis (NASH) and hepatocellular carcinoma. Therefore, it is necessary to understand the metabolic changes that occur in hepatosteatosisin order toprevent disease progression.The liver plays a major role in regulating macronutrient and energy homeostasis in both the fed and fasted states. However, whether hepatosteatosis influences thepostprandial molecularresponse, specifically to dietary fat,has not been investigated.Therefore, the goal of this study was to comparethe proteome and phosphoproteome of steatotic livers from diet-induced obese (DIO) mice and control livers from lean mice in the postprandial response to dietary fat. Using untargeted LC-MS/MS analysis, we identified significant alterations in the levels of proteins involved in macronutrient and energy metabolism in livers of DIO compared to lean mice. In addition, uniquely phosphorylated proteins in livers of DIO andlean mice reflect regulatory mechanisms controlling cellular processes contributingto hepatosteatosis. Theresultsof this studyexpandourknowledge ofthe metabolic consequences that occur duringhepatosteatosisandtheinfluence of dietary fat on NAFLD progression.

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Irp2 mediates cigarette smoke-induced bronchitis and emphysema via regulation of cytochrome c oxidase and mitochondrial iron loading.

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