The goal of this proposal is to conduct a comprehensive examination of the effect of aging on the mitochondrial proteome of mice. We have chosen to focus on the mitochondrial proteome for two reasons. First, there is almost unanimous agreement that mitochondria are very likely to play an important role in aging, either through changes in function (e.g., reduced energy generation or increase apoptosis) or through the generation of ROS, which in turn damage molecules leading to aging. Second, the mitochondrial proteome has a manageable number of proteins (1000 to 2000) to study using the current technology in proteomics. Although there are substantial data on the deterioration of mitochondrial function with aging, there is little information on the molecular basis of this phenomenon. We hypothesize that alterations in the mitochondrial proteome have a high probability of playing a major role in the age-related changes in mitochondria function. We proposed to test this hypothesis by performing the first detailed study of global expression and post-translational modifications in mitochondria proteome as a function of age by studying mitochondria isolated from various tissues of mice over their life span and determining the effect of dietary restriction (DR), which retards aging, on the age-related changes observed in the mitochondrial proteome.
The Specific Aims of this project are to profile the effects of age and DR on 1) protein expression in the mitochondria proteome, 2) phosphorylation of the mitochondria phosphoproteome, 3) oxidative modification of mitochondria proteins, and 4) mitochondrial function. These studies will be conducted with a series of new proteomics methods we have developed specifically for the examination of protein expression, enzymatic post-translational modifications, and protein oxidation. Recognizing that different proteins are often seen with multidimensional chromatography and gel electrophoresis, we have designed methods that allow us to employ both techniques in our studies of the mitochondrial proteome.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
1R01AG025362-01
Application #
6862551
Study Section
Special Emphasis Panel (ZAG1-ZIJ-5 (O1))
Program Officer
Finkelstein, David B
Project Start
2004-09-30
Project End
2009-07-31
Budget Start
2004-09-30
Budget End
2005-07-31
Support Year
1
Fiscal Year
2004
Total Cost
$254,199
Indirect Cost
Name
Purdue University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Madian, Ashraf G; Hindupur, Jagadish; Hulleman, John D et al. (2012) Effect of single amino acid substitution on oxidative modifications of the Parkinson's disease-related protein, DJ-1. Mol Cell Proteomics 11:M111.010892
Madian, Ashraf G; Diaz-Maldonado, Naomi; Gao, Qiang et al. (2011) Oxidative stress induced carbonylation in human plasma. J Proteomics 74:2395-416
Madian, Ashraf G; Myracle, Angela D; Diaz-Maldonado, Naomi et al. (2011) Differential carbonylation of proteins as a function of in vivo oxidative stress. J Proteome Res 10:3959-72
Madian, Ashraf G; Myracle, Angela D; Diaz-Maldonado, Naomi et al. (2011) Determining the effects of antioxidants on oxidative stress induced carbonylation of proteins. Anal Chem 83:9328-36
Kim, Jin-Hee; Sedlak, Miroslav; Gao, Qiang et al. (2010) Dynamics of protein damage in yeast frataxin mutant exposed to oxidative stress. OMICS 14:689-99
Madian, Ashraf G; Regnier, Fred E (2010) Proteomic identification of carbonylated proteins and their oxidation sites. J Proteome Res 9:3766-80
PĂ©rez, Viviana I; Pierce, Anson; de Waal, Eric M et al. (2010) Detection and quantification of protein disulfides in biological tissues a fluorescence-based proteomic approach. Methods Enzymol 473:161-77
Madian, Ashraf G; Regnier, Fred E (2010) Profiling carbonylated proteins in human plasma. J Proteome Res 9:1330-43
Perez, Viviana I; Buffenstein, Rochelle; Masamsetti, Venkata et al. (2009) Protein stability and resistance to oxidative stress are determinants of longevity in the longest-living rodent, the naked mole-rat. Proc Natl Acad Sci U S A 106:3059-64
Pierce, Anson; Mirzaei, Hamid; Muller, Florian et al. (2008) GAPDH is conformationally and functionally altered in association with oxidative stress in mouse models of amyotrophic lateral sclerosis. J Mol Biol 382:1195-210

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