Abstract

The specificity of age-related oxidative damage to proteomes is largely unknown. Our long-term goal is to develop new proteomic strategies capable of identifying proteins susceptible to such damage. In this exploratory R21 application, our short-term goal is to use the concept of subcellular proteomes, carbonyl immuno-capture, and ICAT technology to identify (and discover) age-related carbonyl-associated proteins in muscle mitochondria. We will use the Fisher 344 rat as an animal model. We will compare the proteomes of two muscles whose functions decline differently with aging, the soleus and semimembranosus. While the soleus is significantly dependent on mitochondria for ATP production, the semimembranosus relies on glycolysis as a source of ATP. As a result, it is hypothesized that different ROS production patterns will cause different protein oxidative damage in these two muscles. Two studies are proposed: (1) direct comparison of carbonyl-associated proteins in the two muscle types at 6 and 28 months of age; (2) a longitudinal study with muscle sampling from the same animal at three different ages. Sample limitations in the second study will be addressed by using FT-ICR-MS and CE-LIF. ? ?

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21AG025371-02
Application #
6950861
Study Section
Special Emphasis Panel (ZAG1-ZIJ-5 (O1))
Program Officer
Kohanski, Ronald A
Project Start
2004-09-30
Project End
2007-07-31
Budget Start
2005-08-01
Budget End
2007-07-31
Support Year
2
Fiscal Year
2005
Total Cost
$148,500
Indirect Cost

  Institution

Name
University of Minnesota Twin Cities
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455

  Publications

Grimsrud, Paul A; Xie, Hongwei; Griffin, Timothy J et al. (2008) Oxidative stress and covalent modification of protein with bioactive aldehydes. J Biol Chem 283:21837-41
Feng, Juan; Xie, Hongwei; Meany, Danni L et al. (2008) Quantitative proteomic profiling of muscle type-dependent and age-dependent protein carbonylation in rat skeletal muscle mitochondria. J Gerontol A Biol Sci Med Sci 63:1137-52
Feng, Juan; Navratil, Marian; Thompson, LaDora V et al. (2008) Estimating relative carbonyl levels in muscle microstructures by fluorescence imaging. Anal Bioanal Chem 391:2591-8
Feng, Juan; Arriaga, Edgar A (2008) Quantification of carbonylated proteins in rat skeletal muscle mitochondria using capillary sieving electrophoresis with laser-induced fluorescence detection. Electrophoresis 29:475-82
Feng, Juan; Navratil, Marian; Thompson, Ladora V et al. (2008) Principal component analysis reveals age-related and muscle-type-related differences in protein carbonyl profiles of muscle mitochondria. J Gerontol A Biol Sci Med Sci 63:1277-88
Grimsrud, Paul A; Picklo Sr, Matthew J; Griffin, Timothy J et al. (2007) Carbonylation of adipose proteins in obesity and insulin resistance: identification of adipocyte fatty acid-binding protein as a cellular target of 4-hydroxynonenal. Mol Cell Proteomics 6:624-37
Roe, Mikel R; Xie, Hongwei; Bandhakavi, Sricharan et al. (2007) Proteomic mapping of 4-hydroxynonenal protein modification sites by solid-phase hydrazide chemistry and mass spectrometry. Anal Chem 79:3747-56
Griffin, Timothy J; Xie, Hongwei; Bandhakavi, Sricharan et al. (2007) iTRAQ reagent-based quantitative proteomic analysis on a linear ion trap mass spectrometer. J Proteome Res 6:4200-9
Meany, Danni L; Xie, Hongwei; Thompson, LaDora V et al. (2007) Identification of carbonylated proteins from enriched rat skeletal muscle mitochondria using affinity chromatography-stable isotope labeling and tandem mass spectrometry. Proteomics 7:1150-63
Griffin, Timothy J; Seth, Gargi; Xie, Hongwei et al. (2007) Advancing mammalian cell culture engineering using genome-scale technologies. Trends Biotechnol 25:401-8

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