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Proteome analysis of DNA damage-induced neuronal death using high throughput mass spectrometry

  1. Author:
    Johnson, M. D.
    Yu, L. R.
    Conrads, T. P.
    Kinoshita, Y.
    Uo, T.
    Matthews, J. D.
    Lee, S. W.
    Smith, R. D.
    Veenstra, T. D.
    Morrison, R. S.

  2. Author Address

    Morrison, RS, Univ Washington, Sch Med, Dept Neurol Surg, Box 356470,1959 NE Pacific St, Seattle, WA 98195 USA Univ Washington, Sch Med, Dept Neurol Surg, Seattle, WA 98195 USA. NCI, Biomed Proteom Program, SAIC Frederick, Frederick, MD 21702 USA. Pacific NW Natl Lab, Environm & Mol Sci Lab, Richland, WA 99352 USA. Harvard Univ, Brigham & Womens Hosp, Sch Med, Dept Neurol Surg, Boston, MA 02115 USA.
    1. Year: 2004
  2. Journal: Journal of Biological Chemistry
    1. 279
    2. 25
    3. Pages: 26685-26697
  4. Type of Article: Article
  1. Abstract:

    Isotope-coded affinity tag reagents and high throughput mass spectrometry were used to quantitate changes in the expression of 150 proteins in mouse wild-type (p53(+/+)) cortical neurons undergoing DNA damage-induced death. Immunological techniques confirmed several of the changes in protein expression, but microarray analysis indicated that many of these changes were not accompanied by altered mRNA expression. Proteome analysis revealed perturbations in mitochondrial function, free radical production, and neuritogenesis that were not observed in p53-deficient neurons. Changes in Tau, cofilin, and other proteins recapitulated abnormalities observed in neurodegenerative states in vivo. Additionally, DNA damage caused a p53-dependent decrease in expression of members of the protein kinase A (PKA) signaling pathway. PKA inhibition promoted death in the absence of DNA damage, revealing a novel mechanism by which endogenous down-regulation of PKA signaling may contribute to p53-dependent neuronal death. These data demonstrate the power of high throughput mass spectrometry for quantitative analysis of the neuronal proteome

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